Optimizing Biocompatibility and Gene Delivery with DMAEA and DMAEAm: A Niacin-Derived Copolymer ApproachClick to copy article linkArticle link copied!
- Prosper P. MapfumoProsper P. MapfumoInstitute of Organic and Macromolecular Chemistry (IOMC), Friedrich Schiller University Jena, Humboldtstrasse 10, Jena 07743, GermanyMore by Prosper P. Mapfumo
- Liên S. ReichelLiên S. ReichelInstitute of Organic and Macromolecular Chemistry (IOMC), Friedrich Schiller University Jena, Humboldtstrasse 10, Jena 07743, GermanyMore by Liên S. Reichel
- Thomas AndréThomas AndréLeibniz Institute on Aging-Fritz Lipmann Institute, Jena 07745, GermanyMore by Thomas André
- Stephanie HoeppenerStephanie HoeppenerInstitute of Organic and Macromolecular Chemistry (IOMC), Friedrich Schiller University Jena, Humboldtstrasse 10, Jena 07743, GermanyJena Center for Soft Matter (JCSM), Friedrich Schiller University Jena, Philosophenweg 7, Jena 07743, GermanyMore by Stephanie Hoeppener
- Lenhard K. RudolphLenhard K. RudolphLeibniz Institute on Aging-Fritz Lipmann Institute, Jena 07745, GermanyMore by Lenhard K. Rudolph
- Anja Traeger*Anja Traeger*Email: [email protected]Institute of Organic and Macromolecular Chemistry (IOMC), Friedrich Schiller University Jena, Humboldtstrasse 10, Jena 07743, GermanyJena Center for Soft Matter (JCSM), Friedrich Schiller University Jena, Philosophenweg 7, Jena 07743, GermanyMore by Anja Traeger
Abstract
Gene therapy is pivotal in nanomedicine, offering a versatile approach to disease treatment. This study aims to achieve an optimal balance between biocompatibility and efficacy, which is a common challenge in the field. A copolymer library is synthesized, incorporating niacin-derived monomers 2-acrylamidoethyl nicotinate (AAEN) or 2-(acryloyloxy)ethyl nicotinate (AEN) with N,N-(dimethylamino)ethyl acrylamide (DMAEAm) or hydrolysis-labile N,N-(dimethylamino)ethyl acrylate (DMAEA). Evaluation of the polymers’ cytotoxicity profiles reveals that an increase in AAEN or DMAEA molar ratios correlates with improved biocompatibility. Remarkably, an increase in AAEN in both DMAEA and DMAEAm copolymers demonstrated enhanced transfection efficiencies of plasmid DNA in HEK293T cells. Additionally, the top-performing polymers demonstrate promising gene expression in challenging-to-transfect cells (THP-1 and Jurkat cells) and show no significant effect on modulating immune response induction in ex vivo treated murine monocytes. Overall, the best performing candidates exhibit an optimal balance between biocompatibility and efficacy, showcasing potential advancements in gene therapy.
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License Summary*
You are free to share(copy and redistribute) this article in any medium or format and to adapt(remix, transform, and build upon) the material for any purpose, even commercially within the parameters below:
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*Disclaimer
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License Summary*
You are free to share(copy and redistribute) this article in any medium or format and to adapt(remix, transform, and build upon) the material for any purpose, even commercially within the parameters below:
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Attribution (BY): Credit must be given to the creator.
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Introduction
Experimental Section
Polymer Synthesis
Synthesis of P(AAEN148/123/48-co-DMAEAm50/75/142) (A1–A3)
Synthesis of P(AAEN152/120/49-co-DMAEA50/72/149) (B1–B3)
Synthesis of P(AEN115-co-DMAEA65) (C1)
Titrations
DMAEA Copolymer Degradation
Biological Assays
Cell Culture
Cytocompatibility (PrestoBlue Assay)
Polymer–Membrane Interaction
Polyplexation
Size Determination via Dynamic Light Scattering
Interaction of Polymers and Genetic Material
Cryogenic Transmission Electron Microscopy (Cryo-TEM)
Particle Uptake
Transfection Efficiency
Cytocompatibility of Polyplexes (CytoTox-ONE Assay)
Endosomal Release
Ex Vivo Monocyte Culture
RNA Isolation and RT-qPCR
Statistical Analysis
Results and Discussion
Synthesis and Characterization
Figure 1
Figure 1. (A) Reaction scheme illustrates the structures of the polymer library and their polymerization conditions. (B) SEC traces of the polymer library using DMAc (0.21 wt % LiCl) as eluent. (C) Titration curves of the polymer library were determined using 0.15 M NaOH as the base with a Metrohm OMNIS integrated titration system.
A1 | A2 | A3 | B1 | B2 | B3 | C1 | |
---|---|---|---|---|---|---|---|
Mn,tha (kg mol–1) | 40.1 | 38.0 | 31.1 | 40.8 | 37.0 | 32.3 | 34.9 |
Mn,SECb, (kg mol–1) | 41.0 | 41.5 | 33.2 | 40.5 | 36.8 | 26.5 | 14.7 |
D̵b | 1.3 | 1.3 | 1.3 | 1.3 | 1.3 | 1.3 | 1.4 |
pKacDMAEAm/DMAEA | 7.6 | 7.8 | 8.0 | 7.2 | 7.5 | 7.7 | 7.3 |
Calculated via conversion using 1H NMR and eq S1.
Determined by SEC using DMAc (0.21 wt % LiCl) as eluent and PMMA standards for calibration.
Polyplex Formation and Characterization
Figure 2
Figure 2. (A) Experiment design of EBA (first two steps) and HRA (all three steps). (B) EBA was performed at different N*/P ratios ranging from 3 to 20. (C) HRA assay at N*/P = 20. All data points were performed in triplicate, and values were fitted using a B-Spline function. (D) Hydrodynamic size measurement via DLS. (E) Cryo-TEM image of polyplexes of C1.
Cytotoxicity and Polymer–Membrane Interaction
Figure 3
Figure 3. (A) Mechanism of the cytotoxicity assay (PrestoBlue assay). (B) PrestoBlue assay in L929 over 24 h in a full growth medium (D10H). Dots represent values of single repetitions. Lines were fitted, and IC50 values were calculated with dose–response function (n = 3). Stars indicate the polymer concentration (μg mL–1), which induces 50% cytotoxicity. Viability below 70% was considered cytotoxic. (C) Hemolysis assay was performed in triplicate with three different donors. A relative hemolysis of > 2% is considered slightly hemolytic, and > 5% is considered hemolytic.
Particle Uptake Study
Figure 4
Figure 4. (A) Schematic illustration of complexation between the polymers and pDNA and YOYO-1. The experiment was used to determine the particle uptake behavior. (B) Particle uptake was performed in HEK293T cells with full growth medium (D10H) over 1 and 4 h at N*/P ratio 20 and c(pDNA) = 3 μg mL–1 on cells (n = 3). The gating strategy can be found in the Supporting Information, Figure S9.
Transfection Efficiency
Figure 5
Figure 5. (A) Schematic illustration of the experimental design for determining transfection efficiency under two different conditions (4 + 20 and 24 h). (B) Transfection efficiency was performed in HEK293T cells with full growth medium (D10H) over 4 + 20 h at N*/P 20 and c(pDNA) = 3 μg mL–1 on cells (n = 3). (C) Performed over 24 h at N*/P 20 and three different concentrations of the genetic material (n = 3). Details of statistical tests can be found in the Supporting Information, Tables S6–S11.
Figure 6
Figure 6. Transfection efficiency was performed in HEK293T cells with full growth medium (D10H) over 24 h at different N*/P ratios and c(pDNA) = 3 μg mL–1 on cells for A1, B2, and C1 (n ≥ 3). Details of statistical tests can be found in the Supporting Information, Tables S12–S15.
Immunomodulatory Role of Niacin-Derived Copolymer
Figure 7
Figure 7. (A) Transfection efficiency was performed with A1, B1, B2, and C1 in hard-to-transfect suspension cell lines THP-1 and Jurkat. Transfection was performed in full growth medium (R10H) over 24 h at N*/P 20 and 3 μg mL–1 genetic material on cells (n = 3). (B) Fold change (compared to non-treated cells) of the relative TNF-α mRNA expression (normalized to GAPDH) of ex vivo murine classical monocytes from the bone marrow. One point represents one biological replicate (mice, blue = male, black = female). Mean with standard deviation is depicted. Details of statistical tests can be found in the Supporting Information, Tables S16 and S17.
Conclusions
Supporting Information
The Supporting Information is available free of charge at https://pubs.acs.org/doi/10.1021/acs.biomac.4c00007.
Instruments and materials, monomer and polymer synthesis and characterization, polymerization kinetics procedures, titrations, degradation of DMAEA, N*/P ratio calculations, EBA and HRA, cytocompatibility (PrestoBlue and CytoTox-One assay), particle uptake study, transfection efficiency, endosomal escape, antibody mix for murine monocyte staining, and statistics (PDF)
Terms & Conditions
Most electronic Supporting Information files are available without a subscription to ACS Web Editions. Such files may be downloaded by article for research use (if there is a public use license linked to the relevant article, that license may permit other uses). Permission may be obtained from ACS for other uses through requests via the RightsLink permission system: http://pubs.acs.org/page/copyright/permissions.html.
Acknowledgments
The authors thankfully acknowledge Carolin Kellner and Sandra Henk for performing toxicity assays and EBA/HRA assay, taking care of the cell culture, and pDNA preparation and Elisabeth Moek for her support in transfection efficiency assay. Furthermore, we acknowledge Prof. U.S. Schubert for providing excellent facilities. Figures are partially created with BioRender.com.
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- 6Sayed, N.; Allawadhi, P.; Khurana, A.; Singh, V.; Navik, U.; Pasumarthi, S. K.; Khurana, I.; Banothu, A. K.; Weiskirchen, R.; Bharani, K. K. Gene therapy: Comprehensive overview and therapeutic applications. Life Sci. 2022, 294, 120375, DOI: 10.1016/j.lfs.2022.120375Google Scholar6https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BB38XivF2iu74%253D&md5=f76332487e207cdcdeecc77113c5a072Gene therapy: Comprehensive overview and therapeutic applicationsSayed, Nilofer; Allawadhi, Prince; Khurana, Amit; Singh, Vishakha; Navik, Umashanker; Pasumarthi, Sravan Kumar; Khurana, Isha; Banothu, Anil Kumar; Weiskirchen, Ralf; Bharani, Kala KumarLife Sciences (2022), 294 (), 120375CODEN: LIFSAK; ISSN:0024-3205. (Elsevier B.V.)A review. Gene therapy is the product of man's quest to eliminate diseases. Gene therapy has three facets namely, gene silencing using siRNA, shRNA and miRNA, gene replacement where the desired gene in the form of plasmids and viral vectors, are directly administered and finally gene editing based therapy where mutations are modified using specific nucleases such as zinc-finger nucleases (ZFNs), transcription activator-like effector nucleases (TALENs) and clustered regulatory interspaced short tandem repeats (CRISPR)/CRISPR-assocd. protein (Cas)-assocd. nucleases. Transfer of gene is either through transformation where under specific conditions the gene is directly taken up by the bacterial cells, transduction where a bacteriophage is used to transfer the genetic material and lastly transfection that involves forceful delivery of gene using either viral or non-viral vectors. The non-viral transfection methods are subdivided into phys., chem. and biol. The phys. methods include electroporation, biolistic, microinjection, laser, elevated temp., ultrasound and hydrodynamic gene transfer. The chem. methods utilize calcium- phosphate, DAE-dextran, liposomes and nanoparticles for transfection. The biol. methods are increasingly using viruses for gene transfer, these viruses could either integrate within the genome of the host cell conferring a stable gene expression, whereas few other non-integrating viruses are episomal and their expression is dild. proportional to the cell division. So far, gene therapy has been wielded in a plethora of diseases. However, coherent and innocuous delivery of genes is among the major hurdles in the use of this promising therapy. Hence this review aims to highlight the current options available for gene transfer along with the advantages and limitations of every method.
- 7Gaspar, R.; Coelho, F.; Silva, B. F. Lipid-nucleic acid complexes: Physicochemical aspects and prospects for cancer treatment. Molecules 2020, 25 (21), 5006, DOI: 10.3390/molecules25215006Google Scholar7https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BB3cXitleit7jJ&md5=a3870e755b37ace60de4b9ed943abc4aLipid-nucleic acid complexes: physicochemical aspects and prospects for cancer treatmentGaspar, Ricardo; Coelho, Filipe; Silva, Bruno F. B.Molecules (2020), 25 (21), 5006CODEN: MOLEFW; ISSN:1420-3049. (MDPI AG)A review. Cancer is an extremely complex disease, typically caused by mutations in cancer-crit. genes. By delivering therapeutic nucleic acids (NAs) to patients, gene therapy offers the possibility to supplement, repair or silence such faulty genes or to stimulate their immune system to fight the disease. While the challenges of gene therapy for cancer are significant, the latter approach (a type of immunotherapy) starts showing promising results in early-stage clin. trials. One important advantage of NA-based cancer therapies over synthetic drugs and protein treatments is the prospect of a more universal approach to designing therapies. Designing NAs with different sequences, for different targets, can be achieved by using the same technologies. This versatility and scalability of NA drug design and prodn. on demand open the way for more efficient, affordable and personalized cancer treatments in the future. However, the delivery of exogenous therapeutic NAs into the patients' targeted cells is also challenging. Membrane-type lipids exhibiting permanent or transient cationic character have been shown to assoc. with NAs (anionic), forming nanosized lipid-NA complexes. These complexes form a wide variety of nanostructures, depending on the global formulation compn. and properties of the lipids and NAs. Importantly, these different lipid-NA nanostructures interact with cells via different mechanisms and their therapeutic potential can be optimized to promising levels in vitro. The complexes are also highly customizable in terms of surface charge and functionalization to allow a wide range of targeting and smart-release properties. Most importantly, these synthetic particles offer possibilities for scaling-up and affordability for the population at large. Hence, the versatility and scalability of these particles seem ideal to accommodate the versatility that NA therapies offer. While in vivo efficiency of lipid-NA complexes is still poor in most cases, the advances achieved in the last three decades are significant and very recently a lipid-based gene therapy medicine was approved for the first time (for treatment of hereditary transthyretin amyloidosis). Although the path to achieve efficient NA-delivery in cancer therapy is still long and tenuous, these advances set a new hope for more treatments in the future. In this review, we attempt to cover the most important biophys. and physicochem. aspects of non-viral lipid-based gene therapy formulations, with a perspective on future cancer treatments in mind.
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- 10Shi, B.; Zheng, M.; Tao, W.; Chung, R.; Jin, D.; Ghaffari, D.; Farokhzad, O. C. Challenges in DNA delivery and recent advances in multifunctional polymeric DNA delivery systems. Biomacromolecules 2017, 18 (8), 2231– 2246, DOI: 10.1021/acs.biomac.7b00803Google Scholar10https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC2sXhtVOnsbnL&md5=bff2ce2f4f341f3b182f4da5d3840a61Challenges in DNA Delivery and Recent Advances in a Multifunctional Polymeric DNA Delivery SystemsShi, Bingyang; Zheng, Meng; Tao, Wei; Chung, Roger; Jin, Dayong; Ghaffari, Dariush; Farokhzad, Omid C.Biomacromolecules (2017), 18 (8), 2231-2246CODEN: BOMAF6; ISSN:1525-7797. (American Chemical Society)After more than 20 years of intensive investigations, gene therapy has become one of the most promising strategies for treating genetic diseases. However, the lack of ideal delivery systems has limited the clin. realization of gene therapy's tremendous potential, esp. for DNA-based gene therapy. Over the past decade, considerable advances have been made in the application of polymer-based DNA delivery systems for gene therapy, esp. through multifunctional systems. The core concept behind multifunctional polymeric DNA delivery systems is to endow one single DNA carrier, via materials engineering and surface modification, with several active functions, e.g., good cargo DNA protection, excellent colloidal stability, high cellular uptake efficiency, efficient endo/lysosome escape, effective import into the nucleus, and DNA unpacking. Such specially developed vectors would be capable of overcoming multiple barriers to the successful delivery of DNA. In this review, we first provide a comprehensive overview of the interactions between the protein corona and DNA vectors, the mechanisms and challenges of nonviral DNA vectors, and important concepts in the design of DNA carriers identified via past reports on DNA delivery systems. Finally, we highlight and discuss recent advances in multifunctional polymeric DNA delivery systems based on "off-the-shelf" polycations including polyethylenimine (PEI), poly-L-lysine (PLL), and chitosan and offer perspectives on future developments.
- 11Guo, X.; Huang, L. Recent advances in nonviral vectors for gene delivery. Acc. Chem. Res. 2012, 45 (7), 971– 979, DOI: 10.1021/ar200151mGoogle Scholar11https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC3MXhtVylsbvK&md5=5b429971bf547460e451ad362d6edcaeRecent Advances in Nonviral Vectors for Gene DeliveryGuo, Xia; Huang, LeafAccounts of Chemical Research (2012), 45 (7), 971-979CODEN: ACHRE4; ISSN:0001-4842. (American Chemical Society)A review. Gene therapy has long been regarded a promising treatment for many diseases, whether acquired (such as AIDS or cancer) or inherited through a genetic disorder. A drug based on a nucleic acid, however, must be delivered to the interior of the target cell while surviving an array of biol. defenses honed by evolution. Successful gene therapy is thus dependent on the development of an efficient delivery vector. Researchers have pursued two major vehicles for gene delivery: viral and nonviral (synthetic) vectors. Although viral vectors currently offer greater efficiency, nonviral vectors, which are typically based on cationic lipids or polymers, are preferred because of safety concerns with viral vectors. So far, nonviral vectors can readily transfect cells in culture, but efficient nanomedicines remain far removed from the clinic. Overcoming the obstacles assocd. with nonviral vectors to improve the delivery efficiency and therapeutic effect of nucleic acids is thus an active area of current research. The difficulties are manifold, including the strong interaction of cationic delivery vehicles with blood components, uptake by the reticuloendothelial system (RES), toxicity, and managing the targeting ability of the carriers with respect to the cells of interest. Modifying the surface with poly(ethylene glycol), i.e., PEGylation, is the predominant method used to reduce the binding of plasma proteins to nonviral vectors and minimize clearance by the RES after i.v. administration. Nanoparticles that are not rapidly cleared from the circulation accumulate in the tumors because of the enhanced permeability and retention effect, and the targeting ligands attached to the distal end of the PEGylated components allow binding to the receptors on the target cell surface. Neutral and anionic liposomes have been also developed for systemic delivery of nucleic acids in exptl. animal models. Other approaches include (i) designing and synthesizing novel cationic lipids and polymers, (ii) chem. coupling the nucleic acid to peptides, targeting ligands, polymers, or environmentally sensitive moieties, and (iii) utilizing inorg. nanoparticles in nucleic acid delivery. Recently, the different classes of nonviral vectors appear to be converging, and the ability to combine features of different classes of nonviral vectors in a single strategy has emerged. With the strengths of several approaches working in concert, more hurdles assocd. with efficient nucleic acid delivery might therefore be overcome. In this Account, we focus on these novel nonviral vectors, which are classified as multifunctional hybrid nucleic acid vectors, novel membrane/core nanoparticles for nucleic acid delivery, and ultrasound-responsive nucleic acid vectors. We highlight systemic delivery studies and consider the future prospects for nucleic acid delivery. A better understanding of the fate of the nanoparticles inside the cell and of the interactions between the parts of hybrid particles should lead to a delivery system suitable for clin. use. We also underscore the value of sustained release of a nucleic acid in this endeavor; making vectors targeted to cells with sustained release in vivo should provide an interesting research challenge.
- 12Hou, X.; Zaks, T.; Langer, R.; Dong, Y. Lipid nanoparticles for mRNA delivery. Nat. Rev. Mater. 2021, 6 (12), 1078– 1094, DOI: 10.1038/s41578-021-00358-0Google Scholar12https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BB3MXhslOnsr3N&md5=651ec4d4569c58d0ad34c9215c02fd38Lipid nanoparticles for mRNA deliveryHou, Xucheng; Zaks, Tal; Langer, Robert; Dong, YizhouNature Reviews Materials (2021), 6 (12), 1078-1094CODEN: NRMADL; ISSN:2058-8437. (Nature Portfolio)A review. MRNA (mRNA) has emerged as a new category of therapeutic agent to prevent and treat various diseases. To function in vivo, mRNA requires safe, effective and stable delivery systems that protect the nucleic acid from degrdn. and that allow cellular uptake and mRNA release. Lipid nanoparticles have successfully entered the clinic for the delivery of mRNA; in particular, lipid nanoparticle-mRNA vaccines are now in clin. use against coronavirus disease 2019 (COVID-19), which marks a milestone for mRNA therapeutics. In this Review, we discuss the design of lipid nanoparticles for mRNA delivery and examine physiol. barriers and possible administration routes for lipid nanoparticle-mRNA systems. We then consider key points for the clin. translation of lipid nanoparticle-mRNA formulations, including good manufg. practice, stability, storage and safety, and highlight preclin. and clin. studies of lipid nanoparticle-mRNA therapeutics for infectious diseases, cancer and genetic disorders. Finally, we give an outlook to future possibilities and remaining challenges for this promising technol.
- 13Kumar, R.; Santa Chalarca, C. F.; Bockman, M. R.; Bruggen, C. V.; Grimme, C. J.; Dalal, R. J.; Hanson, M. G.; Hexum, J. K.; Reineke, T. M. Polymeric delivery of therapeutic nucleic acids. Chem. Rev. 2021, 121 (18), 11527– 11652, DOI: 10.1021/acs.chemrev.0c00997Google Scholar13https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BB3MXpvFKgt78%253D&md5=4f4c596405b997bb94c2973d994547e2Polymeric Delivery of Therapeutic Nucleic AcidsKumar, Ramya; Santa Chalarca, Cristiam F.; Bockman, Matthew R.; Bruggen, Craig Van; Grimme, Christian J.; Dalal, Rishad J.; Hanson, Mckenna G.; Hexum, Joseph K.; Reineke, Theresa M.Chemical Reviews (Washington, DC, United States) (2021), 121 (18), 11527-11652CODEN: CHREAY; ISSN:0009-2665. (American Chemical Society)A review. The advent of genome editing has transformed the therapeutic landscape for several debilitating diseases, and the clin. outlook for gene therapeutics has never been more promising. The therapeutic potential of nucleic acids has been limited by a reliance on engineered viral vectors for delivery. Chem. defined polymers can remediate technol., regulatory, and clin. challenges assocd. with viral modes of gene delivery. Because of their scalability, versatility, and exquisite tunability, polymers are ideal biomaterial platforms for delivering nucleic acid payloads efficiently while minimizing immune response and cellular toxicity. While polymeric gene delivery has progressed significantly in the past four decades, clin. translation of polymeric vehicles faces several formidable challenges. The aim of our Account is to illustrate diverse concepts in designing polymeric vectors towards meeting therapeutic goals of in vivo and ex vivo gene therapy. Here, we highlight several classes of polymers employed in gene delivery and summarize the recent work on understanding the contributions of chem. and architectural design parameters. We touch upon characterization methods used to visualize and understand events transpiring at the interfaces between polymer, nucleic acids, and the physiol. environment. We conclude that interdisciplinary approaches and methodologies motivated by fundamental questions are key to designing high-performing polymeric vehicles for gene therapy.
- 14Pack, D. W.; Hoffman, A. S.; Pun, S.; Stayton, P. S. Design and development of polymers for gene delivery. Nat. Rev. Drug Discovery 2005, 4 (7), 581– 593, DOI: 10.1038/nrd1775Google Scholar14https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BD2MXmsFGjtbo%253D&md5=ed3dcd52a8b297ee3d0c1cbb7d5ae43bDesign and development of polymers for gene deliveryPack, Daniel W.; Hoffman, Allan S.; Pun, Suzie; Stayton, Patrick S.Nature Reviews Drug Discovery (2005), 4 (7), 581-593CODEN: NRDDAG; ISSN:1474-1776. (Nature Publishing Group)A review. The lack of safe and efficient gene-delivery methods is a limiting obstacle to human gene therapy. Synthetic gene-delivery agents, although safer than recombinant viruses, generally do not possess the required efficacy. In recent years, a variety of effective polymers have been designed specifically for gene delivery, and much has been learned about their structure-function relationships. With the growing understanding of polymer gene-delivery mechanisms and continued efforts of creative polymer chemists, it is likely that polymer-based gene-delivery systems will become an important tool for human gene therapy.
- 15Durymanov, M.; Reineke, J. Non-viral Delivery of Nucleic Acids: Insight Into Mechanisms of Overcoming Intracellular Barriers. Front. Pharmacol 2018, 9, 971, DOI: 10.3389/fphar.2018.00971Google Scholar15https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC1MXjtVartLw%253D&md5=99126dd3217118f202bc8516198b6b79Non-viral delivery of nucleic acids: insight into mechanisms of overcoming intracellular barriersDurymanov, Mikhail; Reineke, JoshuaFrontiers in Pharmacology (2018), 9 (), 971/1-971/15CODEN: FPRHAU; ISSN:1663-9812. (Frontiers Media S.A.)A review. Delivery of genes, including plasmid DNAs, short interfering RNAs (siRNAs), and mRNAs (mRNAs), using artificial non-viral nanotherapeutics is a promising approach in cancer gene therapy. However, multiple physiol. barriers upon systemic administration remain a key challenge in clin. translation of anti-cancer gene therapeutics. Besides extracellular barriers including sequestration of gene delivery nanoparticles from the bloodstream by resident organ-specific macrophages, and their poor extravasation and tissue penetration in tumors, overcoming intracellular barriers is also necessary for successful delivery of nucleic acids. Whereas for RNA delivery the endosomal barrier holds a key importance, transfer of DNA cargo addnl. requires translocation into the nucleus. Better understanding of crossing membrane barriers by nucleic acid nanoformulations is essential to the improvement of current non-viral carriers. This review aims to summarize relevant literature on intracellular trafficking of non-viral nanoparticles and det. key factors toward surmounting intracellular barriers. Moreover, recent data allowed us to propose new interpretations of current hypotheses of endosomal escape mechanisms of nucleic acid nanoformulations.
- 16Jhaveri, A. M.; Torchilin, V. P. Multifunctional polymeric micelles for delivery of drugs and siRNA. Front. Pharmacol 2014, 5, 77, DOI: 10.3389/fphar.2014.00077Google Scholar16https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A280%3ADC%252BC2cflvVKisA%253D%253D&md5=faf83465811984713e2bc64c8e358819Multifunctional polymeric micelles for delivery of drugs and siRNAJhaveri Aditi M; Torchilin Vladimir PFrontiers in pharmacology (2014), 5 (), 77 ISSN:1663-9812.Polymeric micelles, self-assembling nano-constructs of amphiphilic copolymers with a core-shell structure have been used as versatile carriers for delivery of drugs as well as nucleic acids. They have gained immense popularity owing to a host of favorable properties including their capacity to effectively solubilize a variety of poorly soluble pharmaceutical agents, biocompatibility, longevity, high stability in vitro and in vivo and the ability to accumulate in pathological areas with compromised vasculature. Moreover, additional functions can be imparted to these micelles by engineering their surface with various ligands and cell-penetrating moieties to allow for specific targeting and intracellular accumulation, respectively, to load them with contrast agents to confer imaging capabilities, and incorporating stimuli-sensitive groups that allow drug release in response to small changes in the environment. Recently, there has been an increasing trend toward designing polymeric micelles which integrate a number of the above functions into a single carrier to give rise to "smart," multifunctional polymeric micelles. Such multifunctional micelles can be envisaged as key to improving the efficacy of current treatments which have seen a steady increase not only in hydrophobic small molecules, but also in biologics including therapeutic genes, antibodies and small interfering RNA (siRNA). The purpose of this review is to highlight recent advances in the development of multifunctional polymeric micelles specifically for delivery of drugs and siRNA. In spite of the tremendous potential of siRNA, its translation into clinics has been a significant challenge because of physiological barriers to its effective delivery and the lack of safe, effective and clinically suitable vehicles. To that end, we also discuss the potential and suitability of multifunctional polymeric micelles, including lipid-based micelles, as promising vehicles for both siRNA and drugs.
- 17Gardlík, R.; Pálffy, R.; Hodosy, J.; Lukács, J.; Turna, J.; Celec, P. Vectors and delivery systems in gene therapy. Med. Sci. Monit. 2005, 11 (4), RA110Google ScholarThere is no corresponding record for this reference.
- 18Perrier, S. 50th Anniversary Perspective: RAFT Polymerization─ A User Guide. Macromolecules 2017, 50 (19), 7433– 7447, DOI: 10.1021/acs.macromol.7b00767Google Scholar18https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC2sXhsFGms7vL&md5=e58928228fbb93e6392106d4abce2a4a50th Anniversary Perspective: RAFT Polymerization-A User GuidePerrier, SebastienMacromolecules (Washington, DC, United States) (2017), 50 (19), 7433-7447CODEN: MAMOBX; ISSN:0024-9297. (American Chemical Society)A review. This Perspective summarizes the features and limitations of reversible addn.-fragmentation chain transfer (RAFT) polymn., highlighting its strengths and weaknesses, as our understanding of the process, from both a mechanistic and an application point of view, has matured over the past 20 years. It is aimed at both experts in the field and newcomers, including undergraduate and postgraduate students, as well as nonexperts in polymn. who are interested in developing their own polymeric structures by exploiting the simple setup of a RAFT polymn.
- 19Gregory, A.; Stenzel, M. H. Complex polymer architectures via RAFT polymerization: From fundamental process to extending the scope using click chemistry and nature’s building blocks. Prog. Polym. Sci. 2012, 37 (1), 38– 105, DOI: 10.1016/j.progpolymsci.2011.08.004Google Scholar19https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC3MXhsVaqt7%252FO&md5=43cfa17e622a10cf00c31388f74d06d0Complex polymer architectures via RAFT polymerization: From fundamental process to extending the scope using click chemistry and nature's building blocksGregory, Andrew; Stenzel, Martina H.Progress in Polymer Science (2012), 37 (1), 38-105CODEN: PRPSB8; ISSN:0079-6700. (Elsevier Ltd.)A review. Reversible addn. fragmentation chain transfer (RAFT) polymn. has made a huge impact in macromol. design. The first block copolymers were described early on, followed by star polymers and then graft polymers. In the last five years, the types of architectures available have become more and more complex. Star and graft polymers now have block structures within their branches, or a range of different branches can be found growing from one core or backbone. Even the synthesis of hyperbranched polymers can be pos. influenced by RAFT polymn., allowing end group control or control over the branching d. The creative combination of RAFT polymn. with other polymn. techniques, such as ATRP or ring-opening polymn., has extended the array of available architectures. In addn., dendrimers were incorporated either as star core or endfunctionalities. A range of synthetic chem. pathways have been utilized and combined with polymer chem., pathways such as click chem.'. These combinations have allowed the creation of novel structures. RAFT processes have been combined with natural polymers and other naturally occurring building blocks, including carbohydrates, polysaccharides, cyclodextrins, proteins and peptides. The result from the intertwining of natural and synthetic materials has resulted in the formation of hybrid biopolymers. Following these developments over the last few years, it is remarkable to see that RAFT polymn. has grown from a lab curiosity to a polymn. tool that is now been used with confidence in material design. Most of the described synthetic procedures in the literature in recent years, which incorporate RAFT polymn., have been undertaken in order to design advanced materials.
- 20Matyjaszewski, K. Atom transfer radical polymerization (ATRP): current status and future perspectives. Macromolecules 2012, 45 (10), 4015– 4039, DOI: 10.1021/ma3001719Google Scholar20https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC38XlsVaqs7w%253D&md5=350b580bd1bb46c21ba5dbfd65a6811dAtom Transfer Radical Polymerization (ATRP): Current Status and Future PerspectivesMatyjaszewski, KrzysztofMacromolecules (Washington, DC, United States) (2012), 45 (10), 4015-4039CODEN: MAMOBX; ISSN:0024-9297. (American Chemical Society)A review. Current status and future perspectives in atom transfer radical polymn. (ATRP) are presented. Special emphasis is placed on mechanistic understanding of ATRP, recent synthetic and process development, and new controlled polymer architectures enabled by ATRP. New hybrid materials based on org./inorg. systems and natural/synthetic polymers are presented. Some current and forthcoming applications are described.
- 21Synatschke, C. V.; Schallon, A.; Jérôme, V.; Freitag, R.; Müller, A. H. E. Influence of Polymer Architecture and Molecular Weight of Poly(2-(dimethylamino)ethyl methacrylate) Polycations on Transfection Efficiency and Cell Viability in Gene Delivery. Biomacromolecules 2011, 12 (12), 4247– 4255, DOI: 10.1021/bm201111dGoogle Scholar21https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC3MXhtlaqtrzL&md5=5d87903451e38527a7384072d747bb1dInfluence of Polymer Architecture and Molecular Weight of Poly(2-(dimethylamino)ethyl methacrylate) Polycations on Transfection Efficiency and Cell Viability in Gene DeliverySynatschke, Christopher V.; Schallon, Anja; Jerome, Valerie; Freitag, Ruth; Mueller, Axel H. E.Biomacromolecules (2011), 12 (12), 4247-4255CODEN: BOMAF6; ISSN:1525-7797. (American Chemical Society)Nonviral gene delivery with the help of polycations has raised considerable interest in the scientific community over the past decades. Herein, we present a systematic study on the influence of the mol. wt. and architecture of poly(2-(dimethylamino)ethyl methacrylate) (PDMAEMA) on the transfection efficiency and the cytotoxicity in CHO-K1 cells. A library of well-defined homopolymers with a linear and star-shaped topol. (3- and 5-arm stars) was synthesized via atom transfer radical polymn. (ATRP). The mol. wts. of the polycations ranged from 16 to 158 kDa. We found that the cytotoxicity at a given mol. wt. decreased with increasing no. of arms. For a successful transfection a min. mol. wt. was necessary, since the polymers with a no.-av. mol. wt., Mn, below 20 kDa showed negligible transfection efficiency at any of the tested polyelectrolyte complex compns. From the combined anal. of cytotoxicity and transfection data, we propose that polymers with a branched architecture and an intermediate mol. wt. are the most promising candidates for efficient gene delivery, since they combine low cytotoxicity with acceptable transfection results.
- 22Burgevin, F.; Hapeshi, A.; Song, J.-I.; Omedes-Pujol, M.; Christie, A.; Lindsay, C.; Perrier, S. Cationic star copolymers obtained by the arm first approach for gene transfection. Polym. Chem. 2023, 14, 3707– 3717, DOI: 10.1039/D3PY00352CGoogle ScholarThere is no corresponding record for this reference.
- 23Ros, S.; Freitag, J. S.; Smith, D. M.; Stöver, H. D. H. Charge-Shifting Polycations Based on N, N-(dimethylamino) ethyl Acrylate for Improving Cytocompatibility During DNA Delivery. ACS Omega 2020, 5 (16), 9114– 9122, DOI: 10.1021/acsomega.9b03734Google Scholar23https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BB3cXntVyit7k%253D&md5=592a1183564e7d7132577c5477df722aCharge-Shifting Polycations Based on N,N-(dimethylamino)ethyl Acrylate for Improving Cytocompatibility During DNA DeliveryRos, Samantha; Freitag, Jessica S.; Smith, David M.; Stover, Harald D. H.ACS Omega (2020), 5 (16), 9114-9122CODEN: ACSODF; ISSN:2470-1343. (American Chemical Society)Synthetic polycations are studied extensively as DNA delivery agents because of their ease of prodn., good chem. stability, and low cost relative to viral vectors. This report describes the synthesis of charge-shifting polycations based on N,N-(dimethylamino)ethyl acrylate (DMAEA) and 3-aminopropylmethacryamide (APM), called PAD copolymers, and their use for in vitro DNA delivery into HeLa cells. PAD copolymers of varying compns. were prepd. by RAFT polymn. to yield polymers of controlled mol. wts. with low dispersities. Model hydrolysis studies were carried out to assess the rate of charge-shifting of the polycations by loss of the cationic dimethylaminoethanol side chains. They showed redn. in the net cationic charge by about 10-50% depending on compn. after 2 days at pH 7, forming polyampholytes comprising permanent cationic groups, residual DMAEA, as well as anionic acrylic acid groups. HeLa cells exposed for 4 h to PAD copolymers with the greatest charge-shifting ability showed comparable or higher viability at high concns., relative to the noncharge shifting polycations PAPM and polyethyleneimine (PEI) 2 days post-exposure. Cell uptake efficiency of PAD/60bp-Cy3 DNA polyplexes at 2.5:1 N/P ratio was very high (>95%) for all compns., exceeding the uptake efficiency of PEI polyplexes of equiv. compn. These results suggest that these PAD copolymers, and in particular PAD80 contg. 80 mol % DMAEA, have suitable rates of charge-shifting hydrolysis for DNA delivery, as PAD80 showed reduced cytotoxicity at high concns., while still retaining high uptake efficiencies. In addn., the polyampholytes formed during DMAEA hydrolysis in PAD copolymers can offer enhanced long-term cytocompatibility.
- 24Cook, A. B.; Peltier, R.; Hartlieb, M.; Whitfield, R.; Moriceau, G.; Burns, J. A.; Haddleton, D. M.; Perrier, S. Cationic and hydrolysable branched polymers by RAFT for complexation and controlled release of dsRNA. Polym. Chem. 2018, 9 (29), 4025– 4035, DOI: 10.1039/C8PY00804CGoogle Scholar24https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC1cXht1Kms7rK&md5=8afc458517ca45d84065985be1275725Cationic and hydrolysable branched polymers by RAFT for complexation and controlled release of dsRNACook, Alexander B.; Peltier, Raoul; Hartlieb, Matthias; Whitfield, Richard; Moriceau, Guillaume; Burns, James A.; Haddleton, David M.; Perrier, SebastienPolymer Chemistry (2018), 9 (29), 4025-4035CODEN: PCOHC2; ISSN:1759-9962. (Royal Society of Chemistry)The controlled release of nucleic acids from cationic polymers is an important criteria for the design of gene delivery systems, and can be difficult to achieve due to the persistent pos. charges required to initially complex the nucleic acids. Here, we report the use of highly branched tertiary amine-rich polymers for the complexation and release of dsRNA over a prolonged period of time. Controlled release of dsRNA is obtained via self-catalyzed hydrolysis of the polymer side chains and assocd. change in electrostatic charge. Reversible addn.-fragmentation chain transfer (RAFT) polymn. was utilized to synthesize a series of branched polymers of 2-(dimethylamino)ethyl acrylate (DMAEA), 3-(dimethylamino)propyl acrylate (DMAPA), and 2-(dimethylamino)ethyl methacrylate (DMAEMA) (MW ∼60 000-200 000 g mol-1) and copolymers thereof. The hydrolysis kinetics of all synthesized polymer materials were followed by 1H NMR spectroscopy. Complexation with dsRNA resulted in the formation of polyplex nanoparticles (N/P ratio of 5) with sizes of approx. 400 nm and surface charges of +15 mV. An agarose gel retardation assay showed sustained release of dsRNA from p(DMAEA-co-DMAEMA) for a period of more than 2 wk. Unlike branched PEI commonly used for gene delivery, the majority of these systems showed little toxicity to cells (NIH3T3 fibroblasts). The results point towards pDMAPA and p(DMAEA-co-DMAEMA) being promising polymers for the controlled release of nucleic acids over prolonged periods.
- 25McCool, M.; Senogles, E. The self-catalysed hydrolysis of poly (N, N-dimethylaminoethyl acrylate). Eur. Polym. J. 1989, 25 (7–8), 857– 860, DOI: 10.1016/0014-3057(89)90054-2Google Scholar25https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADyaK3cXotVWj&md5=589bac9a9213f93d4e4a76809ed952a4The self-catalyzed hydrolysis of poly(N,N-dimethylaminoethyl acrylate)McCool, M. B.; Senogles, E.European Polymer Journal (1989), 25 (7-8), 857-60CODEN: EUPJAG; ISSN:0014-3057.Poly(N,N-dimethylaminoethyl acrylate) (I) is a weak base in aq. soln. and slowly hydrolyzes at ambient temp.; the reaction was monitored by 1H and 13C NMR and reacted a limiting conversion of ∼60% after 1-2 wk. Poly(N,N-diethylaminoethyl acrylate) reacted similarly and both polymers underwent ester interchange reactions in MeOH. 13C-NMR spectra of I prepd. by radical polymn. showed no evidence for any defect structures.
- 26Truong, N. P.; Jia, Z.; Burges, M.; McMillan, N. A.; Monteiro, M. J. Self-catalyzed degradation of linear cationic poly (2-dimethylaminoethyl acrylate) in water. Biomacromolecules 2011, 12 (5), 1876– 1882, DOI: 10.1021/bm200219eGoogle Scholar26https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC3MXltFGmt7s%253D&md5=abc5282a81e205e34d102cb73cd886a0Self-catalyzed degradation of linear cationic poly(2-dimethylaminoethyl acrylate) in waterTruong, Nghia P.; Jia, Zhongfan; Burges, Melinda; McMillan, Nigel A. J.; Monteiro, Michael J.Biomacromolecules (2011), 12 (5), 1876-1882CODEN: BOMAF6; ISSN:1525-7797. (American Chemical Society)We report the synthesis of a low cytotoxic polycation that maintains its cationic strength for well over a few hours then degrades into a benign polymer with nontoxic byproducts. Well-defined poly(2-dimethylaminoethyl acrylate) (PDMAEA) of five different mol. wts. prepd. using reversible addn.-fragmentation chain transfer (RAFT) "living" radical polymn. degrades slowly over 200 h (∼8 days). As this degrdn. is independent of both the polymer mol. wt. and soln. pH, it is consistent with a self-catalyzed hydrolysis process without the need for an internal or external degrdn. trigger. In addn., the polymer shows little or no cytotoxicity to HeLa cells for the mol. wts. of 5600 and below, even at very high polymer concns. (equiv. to a nitrogen/phosphorus ratio of 200). Therefore, at sufficiently low mol. wts. this polymer has the essential attributes (i.e., ability to autodegradable and low toxicity) for a delivery carrier suitable for DNA or siRNA.
- 27Truong, N. P.; Jia, Z.; Burgess, M.; Payne, L.; McMillan, N. A.; Monteiro, M. J. Self-catalyzed degradable cationic polymer for release of DNA. Biomacromolecules 2011, 12 (10), 3540– 3548, DOI: 10.1021/bm2007423Google Scholar27https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC3MXhtV2rtrbF&md5=640d3cf240d369fc9870395d051c8fd5Self-Catalyzed Degradable Cationic Polymer for Release of DNATruong, Nghia P.; Jia, Zhongfan; Burgess, Melinda; Payne, Liz; McMillan, Nigel A. J.; Monteiro, Michael J.Biomacromolecules (2011), 12 (10), 3540-3548CODEN: BOMAF6; ISSN:1525-7797. (American Chemical Society)The controlled release of siRNA or DNA complexes from cationic polymers is an important parameter design in polymer-based delivery carriers. In this work, we use the self-catalyzed degradable poly(2-dimethylaminoethyl acrylate) (PDMAEA) to strongly bind, protect, and then release oligo DNA (a mimic for siRNA) without the need for a cellular or external trigger. This self-catalyzed hydrolysis process of PDMAEA forms poly(acrylic acid) and N,N'-dimethylamino Et ethanol, both of which have little or no toxicity to cells, and offers the advantage of little or no toxicity to off-target cells and tissues. We found that PDMAEA makes an ideal component of a delivery carrier by protecting the oligo DNA for a sufficiently long period of time to transfect most cells (80% transfection after 4 h) and then has the capacity to release the DNA inside the cells after ∼10 h. The PDMAEA formed large nanoparticle complexes with oligo DNA of ∼400 nm that protected the oligo DNA from DNase in serum. The nanoparticle complexes showed no toxicity for all mol. wts. at a nitrogen/phosphorus (N/P) ratio of 10. Only the higher mol. wt. polymers at very high N/P ratios of 200 showed significant levels of cytotoxicity. These attributes make PDMAEA a promising candidate as a component in the design of a gene delivery carrier without the concern about accumulated toxicity of nanoparticles in the human body after multiadministration, an issue that has become increasingly more important.
- 28Richter, F.; Martin, L.; Leer, K.; Moek, E.; Hausig, F.; Brendel, J. C.; Traeger, A. Tuning of Endosomal Escape and Gene Expression by Functional Groups, Molecular Weight and Transfection Medium: A Structure-Activity Relationship Study. J. Mater. Chem. B 2020, 8, 5026– 5041, DOI: 10.1039/D0TB00340AGoogle ScholarThere is no corresponding record for this reference.
- 29Richter, F.; Leer, K.; Martin, L.; Mapfumo, P.; Solomun, J. I.; Kuchenbrod, M. T.; Hoeppener, S.; Brendel, J. C.; Traeger, A. The impact of anionic polymers on gene delivery: how composition and assembly help evading the toxicity-efficiency dilemma. J. Nanobiotechnol. 2021, 19, 292, DOI: 10.1186/s12951-021-00994-2Google Scholar29https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BB38XivVOnsL4%253D&md5=5ce1c855765214f4b7a0cd5fd455ce00The impact of anionic polymers on gene delivery: how composition and assembly help evading the toxicity-efficiency dilemmaRichter, Friederike; Leer, Katharina; Martin, Liam; Mapfumo, Prosper; Solomun, Jana I.; Kuchenbrod, Maren T.; Hoeppener, Stephanie; Brendel, Johannes C.; Traeger, AnjaJournal of Nanobiotechnology (2021), 19 (1), 292CODEN: JNOAAO; ISSN:1477-3155. (BioMed Central Ltd.)Cationic polymers have been widely studied for non-viral gene delivery due to their ability to bind genetic material and to interact with cellular membranes. However, their charged nature carries the risk of increased cytotoxicity and interaction with serum proteins, limiting their potential in vivo application. Therefore, hydrophilic or anionic shielding polymers are applied to counteract these effects. Herein, a series of micelle-forming and micelle-shielding polymers were synthesized via RAFT polymn. The copolymer poly[(Bu acrylate)-b-(2-(di-Me amino)ethyl acrylamide)] (P(nBA-b-DMAEAm)) was assembled into cationic micelles and different shielding polymers were applied, i.e., poly(acrylic acid) (PAA), poly(4-acryloyl morpholine) (PNAM) or P(NAM-b-AA) block copolymer. These systems were compared to a triblock terpolymer micelle comprising PAA as the middle block. The assemblies were investigated regarding their morphol., interaction with pDNA, cytotoxicity, transfection efficiency, polyplex uptake and endosomal escape. The naked cationic micelle exhibited superior transfection efficiency, but increased cytotoxicity. The addn. of shielding polymers led to reduced toxicity. In particular, the triblock terpolymer micelle convinced with high cell viability and no significant loss in efficiency. The highest shielding effect was achieved by layering micelles with P(NAM-b-AA) supporting the colloidal stability at neutral zeta potential and completely restoring cell viability while maintaining moderate transfection efficiencies. The high potential of this micelle-layer-combination for gene delivery was illustrated for the first time.
- 30Truong, N. P.; Gu, W.; Prasadam, I.; Jia, Z.; Crawford, R.; Xiao, Y.; Monteiro, M. J. An influenza virus-inspired polymer system for the timed release of siRNA. Nat. Commun. 2013, 4 (1), 1902, DOI: 10.1038/ncomms2905Google Scholar30https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A280%3ADC%252BC3snlvFCqtw%253D%253D&md5=a9c0ca3205517013c3f753b32a43e482An influenza virus-inspired polymer system for the timed release of siRNATruong Nghia P; Gu Wenyi; Prasadam Indira; Jia Zhongfan; Crawford Ross; Xiao Yin; Monteiro Michael JNature communications (2013), 4 (), 1902 ISSN:.Small interfering RNA silences specific genes by interfering with mRNA translation, and acts to modulate or inhibit specific biological pathways; a therapy that holds great promise in the cure of many diseases. However, the naked small interfering RNA is susceptible to degradation by plasma and tissue nucleases and due to its negative charge unable to cross the cell membrane. Here we report a new polymer carrier designed to mimic the influenza virus escape mechanism from the endosome, followed by a timed release of the small interfering RNA in the cytosol through a self-catalyzed polymer degradation process. Our polymer changes to a negatively charged and non-toxic polymer after the release of small interfering RNA, presenting potential for multiple repeat doses and long-term treatment of diseases.
- 31Tran, N. T.; Truong, N. P.; Gu, W.; Jia, Z.; Cooper, M. A.; Monteiro, M. J. Timed-release polymer nanoparticles. Biomacromolecules 2013, 14 (2), 495– 502, DOI: 10.1021/bm301721kGoogle Scholar31https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC3sXkt1yguw%253D%253D&md5=f04eb155e4ff9c77681803ac317585bfTimed-Release Polymer NanoparticlesTran, Nguyen T. D.; Truong, Nghia P.; Gu, Wenyi; Jia, Zhongfan; Cooper, Matthew A.; Monteiro, Michael J.Biomacromolecules (2013), 14 (2), 495-502CODEN: BOMAF6; ISSN:1525-7797. (American Chemical Society)Triggered-release of encapsulated therapeutics from nanoparticles without remote or environmental triggers was demonstrated in this work. Disassembly of the polymer nanoparticles to unimers at precise times allowed the controlled release of oligo DNA. The polymers used in this study consisted of a hydrophilic block for stabilization and second thermoresponsive block for self-assembly and disassembly. At temps. below the second block's LCST (i.e., below 37 °C for in vitro assays), the diblock copolymer was fully water-sol., and when heated to 37 °C, the polymer self-assembled into a narrow size distribution of nanoparticles with an av. diam. of approx. 25 nm. The thermoresponsive nature of the second block could be manipulated in situ by the self-catalyzed degrdn. of cationic 2-(dimethylamino)ethyl acrylate (DMAEA) units to neg. charged acrylic acid groups and when the amt. of acid groups was sufficiently high to increase the LCST of the second block above 37 °C. The disassembly of the nanoparticles could be controlled from 10 to 70 h. The use of these nanoparticles as a combined therapy, in which one or more agents can be released in a predetd. way, has the potential to improve the personal point of care treatment of patients.
- 32Tran, N. T.; Jia, Z.; Truong, N. P.; Cooper, M. A.; Monteiro, M. J. Fine tuning the disassembly time of thermoresponsive polymer nanoparticles. Biomacromolecules 2013, 14 (10), 3463– 3471, DOI: 10.1021/bm4007858Google Scholar32https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC3sXhsVCgtbbP&md5=e1252dea2d4bd6ec81784d2a3a67cd78Fine Tuning the Disassembly Time of Thermoresponsive Polymer Nanoparticles.Tran, Nguyen T. D.; Jia, Zhongfan; Truong, Nghia P.; Cooper, Matthew A.; Monteiro, Michael J.Biomacromolecules (2013), 14 (10), 3463-3471CODEN: BOMAF6; ISSN:1525-7797. (American Chemical Society)Timed-released disassembly of nanoparticles without a remote trigger or environmental cues is demonstrated in this work. The reversible addn.-fragmentation chain transfer (RAFT) polymn. allowed the fine-tuning of the chem. compn. in the diblock copolymers, in which the first block consisted of a hydrophilic monomer (DMA) and the second random block consisted of three different monomers: (a) the thermoresponsive NIPAM, (b) the self-catalyzed hydrolyzable DMAEA, and (c) the hydrophobic BA. These diblock copolymers were solubilized in water below the lower crit. soln. temp. (LCST) of the thermoresponsive second block, and heated to 37 °C (i.e., >LCST) to form small micelle nanoparticles with a narrow particle size distribution. As DMAEA hydrolyzed to acrylic acid groups, the LCST of the diblock increased, and the time at the start of micelle disassembly (tstart) corresponded to the point where the LCST was equal to the soln. temp. (i.e., 37 °C). The high water content in the PNIPAM core allowed an even degrdn. of the core over time. The copolymer compn. allowed fine control over tstart, as this time was linearly dependent upon the BA units in the second block. These nanoparticles could also be designed to be stable (i.e., not disassemble) over a wide pH range or disassemble below a pH of 7.3. Addnl., the time from the start of disassembly to full unimer formation (tdegrade) could be controlled by the amt. of DMAEA units in the second block. A longer tdegrade (∼5.5 h) was found when the no. of DMAEA units was 42 compared to tdegrade of 1.1 h for 25 units. The nanoparticles designed in this work, through fine control of the polymer chem. compn., have the potential for drug delivery purposes for timed-release of drugs and prodrugs and other wide-ranging applications where timed-release would be beneficial.
- 33Solomun, J. I.; Cinar, G.; Mapfumo, P.; Richter, F.; Moek, E.; Hausig, F.; Martin, L.; Hoeppener, S.; Nischang, I.; Traeger, A. Solely aqueous formulation of hydrophobic cationic polymers for efficient gene delivery. Int. J. Pharm. 2021, 593, 120080, DOI: 10.1016/j.ijpharm.2020.120080Google Scholar33https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BB3cXisFSnsb7I&md5=bc64bd44d33d1a5bf1cc69d27aa4a7a1Solely aqueous formulation of hydrophobic cationic polymers for efficient gene deliverySolomun, Jana I.; Cinar, Gizem; Mapfumo, Prosper; Richter, Friederike; Moek, Elisabeth; Hausig, Franziska; Martin, Liam; Hoeppener, Stephanie; Nischang, Ivo; Traeger, AnjaInternational Journal of Pharmaceutics (Amsterdam, Netherlands) (2021), 593 (), 120080CODEN: IJPHDE; ISSN:0378-5173. (Elsevier B.V.)Cationic polymers are promising gene delivery vectors due to their ability to bind and protect genetic material. The introduction of hydrophobic moieties into cationic polymers can further improve the vector efficiency, but common formulations of hydrophobic polymers involve harsh conditions such as org. solvents, impairing intactness and loading efficiency of the genetic material. In this study, a mild, aq. formulation method for the encapsulation of high amts. of genetic material is presented. A well-defined pH-responsive hydrophobic copolymer, i.e. poly((n-butylmethacrylate)-co-(methylmethacrylate)-co-(2-(dimethylamino) ethylmethacrylate)), (PBMD) was synthesized by reversible addn. fragmentation chain transfer (RAFT) polymn. Exploiting the pH-dependent soly. behavior of the polymer, stable pDNA loaded nanoparticles were prepd. and characterized using anal. ultracentrifugation (AUC), cryo-transmission electron microscopy (cryo-TEM) and dynamic light scattering (DLS). This novel formulation approach showed high transfection efficiencies in HEK293T cells, while requiring 5- to 10-fold less pDNA compared to linear polyethylenimine (LPEI), in particular at short incubation times and in serum-contg. media. Furthermore, the formulation was successfully adopted for siRNA and mRNA encapsulation and the com. approved polymer Eudragit E(PO/100). Overall, the aq. formulation approach, accompanied by a tailor-made hydrophobic polymer and detailed physicochem. and application studies, led to improved gene delivery vectors with high potential for further applications.
- 34Adolph, E. J.; Nelson, C. E.; Werfel, T. A.; Guo, R.; Davidson, J. M.; Guelcher, S. A.; Duvall, C. L. Enhanced Performance of Plasmid DNA Polyplexes Stabilized by a Combination of Core Hydrophobicity and Surface PEGylation. J. Mater. Chem. B 2014, 2 (46), 8154– 8164, DOI: 10.1039/C4TB00352GGoogle ScholarThere is no corresponding record for this reference.
- 35Convertine, A. J.; Benoit, D. S.; Duvall, C. L.; Hoffman, A. S.; Stayton, P. S. Development of a novel endosomolytic diblock copolymer for siRNA delivery. J. Controlled Release 2009, 133 (3), 221– 229, DOI: 10.1016/j.jconrel.2008.10.004Google Scholar35https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BD1MXpsF2ktQ%253D%253D&md5=e46b2da9dca2388591ffd43dde0bd26bDevelopment of a novel endosomolytic diblock copolymer for siRNA deliveryConvertine, Anthony J.; Benoit, Danielle S. W.; Duvall, Craig L.; Hoffman, Allan S.; Stayton, Patrick S.Journal of Controlled Release (2009), 133 (3), 221-229CODEN: JCREEC; ISSN:0168-3659. (Elsevier B.V.)The gene knockdown activity of small interfering RNA (siRNA) has led to their use as target validation tools and as potential therapeutics for a variety of diseases. The delivery of these double-stranded RNA macromols. has proven to be challenging, however, and in many cases, is a barrier to their deployment. Here we report the development of a new diblock copolymer family that was designed to enhance the systemic and intracellular delivery of siRNA. These diblock copolymers were synthesized using the controlled reversible addn. fragmentation chain transfer polymn. (RAFT) method and are composed of a pos.-charged block of dimethylaminoethyl methacrylate (DMAEMA) to mediate siRNA condensation, and a second endosomal-releasing block composed of DMAEMA and propylacrylic acid (PAA) in roughly equimolar ratios, together with Bu methacrylate (BMA). A related series of diblock compns. were characterized, with the cationic block kept const., and with the ratio of DMAEMA and PAA to BMA varied. These carriers became sharply hemolytic at endosomal pH regimes, with increasing hemolytic activity seen as the percentage of BMA in the second block was systematically increased. The diblock copolymers condensed siRNA into 80-250 nm particles with slightly pos. Zeta potentials. SiRNA-mediated knockdown of a model protein, namely glyceraldehyde 3-phosphate dehydrogenase (GAPDH), in HeLa cells generally followed the hemolytic activity trends, with the most hydrophobic second block (highest BMA content) exhibiting the best knockdown. This pH-responsive carrier designed to mediate endosomal release shows significant promise for the intracellular delivery of siRNA.
- 36Liu, Z.; Zhang, Z.; Zhou, C.; Jiao, Y. Hydrophobic modifications of cationic polymers for gene delivery. Prog. Polym. Sci. 2010, 35 (9), 1144– 1162, DOI: 10.1016/j.progpolymsci.2010.04.007Google Scholar36https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC3cXhtV2isbjI&md5=6ee9ad8ed0fa21a2cce280effb33c3a2Hydrophobic modifications of cationic polymers for gene deliveryLiu, Zonghua; Zhang, Ziyong; Zhou, Changren; Jiao, YanpengProgress in Polymer Science (2010), 35 (9), 1144-1162CODEN: PRPSB8; ISSN:0079-6700. (Elsevier Ltd.)A review. Cationic polymers are the subject of intense research as non-viral gene delivery systems, due to their flexible properties, facile synthesis, robustness, and proven gene delivery efficiency. Nevertheless, low transfection efficiency and undesirable cytotoxicity remain the most challenging aspects of these cationic polymers. To overcome the disadvantages, various modifications were made to improve their gene delivery efficacy. Among them, hydrophobic modifications of the cationic polymers are receiving more and more attention. Most studies have shown that incorporation of hydrophobic chains can improve gene delivery efficiency, mainly explained by hydrophobic interaction conferred to the resulting amphiphilic polycation derivs. and by the enhanced cellular uptake by the hydrophobic chains via the lipophilic cell membrane. This review discusses recent studies on the hydrophobic modifications of cationic polymers for gene delivery. The effects of the hydrophobic modifications are discussed in terms of crit. issues in the gene delivery process, such as gene encapsulation, adsorption to cell membrane, serum inhibition, gene dissocn., cytotoxicity, and tissue-targeting. Moreover, various hydrophobic modifications of the main cationic polymeric gene carriers (polyethylenimine, chitosan, polylysine, etc.) are described with regards to the resulting gene delivery activity. The structure-function relationships discussed here provide important information and insight for the design of novel gene vectors.
- 37Eliyahu, H.; Makovitzki, A.; Azzam, T.; Zlotkin, A.; Joseph, A.; Gazit, D.; Barenholz, Y.; Domb, A. Novel dextran-spermine conjugates as transfecting agents: comparing water-soluble and micellar polymers. Gene Ther. 2005, 12 (6), 494– 503, DOI: 10.1038/sj.gt.3302395Google Scholar37https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BD2MXitV2ju7g%253D&md5=fb5a1159e66560b7ecce2f6a5418938cNovel dextran-spermine conjugates as transfecting agents: comparing water-soluble and micellar polymersEliyahu, H.; Makovitzki, A.; Azzam, T.; Zlotkin, A.; Joseph, A.; Gazit, D.; Barenholz, Y.; Domb, A. J.Gene Therapy (2005), 12 (6), 494-503CODEN: GETHEC; ISSN:0969-7128. (Nature Publishing Group)Recently, a novel cationic polymer, dextran-spermine (D-SPM) was developed for gene delivery. An efficient transfection was obtained using this polycation for a variety of genes and cell lines in serum-free or serum-poor medium. However, transfection using the water-sol. D-SPM-based polyplexes decreased with increasing serum concn. in cell culture in a concn.-dependent manner, reaching 95% inhibition at 50% serum in the cell growth medium. In order to overcome this obstacle, oleyl derivs. of D-SPM (which form micelles in aq. phase) were synthesized at 1, 10, and 20 mol% of oleyl moiety to polymer .vepsiln.-NH2 to form N-oleyl-D-SPM (ODS). Polyplexes based on ODS transfected well in medium contg. 50% serum. Comparison with polyplexes based on well-established polymers (branched and linear polyethyleneimine) and with DOTAP/Cholesterol lipoplexes showed that regarding β-galactosidase transgene expression level and cytotoxicity in tissue culture, the D-SPM and ODS compare well with the above polyplexes and lipoplexes. Intracellular trafficking using FITC-labeled ODS and Rhodamine-labeled pGeneGrip plasmid cloned with hBMP2 monitored by confocal microscopy revealed that during the transfection process the fluorescent-labeled polymer concs. in the Golgi app. and around the nucleus, while the cell cytoplasm was free of fluorescent particles, suggesting that the polyplexes move in the cell toward the nucleus by vesicular transport through the cytoplasm and not by a random diffusion. We found that the plasmids penetrate the cell nucleus without the polymer. Preliminary results in zebra fish and mice demonstrate the potential of ODS to serve as an efficient nonviral vector for in vivo transfection.
- 38Mapfumo, P. P.; Reichel, L. S.; Hoeppener, S.; Traeger, A. Improving Gene Delivery: Synergy Between Alkyl Chain Length And Lipoic Acid for Pdmaema Hydrophobic Copolymers. Macromol. Rapid Commun. 2024, 45, 2300649, DOI: 10.1002/marc.202300649Google ScholarThere is no corresponding record for this reference.
- 39Mapfumo, P. P.; Solomun, J. I.; Becker, F.; Moek, E.; Leiske, M. N.; Rudolph, L. K.; Brendel, J. C.; Traeger, A. Vitamin B3 Containing Polymers for Nanodelivery. Macromol. Biosci. 2024, 2400002, DOI: 10.1002/mabi.202400002Google ScholarThere is no corresponding record for this reference.
- 40Standard, I. 10993-5 Biological Evaluation of Medical Devices. Tests for in Vitro Cytotoxicity; Geneva, Switzerland: International Organization for Standardization, 2009; p 194.Google ScholarThere is no corresponding record for this reference.
- 41Leer, K.; Reichel, L. S.; Kimmig, J.; Richter, F.; Hoeppener, S.; Brendel, J. C.; Zechel, S.; Schubert, U. S.; Traeger, A. Optimization of Mixed Micelles Based on Oppositely Charged Block Copolymers by Machine Learning for Application in Gene Delivery. Small 2024, 20, 2306116, DOI: 10.1002/smll.202306116Google ScholarThere is no corresponding record for this reference.
- 42Patel, P.; Ibrahim, N. M.; Cheng, K. The importance of apparent pKa in the development of nanoparticles encapsulating siRNA and mRNA. Trends Pharmacol. Sci. 2021, 42 (6), 448– 460, DOI: 10.1016/j.tips.2021.03.002Google Scholar42https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BB3MXns1aqtL0%253D&md5=151ad19fba69fb7f0be6cd37fbbc9376The Importance of Apparent pKa in the Development of Nanoparticles Encapsulating siRNA and mRNAPatel, Pratikkumar; Ibrahim, Nurudeen Mohammed; Cheng, KunTrends in Pharmacological Sciences (2021), 42 (6), 448-460CODEN: TPHSDY; ISSN:0165-6147. (Elsevier Ltd.)A review. Polymer and lipid nanoparticles have been extensively used as carriers to address the biol. barriers encountered in siRNA and mRNA delivery. We summarize the crucial role of nanoparticle charge and ionizability in complexing RNAs, binding to biol. components, escaping from the endosome, and releasing RNAs into the cytoplasm. We highlight the significant impact of the apparent pKa of nanoparticles on their efficacy and toxicity, and the importance of optimizing pKa in the development of lead formulations for RNAs. We also discuss the feasibility of fine-tuning the pKa in nanoparticles and the applications of this approach in the optimization of delivery systems for RNAs.
- 43Alabi, C. A.; Love, K. T.; Sahay, G.; Yin, H.; Luly, K. M.; Langer, R.; Anderson, D. G. Multiparametric approach for the evaluation of lipid nanoparticles for siRNA delivery. Proc. Natl. Acad. Sci. U.S.A. 2013, 110 (32), 12881– 12886, DOI: 10.1073/pnas.1306529110Google Scholar43https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC3sXhsVShurbN&md5=f2bc33e036370b72cab58dd701a364b6Multiparametric approach for the evaluation of lipid nanoparticles for siRNA deliveryAlabi, Christopher A.; Love, Kevin T.; Sahay, Gaurav; Yin, Hao; Luly, Kathryn M.; Langer, Robert; Anderson, Daniel G.Proceedings of the National Academy of Sciences of the United States of America (2013), 110 (32), 12881-12886, S12881/1-S12881/22CODEN: PNASA6; ISSN:0027-8424. (National Academy of Sciences)Nanoparticle-mediated siRNA delivery is a complex process that requires transport across numerous extracellular and intracellular barriers. As such, the development of nanoparticles for efficient delivery would benefit from an understanding of how parameters assocd. with these barriers relate to the physicochem. properties of nanoparticles. Here, we use a multiparametric approach for the evaluation of lipid nanoparticles (LNPs) to identify relationships between structure, biol. function, and biol. activity. Our results indicate that evaluation of multiple parameters assocd. with barriers to delivery such as siRNA entrapment, pKa, LNP stability, and cell uptake as a collective may serve as a useful prescreening tool for the advancement of LNPs in vivo. This multiparametric approach complements the use of in vitro efficacy results alone for prescreening and improves in vitro and in vivo translation by minimizing false negatives. For the LNPs used in this work, the evaluation of multiple parameters enabled the identification of LNP pKa as one of the key determinants of LNP function and activity both in vitro and in vivo. It is anticipated that this type of anal. can aid in the identification of meaningful structure-function-activity relationships, improve the in vitro screening process of nanoparticles before in vivo use, and facilitate the future design of potent nanocarriers.
- 44Appleby, C. A.; Wittenberg, B. A.; Wittenberg, J. B. Nicotinic acid as a ligand affecting leghemoglobin structure and oxygen reactivity. Proc. Natl. Acad. Sci. U.S.A. 1973, 70 (2), 564– 568, DOI: 10.1073/pnas.70.2.564Google ScholarThere is no corresponding record for this reference.
- 45Ros, S.; Wang, J.; Burke, N. A.; Stöver, H. D. H. A Mechanistic Study of the Hydrolysis of Poly [N, N-(dimethylamino) ethyl acrylates] as Charge-Shifting Polycations. Macromolecules 2020, 53 (9), 3514– 3523, DOI: 10.1021/acs.macromol.9b02272Google Scholar45https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BB3cXntlert7Y%253D&md5=22b718d55109bebc24cc7fcd4426b234A Mechanistic Study of the Hydrolysis of Poly[N,N-(dimethylamino)ethyl acrylates] as Charge-Shifting PolycationsRos, Samantha; Wang, Jiexi; Burke, Nicholas A. D.; Stover, Harald D. H.Macromolecules (Washington, DC, United States) (2020), 53 (9), 3514-3523CODEN: MAMOBX; ISSN:0024-9297. (American Chemical Society)Polycations are used extensively in applications ranging from enhanced oil recovery to biomaterials. Poly[N,N-(dimethylamino)ethyl acrylate] (PDMAEA) has attracted interest for biomaterial applications because of its rapid hydrolysis; however, the mechanism of hydrolysis and the conditions that affect degrdn. often appear amiss in the field. In this report, a detailed 1H NMR spectroscopy study of the hydrolysis of PDMAEA was carried out between pH 0 and 14. In contrast to a widely held view, the rates of hydrolysis of this polymer were found to be highly pH-dependent with half-lives varying from years to minutes as a function of pH. The extent of hydrolysis was also found to be pH-dependent, with a distinct plateau at about 50-60% hydrolysis at pH 7 due to the electrostatic repulsion of anionic carboxylate groups and hydroxide. This was contrasted with the acid-catalyzed mechanism where hydrolysis of PDMAEA at pH 0.3 did not show a plateau in the extent of hydrolysis, reaching 88% hydrolysis after 8 days at 70°C. In addn., the effects of neighboring functional groups on DMAEA hydrolysis in copolymers were explored, with anionic, neutral/hydrophilic, and cationic comonomers found to affect the rates of hydrolysis up to 20-fold at pH 7. Finally, two novel analogs of DMAEA with an addnl. dimethylamino group in different positions of the side chain were synthesized and polymd. to probe the effect of this added tertiary amine on the hydrolysis of the ester linkages. Poly[1,3-bis (dimethylamino)-2-Pr acrylate] (PBDMAPA) hydrolyzed more than 500 times faster at pH 7 than its linear isomer poly[2-((2-(dimethylamino)ethyl)(methyl)amino)ethyl acrylate] (PDEMEA). These results further highlight the importance of the intramol. interactions of the dimethylamino substituent of PDMAEA and the effect of proximity and steric hindrance of the amino group as well as neighboring functional groups of comonomers.
- 46LePecq, J.-B.; Paoletti, C. A fluorescent complex between ethidium bromide and nucleic acids: physical─chemical characterization. J. Mol. Biol. 1967, 27 (1), 87– 106, DOI: 10.1016/0022-2836(67)90353-1Google ScholarThere is no corresponding record for this reference.
- 47Olmsted, J., III; Kearns, D. R. Mechanism of ethidium bromide fluorescence enhancement on binding to nucleic acids. Biochemistry 1977, 16 (16), 3647– 3654, DOI: 10.1021/bi00635a022Google Scholar47https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADyaE2sXltFegtLw%253D&md5=7dd2868d98cff0cfb0be2735ca90454bMechanism of ethidium bromide fluorescence enhancement on binding to nucleic acidsOlmsted, John, III; Kearns, David R.Biochemistry (1977), 16 (16), 3647-54CODEN: BICHAW; ISSN:0006-2960.From an examn. of the effect of different solvents on the fluorescence lifetime, quenching of fluorescence by proton acceptors, and the substantial lengthening of lifetime obsd. upon deuteration of the amino protons, regardless of the medium, it appears that proton transfer from the excited singlet state is the process primarily responsible for the low fluorescence yield in most polar solvents. Enhancement of fluorescence upon intercalation is attributed to a redn. in the rate of excited state proton transfer to solvent mols. The proposed mechanism accounts for the ∼3.5-fold increase in the lifetime of free ethidium bromide in going from H2O to D2O; the fact that addn. of small amts. of H2O to nonaq. solvents decreases the fluorescence, whereas addn. of small amts. of D2O enhances the fluorescence; and the enhancement of the ethidium bromide triplet state yield on binding to DNA.
- 48Richter, F.; Mapfumo, P.; Martin, L.; Solomun, J. I.; Hausig, F.; Frietsch, J. J.; Ernst, T.; Hoeppener, S.; Brendel, J. C.; Traeger, A. Improved gene delivery to K-562 leukemia cells by lipoic acid modified block copolymer micelles. J. Nanobiotechnol. 2021, 19 (1), 70, DOI: 10.1186/s12951-021-00801-yGoogle ScholarThere is no corresponding record for this reference.
- 49Prevette, L. E.; Kodger, T. E.; Reineke, T. M.; Lynch, M. L. Deciphering the role of hydrogen bonding in enhancing pDNA- polycation interactions. Langmuir 2007, 23 (19), 9773– 9784, DOI: 10.1021/la7009995Google Scholar49https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BD2sXpt1Wisrk%253D&md5=47380c5628d5e8e7525f1d5c036778a7Deciphering the Role of Hydrogen Bonding in Enhancing pDNA-Polycation InteractionsPrevette, Lisa E.; Kodger, Tom E.; Reineke, Theresa M.; Lynch, Matthew L.Langmuir (2007), 23 (19), 9773-9784CODEN: LANGD5; ISSN:0743-7463. (American Chemical Society)There is considerable interest in the binding and condensation of DNA with polycations to form polyplexes because of their possible application to cellular nucleic acid delivery. This work focuses on studying the binding of plasmid DNA (pDNA) with a series of poly(glycoamidoamine)s (PGAAs) that have previously been shown to deliver pDNA in vitro in an efficient and nontoxic manner. Herein, we examine the PGAA-pDNA binding energetics, binding-linked protonation, and electrostatic contribution to the free energy with isothermal titrn. calorimetry (ITC). The size and charge of the polyplexes at various ITC injection points were then investigated by light scattering and ζ-potential measurements to provide comprehensive insight into the formation of these polyplexes. An anal. of the calorimetric data revealed a three-step process consisting of two different endothermic contributions followed by the condensation/aggregation of polyplexes. The strength of binding and the point of charge neutralization were found to be dependent upon the hydroxyl stereochem. of the carbohydrate moiety within each polymer repeat unit. CD spectra reveal that the PGAAs induce pDNA secondary structure changes upon binding, which suggest a direct interaction between the polymers and the DNA base pairs. IR spectroscopy expts. confirmed both base pair and phosphate group interactions and, more specifically, showed that the stronger-binding PGAAs had more pronounced interactions at both sites. Thus, we conclude that the mechanism of poly(glycoamidoamine)-pDNA binding is most likely a combination of electrostatics and hydrogen bonding in which long-range Coulombic forces initiate the attraction and hydroxyl groups in the carbohydrate comonomer, depending on their stereochem., further enhance the assocn. through hydrogen bonding to the DNA base pairs.
- 50Kong, D. C.; Yang, M. H.; Zhang, X. S.; Du, Z. C.; Fu, Q.; Gao, X. Q.; Gong, J. W. Control of polymer properties by entanglement: a review. Macromol. Mater. Eng. 2021, 306 (12), 2100536, DOI: 10.1002/mame.202100536Google Scholar50https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BB3MXitFans7bJ&md5=d090e9f4b0fe0f536244b08ace880701Control of Polymer Properties by Entanglement: A ReviewKong, De-Chao; Yang, Ming-Hao; Zhang, Xue-Song; Du, Zu-Chen; Fu, Qiang; Gao, Xue-Qin; Gong, Jia-WeiMacromolecular Materials and Engineering (2021), 306 (12), 2100536CODEN: MMENFA; ISSN:1438-7492. (Wiley-VCH Verlag GmbH & Co. KGaA)A review. Chain entanglement, either cohesional or topol., distinguishes polymers from other engineering materials. It impedes the movement of mol. segments and influences the polymer rheol., morphol., and mech. properties. Although a high level of entanglement can increase the polymer toughness, excessive entanglement should be avoided because it causes a high melt viscosity making the processing difficult. This review tended to elucidate the influence of entanglement on the polymer structure, detg. the material properties and processability. A wide range of methods used to fine control the degrees of chain entanglement are summarized. The methods are applicable to polymers in solns., melts, and condensed states with advantages and limitations discussed in detail. The authors also examd. the effect of the entanglement on polymer crystn.-the mechanism remains a controversial issue. This review will provide general guidance to designing and processing polymer materials with desired properties via a rational route of controlling the chain entanglement.
- 51Koltzenburg, S.; Maskos, M.; Nuyken, O.; Mülhaupt, R. Polymere: Synthese, Eigenschaften und Anwendungen; Springer, 2014.Google ScholarThere is no corresponding record for this reference.
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- 53Bus, T.; Traeger, A.; Schubert, U. S. The great escape: how cationic polyplexes overcome the endosomal barrier. J. Mater. Chem. B 2018, 6 (43), 6904– 6918, DOI: 10.1039/C8TB00967HGoogle Scholar53https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC1cXhslSrsLfJ&md5=62d63fe76a4b75545afeaa170bffce43The great escape: how cationic polyplexes overcome the endosomal barrierBus, Tanja; Traeger, Anja; Schubert, Ulrich S.Journal of Materials Chemistry B: Materials for Biology and Medicine (2018), 6 (43), 6904-6918CODEN: JMCBDV; ISSN:2050-7518. (Royal Society of Chemistry)A review. The targeted and efficiency-oriented delivery of (therapeutic) nucleic acids raises hope for successful gene therapy, i.e., for the local and individual treatment of acquired and inherited genetic disorders. Despite promising achievements in the field of polymer-mediated gene delivery, the efficiency of the non-viral vectors remains orders of magnitude lower than viral-mediated ones. Several obstacles on the mol. and cellular level along the gene delivery process were identified, starting from the design and formulation of the nano-sized carriers up to the targeted release to their site of action. In particular, the efficient escape from endo-lysosomal compartments was demonstrated to be a major barrier and its exact mechanism still remains unclear. Different hypotheses and theories of the endosomal escape were postulated. The most popular one is the so-called "proton sponge" hypothesis, claiming an escape by rupture of the endosome through osmotic swelling. It was the first effort to explain the excellent transfection efficiency of poly(ethylene imine). Moreover, it was thought that a unique mechanism based on the ability to capture protons and to buffer the endosomal pH is the basis of endosomal escape. Recent theories deal with the direct interaction of the cationic polyplex or free polymer with the exoplasmic lipid leaflet causing membrane destabilization, permeability or polymer-supported nanoscale hole formation. Both escape strategies are more related to viral-mediated escape compared to the "proton sponge" effect. This review addresses the different endosomal release theories and highlights their key mechanism.
- 54Chen, J.; Wang, K.; Wu, J.; Tian, H.; Chen, X. Polycations for gene delivery: dilemmas and solutions. Bioconj. Chem. 2019, 30 (2), 338– 349, DOI: 10.1021/acs.bioconjchem.8b00688Google ScholarThere is no corresponding record for this reference.
- 55Chen, J.; Tian, H.; Dong, X.; Guo, Z.; Jiao, Z.; Li, F.; Kano, A.; Maruyama, A.; Chen, X. Effective Tumor Treatment by VEGF si RNA Complexed with Hydrophobic Poly (A mino Acid)-M odified Polyethylenimine. Macromol. Biosci. 2013, 13 (10), 1438– 1446, DOI: 10.1002/mabi.201300211Google ScholarThere is no corresponding record for this reference.
- 56Auwerx, J. The human leukemia cell line, THP-1: a multifacetted model for the study of monocyte-macrophage differentiation. Experientia 1991, 47 (1), 22– 31, DOI: 10.1007/BF02041244Google Scholar56https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A280%3ADyaK3M7ltlCntA%253D%253D&md5=ab4f8ff9f199122d1abcfa85c7b4ee53The human leukemia cell line, THP-1: a multifacetted model for the study of monocyte-macrophage differentiationAuwerx JExperientia (1991), 47 (1), 22-31 ISSN:0014-4754.THP-1 is a human monocytic leukemia cell line. After treatment with phorbol esters, THP-1 cells differentiate into macrophage-like cells which mimic native monocyte-derived macrophages in several respects. Compared to other human myeloid cell lines, such as HL-60, U937, KG-1, or HEL cell lines, differentiated THP-1 cells behave more like native monocyte-derived macrophages. Because of these characteristics, the THP-1 cell line provides a valuable model for studying the mechanisms involved in macrophage differentiation, and for exploring the regulation of macrophage-specific genes as they relate to physiological functions displayed by these cells.
- 57Im, D. J.; Jeong, S. N. Transfection of Jurkat T cells by droplet electroporation. Biochem. Eng. J. 2017, 122, 133– 140, DOI: 10.1016/j.bej.2017.03.010Google Scholar57https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC2sXkvVyrur0%253D&md5=d6e235703f2120a35bf06a9ef4687a1dTransfection of Jurkat T cells by droplet electroporationIm, Do Jin; Jeong, Su-NamBiochemical Engineering Journal (2017), 122 (), 133-140CODEN: BEJOFV; ISSN:1369-703X. (Elsevier B.V.)A droplet electroporation system has been successfully demonstrated for transfection of Jurkat T cell with much higher transfection efficiency (66%) than that of conventional system (11.4%) with the advantages of superior cell viability, comparable throughput, and user-friendly interface. Because the productivity of the proposed system is much greater than previous microfluidic systems, flow cytometry is used for the anal. of transfection efficiency. The high transfection efficiency and cell viability of the droplet electroporation system is mainly attributed to small size which requires lower voltage and provides concd. environment. The successful transfection of Jurkat cell using the droplet electroporation system broadens the applicability of the proposed technol. to medical field. The implication of the present work and the future development direction for a fully automated cell engineering platform are discussed.
- 58Abraham, R. T.; Weiss, A. Jurkat T cells and development of the T-cell receptor signalling paradigm. Nat. Rev. Immunol. 2004, 4 (4), 301– 308, DOI: 10.1038/nri1330Google Scholar58https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BD2cXis1GksLY%253D&md5=adc35efbd4d1c9f181ef3c5e262102a7Jurkat T cells and development of the T-cell receptor signalling paradigmAbraham, Robert T.; Weiss, ArthurNature Reviews Immunology (2004), 4 (4), 301-308CODEN: NRIABX; ISSN:1474-1733. (Nature Publishing Group)A review. Twenty years of investigation have yielded a detailed view of the signalling machinery engaged by T-cell receptors (TCRs). Many of the fundamental insights into TCR signalling came from studies carried out with transformed T-cell lines. Perhaps the best known of these model systems is the Jurkat leukemic T-cell line, and here we review some of the key advances in the field of TCR signalling that were made with Jurkat T cells as the host.
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- 63Salem, H. A.; Wadie, W. Effect of niacin on inflammation and angiogenesis in a murine model of ulcerative colitis. Sci. Rep. 2017, 7 (1), 7139, DOI: 10.1038/s41598-017-07280-yGoogle Scholar63https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A280%3ADC%252BC1cfjs1OmtQ%253D%253D&md5=2985155e9434d34a29446bd3982a8d62Effect of Niacin on Inflammation and Angiogenesis in a Murine Model of Ulcerative ColitisSalem Hesham Aly; Wadie WalaaScientific reports (2017), 7 (1), 7139 ISSN:.Butyrate and niacin are produced by gut microbiota, however butyrate has received most attention for its effects on colonic health. The present study aimed at exploring the effect of niacin on experimental colitis as well as throwing some light on the ability of niacin to modulate angiogenesis which plays a crucial role of in the pathogenesis of inflammatory bowel disease. Rats were given niacin for 2 weeks. On day 8, colitis was induced by intrarectal administration of iodoacetamide. Rats were sacrificed on day 15 and colonic damage was assessed macroscopically and histologically. Colonic myeloperoxidase (MPO), tumour necrosis factor (TNF)-α, interleukin (IL)-10, vascular endothelial growth factor (VEGF), angiostatin and endostatin levels were determined. Niacin attenuated the severity of colitis as demonstrated by a decrease in weight loss, colonic wet weight and MPO activity. Iodoacetamide-induced rise in the colonic levels of TNF-α, VEGF, angiostatin and endostatin was reversed by niacin. Moreover, niacin normalized IL-10 level in colon. Mepenzolate bromide, a GPR109A receptor blocker, abolished the beneficial effects of niacin on body weight, colon wet weight as well as colonic levels of MPO and VEGF. Therefore, niacin was effective against iodoacetamide-induced colitis through ameliorating pathologic angiogenesis and inflammatory changes in a GPR109A-dependent manner.
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- 66Aggarwal, B. B.; Gupta, S. C.; Sung, B. Curcumin: an orally bioavailable blocker of TNF and other pro-inflammatory biomarkers. Br. J. Pharmacol. 2013, 169 (8), 1672– 1692, DOI: 10.1111/bph.12131Google Scholar66https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC3sXht1elsrnL&md5=3f052e73279fd5eebb2341057dbb7f6bCurcumin: an orally bioavailable blocker of TNF and other pro-inflammatory biomarkersAggarwal, Bharat B.; Gupta, Subash C.; Sung, BokyungBritish Journal of Pharmacology (2013), 169 (8), 1672-1692CODEN: BJPCBM; ISSN:1476-5381. (Wiley-Blackwell)A review. TNFs are major mediators of inflammation and inflammation-related diseases, hence, the United States Food and Drug Administration (FDA) has approved the use of blockers of the cytokine, TNF-α, for the treatment of osteoarthritis, inflammatory bowel disease, psoriasis and ankylosis. These drugs include the chimeric TNF antibody (infliximab), humanized TNF-α antibody (Humira) and sol. TNF receptor-II (Enbrel) and are assocd. with a total cumulative market value of more than $20 billion a year. As well as being expensive ($15000-20000 per person per yr), these drugs have to be injected and have enough adverse effects to be given a black label warning by the FDA. In the current report, we describe an alternative, curcumin (diferuloylmethane), a component of turmeric (Curcuma longa) that is very inexpensive, orally bioavailable and highly safe in humans, yet can block TNF-α action and prodn. in in vitro models, in animal models and in humans. In addn., we provide evidence for curcumin's activities against all of the diseases for which TNF blockers are currently being used. Mechanisms by which curcumin inhibits the prodn. and the cell signalling pathways activated by this cytokine are also discussed. With health-care costs and safety being major issues today, this golden spice may help provide the soln.
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Abstract
Figure 1
Figure 1. (A) Reaction scheme illustrates the structures of the polymer library and their polymerization conditions. (B) SEC traces of the polymer library using DMAc (0.21 wt % LiCl) as eluent. (C) Titration curves of the polymer library were determined using 0.15 M NaOH as the base with a Metrohm OMNIS integrated titration system.
Figure 2
Figure 2. (A) Experiment design of EBA (first two steps) and HRA (all three steps). (B) EBA was performed at different N*/P ratios ranging from 3 to 20. (C) HRA assay at N*/P = 20. All data points were performed in triplicate, and values were fitted using a B-Spline function. (D) Hydrodynamic size measurement via DLS. (E) Cryo-TEM image of polyplexes of C1.
Figure 3
Figure 3. (A) Mechanism of the cytotoxicity assay (PrestoBlue assay). (B) PrestoBlue assay in L929 over 24 h in a full growth medium (D10H). Dots represent values of single repetitions. Lines were fitted, and IC50 values were calculated with dose–response function (n = 3). Stars indicate the polymer concentration (μg mL–1), which induces 50% cytotoxicity. Viability below 70% was considered cytotoxic. (C) Hemolysis assay was performed in triplicate with three different donors. A relative hemolysis of > 2% is considered slightly hemolytic, and > 5% is considered hemolytic.
Figure 4
Figure 4. (A) Schematic illustration of complexation between the polymers and pDNA and YOYO-1. The experiment was used to determine the particle uptake behavior. (B) Particle uptake was performed in HEK293T cells with full growth medium (D10H) over 1 and 4 h at N*/P ratio 20 and c(pDNA) = 3 μg mL–1 on cells (n = 3). The gating strategy can be found in the Supporting Information, Figure S9.
Figure 5
Figure 5. (A) Schematic illustration of the experimental design for determining transfection efficiency under two different conditions (4 + 20 and 24 h). (B) Transfection efficiency was performed in HEK293T cells with full growth medium (D10H) over 4 + 20 h at N*/P 20 and c(pDNA) = 3 μg mL–1 on cells (n = 3). (C) Performed over 24 h at N*/P 20 and three different concentrations of the genetic material (n = 3). Details of statistical tests can be found in the Supporting Information, Tables S6–S11.
Figure 6
Figure 6. Transfection efficiency was performed in HEK293T cells with full growth medium (D10H) over 24 h at different N*/P ratios and c(pDNA) = 3 μg mL–1 on cells for A1, B2, and C1 (n ≥ 3). Details of statistical tests can be found in the Supporting Information, Tables S12–S15.
Figure 7
Figure 7. (A) Transfection efficiency was performed with A1, B1, B2, and C1 in hard-to-transfect suspension cell lines THP-1 and Jurkat. Transfection was performed in full growth medium (R10H) over 24 h at N*/P 20 and 3 μg mL–1 genetic material on cells (n = 3). (B) Fold change (compared to non-treated cells) of the relative TNF-α mRNA expression (normalized to GAPDH) of ex vivo murine classical monocytes from the bone marrow. One point represents one biological replicate (mice, blue = male, black = female). Mean with standard deviation is depicted. Details of statistical tests can be found in the Supporting Information, Tables S16 and S17.
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- 6Sayed, N.; Allawadhi, P.; Khurana, A.; Singh, V.; Navik, U.; Pasumarthi, S. K.; Khurana, I.; Banothu, A. K.; Weiskirchen, R.; Bharani, K. K. Gene therapy: Comprehensive overview and therapeutic applications. Life Sci. 2022, 294, 120375, DOI: 10.1016/j.lfs.2022.1203756https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BB38XivF2iu74%253D&md5=f76332487e207cdcdeecc77113c5a072Gene therapy: Comprehensive overview and therapeutic applicationsSayed, Nilofer; Allawadhi, Prince; Khurana, Amit; Singh, Vishakha; Navik, Umashanker; Pasumarthi, Sravan Kumar; Khurana, Isha; Banothu, Anil Kumar; Weiskirchen, Ralf; Bharani, Kala KumarLife Sciences (2022), 294 (), 120375CODEN: LIFSAK; ISSN:0024-3205. (Elsevier B.V.)A review. Gene therapy is the product of man's quest to eliminate diseases. Gene therapy has three facets namely, gene silencing using siRNA, shRNA and miRNA, gene replacement where the desired gene in the form of plasmids and viral vectors, are directly administered and finally gene editing based therapy where mutations are modified using specific nucleases such as zinc-finger nucleases (ZFNs), transcription activator-like effector nucleases (TALENs) and clustered regulatory interspaced short tandem repeats (CRISPR)/CRISPR-assocd. protein (Cas)-assocd. nucleases. Transfer of gene is either through transformation where under specific conditions the gene is directly taken up by the bacterial cells, transduction where a bacteriophage is used to transfer the genetic material and lastly transfection that involves forceful delivery of gene using either viral or non-viral vectors. The non-viral transfection methods are subdivided into phys., chem. and biol. The phys. methods include electroporation, biolistic, microinjection, laser, elevated temp., ultrasound and hydrodynamic gene transfer. The chem. methods utilize calcium- phosphate, DAE-dextran, liposomes and nanoparticles for transfection. The biol. methods are increasingly using viruses for gene transfer, these viruses could either integrate within the genome of the host cell conferring a stable gene expression, whereas few other non-integrating viruses are episomal and their expression is dild. proportional to the cell division. So far, gene therapy has been wielded in a plethora of diseases. However, coherent and innocuous delivery of genes is among the major hurdles in the use of this promising therapy. Hence this review aims to highlight the current options available for gene transfer along with the advantages and limitations of every method.
- 7Gaspar, R.; Coelho, F.; Silva, B. F. Lipid-nucleic acid complexes: Physicochemical aspects and prospects for cancer treatment. Molecules 2020, 25 (21), 5006, DOI: 10.3390/molecules252150067https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BB3cXitleit7jJ&md5=a3870e755b37ace60de4b9ed943abc4aLipid-nucleic acid complexes: physicochemical aspects and prospects for cancer treatmentGaspar, Ricardo; Coelho, Filipe; Silva, Bruno F. B.Molecules (2020), 25 (21), 5006CODEN: MOLEFW; ISSN:1420-3049. (MDPI AG)A review. Cancer is an extremely complex disease, typically caused by mutations in cancer-crit. genes. By delivering therapeutic nucleic acids (NAs) to patients, gene therapy offers the possibility to supplement, repair or silence such faulty genes or to stimulate their immune system to fight the disease. While the challenges of gene therapy for cancer are significant, the latter approach (a type of immunotherapy) starts showing promising results in early-stage clin. trials. One important advantage of NA-based cancer therapies over synthetic drugs and protein treatments is the prospect of a more universal approach to designing therapies. Designing NAs with different sequences, for different targets, can be achieved by using the same technologies. This versatility and scalability of NA drug design and prodn. on demand open the way for more efficient, affordable and personalized cancer treatments in the future. However, the delivery of exogenous therapeutic NAs into the patients' targeted cells is also challenging. Membrane-type lipids exhibiting permanent or transient cationic character have been shown to assoc. with NAs (anionic), forming nanosized lipid-NA complexes. These complexes form a wide variety of nanostructures, depending on the global formulation compn. and properties of the lipids and NAs. Importantly, these different lipid-NA nanostructures interact with cells via different mechanisms and their therapeutic potential can be optimized to promising levels in vitro. The complexes are also highly customizable in terms of surface charge and functionalization to allow a wide range of targeting and smart-release properties. Most importantly, these synthetic particles offer possibilities for scaling-up and affordability for the population at large. Hence, the versatility and scalability of these particles seem ideal to accommodate the versatility that NA therapies offer. While in vivo efficiency of lipid-NA complexes is still poor in most cases, the advances achieved in the last three decades are significant and very recently a lipid-based gene therapy medicine was approved for the first time (for treatment of hereditary transthyretin amyloidosis). Although the path to achieve efficient NA-delivery in cancer therapy is still long and tenuous, these advances set a new hope for more treatments in the future. In this review, we attempt to cover the most important biophys. and physicochem. aspects of non-viral lipid-based gene therapy formulations, with a perspective on future cancer treatments in mind.
- 8Ginn, S. L.; Amaya, A. K.; Alexander, I. E.; Edelstein, M.; Abedi, M. R. Gene therapy clinical trials worldwide to 2017: An update. J. Gene Med. 2018, 20 (5), e3015 DOI: 10.1002/jgm.3015There is no corresponding record for this reference.
- 9Dunbar, C. E.; High, K. A.; Joung, J. K.; Kohn, D. B.; Ozawa, K.; Sadelain, M. Gene therapy comes of age. Science 2018, 359 (6372), eaan4672 DOI: 10.1126/science.aan4672There is no corresponding record for this reference.
- 10Shi, B.; Zheng, M.; Tao, W.; Chung, R.; Jin, D.; Ghaffari, D.; Farokhzad, O. C. Challenges in DNA delivery and recent advances in multifunctional polymeric DNA delivery systems. Biomacromolecules 2017, 18 (8), 2231– 2246, DOI: 10.1021/acs.biomac.7b0080310https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC2sXhtVOnsbnL&md5=bff2ce2f4f341f3b182f4da5d3840a61Challenges in DNA Delivery and Recent Advances in a Multifunctional Polymeric DNA Delivery SystemsShi, Bingyang; Zheng, Meng; Tao, Wei; Chung, Roger; Jin, Dayong; Ghaffari, Dariush; Farokhzad, Omid C.Biomacromolecules (2017), 18 (8), 2231-2246CODEN: BOMAF6; ISSN:1525-7797. (American Chemical Society)After more than 20 years of intensive investigations, gene therapy has become one of the most promising strategies for treating genetic diseases. However, the lack of ideal delivery systems has limited the clin. realization of gene therapy's tremendous potential, esp. for DNA-based gene therapy. Over the past decade, considerable advances have been made in the application of polymer-based DNA delivery systems for gene therapy, esp. through multifunctional systems. The core concept behind multifunctional polymeric DNA delivery systems is to endow one single DNA carrier, via materials engineering and surface modification, with several active functions, e.g., good cargo DNA protection, excellent colloidal stability, high cellular uptake efficiency, efficient endo/lysosome escape, effective import into the nucleus, and DNA unpacking. Such specially developed vectors would be capable of overcoming multiple barriers to the successful delivery of DNA. In this review, we first provide a comprehensive overview of the interactions between the protein corona and DNA vectors, the mechanisms and challenges of nonviral DNA vectors, and important concepts in the design of DNA carriers identified via past reports on DNA delivery systems. Finally, we highlight and discuss recent advances in multifunctional polymeric DNA delivery systems based on "off-the-shelf" polycations including polyethylenimine (PEI), poly-L-lysine (PLL), and chitosan and offer perspectives on future developments.
- 11Guo, X.; Huang, L. Recent advances in nonviral vectors for gene delivery. Acc. Chem. Res. 2012, 45 (7), 971– 979, DOI: 10.1021/ar200151m11https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC3MXhtVylsbvK&md5=5b429971bf547460e451ad362d6edcaeRecent Advances in Nonviral Vectors for Gene DeliveryGuo, Xia; Huang, LeafAccounts of Chemical Research (2012), 45 (7), 971-979CODEN: ACHRE4; ISSN:0001-4842. (American Chemical Society)A review. Gene therapy has long been regarded a promising treatment for many diseases, whether acquired (such as AIDS or cancer) or inherited through a genetic disorder. A drug based on a nucleic acid, however, must be delivered to the interior of the target cell while surviving an array of biol. defenses honed by evolution. Successful gene therapy is thus dependent on the development of an efficient delivery vector. Researchers have pursued two major vehicles for gene delivery: viral and nonviral (synthetic) vectors. Although viral vectors currently offer greater efficiency, nonviral vectors, which are typically based on cationic lipids or polymers, are preferred because of safety concerns with viral vectors. So far, nonviral vectors can readily transfect cells in culture, but efficient nanomedicines remain far removed from the clinic. Overcoming the obstacles assocd. with nonviral vectors to improve the delivery efficiency and therapeutic effect of nucleic acids is thus an active area of current research. The difficulties are manifold, including the strong interaction of cationic delivery vehicles with blood components, uptake by the reticuloendothelial system (RES), toxicity, and managing the targeting ability of the carriers with respect to the cells of interest. Modifying the surface with poly(ethylene glycol), i.e., PEGylation, is the predominant method used to reduce the binding of plasma proteins to nonviral vectors and minimize clearance by the RES after i.v. administration. Nanoparticles that are not rapidly cleared from the circulation accumulate in the tumors because of the enhanced permeability and retention effect, and the targeting ligands attached to the distal end of the PEGylated components allow binding to the receptors on the target cell surface. Neutral and anionic liposomes have been also developed for systemic delivery of nucleic acids in exptl. animal models. Other approaches include (i) designing and synthesizing novel cationic lipids and polymers, (ii) chem. coupling the nucleic acid to peptides, targeting ligands, polymers, or environmentally sensitive moieties, and (iii) utilizing inorg. nanoparticles in nucleic acid delivery. Recently, the different classes of nonviral vectors appear to be converging, and the ability to combine features of different classes of nonviral vectors in a single strategy has emerged. With the strengths of several approaches working in concert, more hurdles assocd. with efficient nucleic acid delivery might therefore be overcome. In this Account, we focus on these novel nonviral vectors, which are classified as multifunctional hybrid nucleic acid vectors, novel membrane/core nanoparticles for nucleic acid delivery, and ultrasound-responsive nucleic acid vectors. We highlight systemic delivery studies and consider the future prospects for nucleic acid delivery. A better understanding of the fate of the nanoparticles inside the cell and of the interactions between the parts of hybrid particles should lead to a delivery system suitable for clin. use. We also underscore the value of sustained release of a nucleic acid in this endeavor; making vectors targeted to cells with sustained release in vivo should provide an interesting research challenge.
- 12Hou, X.; Zaks, T.; Langer, R.; Dong, Y. Lipid nanoparticles for mRNA delivery. Nat. Rev. Mater. 2021, 6 (12), 1078– 1094, DOI: 10.1038/s41578-021-00358-012https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BB3MXhslOnsr3N&md5=651ec4d4569c58d0ad34c9215c02fd38Lipid nanoparticles for mRNA deliveryHou, Xucheng; Zaks, Tal; Langer, Robert; Dong, YizhouNature Reviews Materials (2021), 6 (12), 1078-1094CODEN: NRMADL; ISSN:2058-8437. (Nature Portfolio)A review. MRNA (mRNA) has emerged as a new category of therapeutic agent to prevent and treat various diseases. To function in vivo, mRNA requires safe, effective and stable delivery systems that protect the nucleic acid from degrdn. and that allow cellular uptake and mRNA release. Lipid nanoparticles have successfully entered the clinic for the delivery of mRNA; in particular, lipid nanoparticle-mRNA vaccines are now in clin. use against coronavirus disease 2019 (COVID-19), which marks a milestone for mRNA therapeutics. In this Review, we discuss the design of lipid nanoparticles for mRNA delivery and examine physiol. barriers and possible administration routes for lipid nanoparticle-mRNA systems. We then consider key points for the clin. translation of lipid nanoparticle-mRNA formulations, including good manufg. practice, stability, storage and safety, and highlight preclin. and clin. studies of lipid nanoparticle-mRNA therapeutics for infectious diseases, cancer and genetic disorders. Finally, we give an outlook to future possibilities and remaining challenges for this promising technol.
- 13Kumar, R.; Santa Chalarca, C. F.; Bockman, M. R.; Bruggen, C. V.; Grimme, C. J.; Dalal, R. J.; Hanson, M. G.; Hexum, J. K.; Reineke, T. M. Polymeric delivery of therapeutic nucleic acids. Chem. Rev. 2021, 121 (18), 11527– 11652, DOI: 10.1021/acs.chemrev.0c0099713https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BB3MXpvFKgt78%253D&md5=4f4c596405b997bb94c2973d994547e2Polymeric Delivery of Therapeutic Nucleic AcidsKumar, Ramya; Santa Chalarca, Cristiam F.; Bockman, Matthew R.; Bruggen, Craig Van; Grimme, Christian J.; Dalal, Rishad J.; Hanson, Mckenna G.; Hexum, Joseph K.; Reineke, Theresa M.Chemical Reviews (Washington, DC, United States) (2021), 121 (18), 11527-11652CODEN: CHREAY; ISSN:0009-2665. (American Chemical Society)A review. The advent of genome editing has transformed the therapeutic landscape for several debilitating diseases, and the clin. outlook for gene therapeutics has never been more promising. The therapeutic potential of nucleic acids has been limited by a reliance on engineered viral vectors for delivery. Chem. defined polymers can remediate technol., regulatory, and clin. challenges assocd. with viral modes of gene delivery. Because of their scalability, versatility, and exquisite tunability, polymers are ideal biomaterial platforms for delivering nucleic acid payloads efficiently while minimizing immune response and cellular toxicity. While polymeric gene delivery has progressed significantly in the past four decades, clin. translation of polymeric vehicles faces several formidable challenges. The aim of our Account is to illustrate diverse concepts in designing polymeric vectors towards meeting therapeutic goals of in vivo and ex vivo gene therapy. Here, we highlight several classes of polymers employed in gene delivery and summarize the recent work on understanding the contributions of chem. and architectural design parameters. We touch upon characterization methods used to visualize and understand events transpiring at the interfaces between polymer, nucleic acids, and the physiol. environment. We conclude that interdisciplinary approaches and methodologies motivated by fundamental questions are key to designing high-performing polymeric vehicles for gene therapy.
- 14Pack, D. W.; Hoffman, A. S.; Pun, S.; Stayton, P. S. Design and development of polymers for gene delivery. Nat. Rev. Drug Discovery 2005, 4 (7), 581– 593, DOI: 10.1038/nrd177514https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BD2MXmsFGjtbo%253D&md5=ed3dcd52a8b297ee3d0c1cbb7d5ae43bDesign and development of polymers for gene deliveryPack, Daniel W.; Hoffman, Allan S.; Pun, Suzie; Stayton, Patrick S.Nature Reviews Drug Discovery (2005), 4 (7), 581-593CODEN: NRDDAG; ISSN:1474-1776. (Nature Publishing Group)A review. The lack of safe and efficient gene-delivery methods is a limiting obstacle to human gene therapy. Synthetic gene-delivery agents, although safer than recombinant viruses, generally do not possess the required efficacy. In recent years, a variety of effective polymers have been designed specifically for gene delivery, and much has been learned about their structure-function relationships. With the growing understanding of polymer gene-delivery mechanisms and continued efforts of creative polymer chemists, it is likely that polymer-based gene-delivery systems will become an important tool for human gene therapy.
- 15Durymanov, M.; Reineke, J. Non-viral Delivery of Nucleic Acids: Insight Into Mechanisms of Overcoming Intracellular Barriers. Front. Pharmacol 2018, 9, 971, DOI: 10.3389/fphar.2018.0097115https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC1MXjtVartLw%253D&md5=99126dd3217118f202bc8516198b6b79Non-viral delivery of nucleic acids: insight into mechanisms of overcoming intracellular barriersDurymanov, Mikhail; Reineke, JoshuaFrontiers in Pharmacology (2018), 9 (), 971/1-971/15CODEN: FPRHAU; ISSN:1663-9812. (Frontiers Media S.A.)A review. Delivery of genes, including plasmid DNAs, short interfering RNAs (siRNAs), and mRNAs (mRNAs), using artificial non-viral nanotherapeutics is a promising approach in cancer gene therapy. However, multiple physiol. barriers upon systemic administration remain a key challenge in clin. translation of anti-cancer gene therapeutics. Besides extracellular barriers including sequestration of gene delivery nanoparticles from the bloodstream by resident organ-specific macrophages, and their poor extravasation and tissue penetration in tumors, overcoming intracellular barriers is also necessary for successful delivery of nucleic acids. Whereas for RNA delivery the endosomal barrier holds a key importance, transfer of DNA cargo addnl. requires translocation into the nucleus. Better understanding of crossing membrane barriers by nucleic acid nanoformulations is essential to the improvement of current non-viral carriers. This review aims to summarize relevant literature on intracellular trafficking of non-viral nanoparticles and det. key factors toward surmounting intracellular barriers. Moreover, recent data allowed us to propose new interpretations of current hypotheses of endosomal escape mechanisms of nucleic acid nanoformulations.
- 16Jhaveri, A. M.; Torchilin, V. P. Multifunctional polymeric micelles for delivery of drugs and siRNA. Front. Pharmacol 2014, 5, 77, DOI: 10.3389/fphar.2014.0007716https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A280%3ADC%252BC2cflvVKisA%253D%253D&md5=faf83465811984713e2bc64c8e358819Multifunctional polymeric micelles for delivery of drugs and siRNAJhaveri Aditi M; Torchilin Vladimir PFrontiers in pharmacology (2014), 5 (), 77 ISSN:1663-9812.Polymeric micelles, self-assembling nano-constructs of amphiphilic copolymers with a core-shell structure have been used as versatile carriers for delivery of drugs as well as nucleic acids. They have gained immense popularity owing to a host of favorable properties including their capacity to effectively solubilize a variety of poorly soluble pharmaceutical agents, biocompatibility, longevity, high stability in vitro and in vivo and the ability to accumulate in pathological areas with compromised vasculature. Moreover, additional functions can be imparted to these micelles by engineering their surface with various ligands and cell-penetrating moieties to allow for specific targeting and intracellular accumulation, respectively, to load them with contrast agents to confer imaging capabilities, and incorporating stimuli-sensitive groups that allow drug release in response to small changes in the environment. Recently, there has been an increasing trend toward designing polymeric micelles which integrate a number of the above functions into a single carrier to give rise to "smart," multifunctional polymeric micelles. Such multifunctional micelles can be envisaged as key to improving the efficacy of current treatments which have seen a steady increase not only in hydrophobic small molecules, but also in biologics including therapeutic genes, antibodies and small interfering RNA (siRNA). The purpose of this review is to highlight recent advances in the development of multifunctional polymeric micelles specifically for delivery of drugs and siRNA. In spite of the tremendous potential of siRNA, its translation into clinics has been a significant challenge because of physiological barriers to its effective delivery and the lack of safe, effective and clinically suitable vehicles. To that end, we also discuss the potential and suitability of multifunctional polymeric micelles, including lipid-based micelles, as promising vehicles for both siRNA and drugs.
- 17Gardlík, R.; Pálffy, R.; Hodosy, J.; Lukács, J.; Turna, J.; Celec, P. Vectors and delivery systems in gene therapy. Med. Sci. Monit. 2005, 11 (4), RA110There is no corresponding record for this reference.
- 18Perrier, S. 50th Anniversary Perspective: RAFT Polymerization─ A User Guide. Macromolecules 2017, 50 (19), 7433– 7447, DOI: 10.1021/acs.macromol.7b0076718https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC2sXhsFGms7vL&md5=e58928228fbb93e6392106d4abce2a4a50th Anniversary Perspective: RAFT Polymerization-A User GuidePerrier, SebastienMacromolecules (Washington, DC, United States) (2017), 50 (19), 7433-7447CODEN: MAMOBX; ISSN:0024-9297. (American Chemical Society)A review. This Perspective summarizes the features and limitations of reversible addn.-fragmentation chain transfer (RAFT) polymn., highlighting its strengths and weaknesses, as our understanding of the process, from both a mechanistic and an application point of view, has matured over the past 20 years. It is aimed at both experts in the field and newcomers, including undergraduate and postgraduate students, as well as nonexperts in polymn. who are interested in developing their own polymeric structures by exploiting the simple setup of a RAFT polymn.
- 19Gregory, A.; Stenzel, M. H. Complex polymer architectures via RAFT polymerization: From fundamental process to extending the scope using click chemistry and nature’s building blocks. Prog. Polym. Sci. 2012, 37 (1), 38– 105, DOI: 10.1016/j.progpolymsci.2011.08.00419https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC3MXhsVaqt7%252FO&md5=43cfa17e622a10cf00c31388f74d06d0Complex polymer architectures via RAFT polymerization: From fundamental process to extending the scope using click chemistry and nature's building blocksGregory, Andrew; Stenzel, Martina H.Progress in Polymer Science (2012), 37 (1), 38-105CODEN: PRPSB8; ISSN:0079-6700. (Elsevier Ltd.)A review. Reversible addn. fragmentation chain transfer (RAFT) polymn. has made a huge impact in macromol. design. The first block copolymers were described early on, followed by star polymers and then graft polymers. In the last five years, the types of architectures available have become more and more complex. Star and graft polymers now have block structures within their branches, or a range of different branches can be found growing from one core or backbone. Even the synthesis of hyperbranched polymers can be pos. influenced by RAFT polymn., allowing end group control or control over the branching d. The creative combination of RAFT polymn. with other polymn. techniques, such as ATRP or ring-opening polymn., has extended the array of available architectures. In addn., dendrimers were incorporated either as star core or endfunctionalities. A range of synthetic chem. pathways have been utilized and combined with polymer chem., pathways such as click chem.'. These combinations have allowed the creation of novel structures. RAFT processes have been combined with natural polymers and other naturally occurring building blocks, including carbohydrates, polysaccharides, cyclodextrins, proteins and peptides. The result from the intertwining of natural and synthetic materials has resulted in the formation of hybrid biopolymers. Following these developments over the last few years, it is remarkable to see that RAFT polymn. has grown from a lab curiosity to a polymn. tool that is now been used with confidence in material design. Most of the described synthetic procedures in the literature in recent years, which incorporate RAFT polymn., have been undertaken in order to design advanced materials.
- 20Matyjaszewski, K. Atom transfer radical polymerization (ATRP): current status and future perspectives. Macromolecules 2012, 45 (10), 4015– 4039, DOI: 10.1021/ma300171920https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC38XlsVaqs7w%253D&md5=350b580bd1bb46c21ba5dbfd65a6811dAtom Transfer Radical Polymerization (ATRP): Current Status and Future PerspectivesMatyjaszewski, KrzysztofMacromolecules (Washington, DC, United States) (2012), 45 (10), 4015-4039CODEN: MAMOBX; ISSN:0024-9297. (American Chemical Society)A review. Current status and future perspectives in atom transfer radical polymn. (ATRP) are presented. Special emphasis is placed on mechanistic understanding of ATRP, recent synthetic and process development, and new controlled polymer architectures enabled by ATRP. New hybrid materials based on org./inorg. systems and natural/synthetic polymers are presented. Some current and forthcoming applications are described.
- 21Synatschke, C. V.; Schallon, A.; Jérôme, V.; Freitag, R.; Müller, A. H. E. Influence of Polymer Architecture and Molecular Weight of Poly(2-(dimethylamino)ethyl methacrylate) Polycations on Transfection Efficiency and Cell Viability in Gene Delivery. Biomacromolecules 2011, 12 (12), 4247– 4255, DOI: 10.1021/bm201111d21https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC3MXhtlaqtrzL&md5=5d87903451e38527a7384072d747bb1dInfluence of Polymer Architecture and Molecular Weight of Poly(2-(dimethylamino)ethyl methacrylate) Polycations on Transfection Efficiency and Cell Viability in Gene DeliverySynatschke, Christopher V.; Schallon, Anja; Jerome, Valerie; Freitag, Ruth; Mueller, Axel H. E.Biomacromolecules (2011), 12 (12), 4247-4255CODEN: BOMAF6; ISSN:1525-7797. (American Chemical Society)Nonviral gene delivery with the help of polycations has raised considerable interest in the scientific community over the past decades. Herein, we present a systematic study on the influence of the mol. wt. and architecture of poly(2-(dimethylamino)ethyl methacrylate) (PDMAEMA) on the transfection efficiency and the cytotoxicity in CHO-K1 cells. A library of well-defined homopolymers with a linear and star-shaped topol. (3- and 5-arm stars) was synthesized via atom transfer radical polymn. (ATRP). The mol. wts. of the polycations ranged from 16 to 158 kDa. We found that the cytotoxicity at a given mol. wt. decreased with increasing no. of arms. For a successful transfection a min. mol. wt. was necessary, since the polymers with a no.-av. mol. wt., Mn, below 20 kDa showed negligible transfection efficiency at any of the tested polyelectrolyte complex compns. From the combined anal. of cytotoxicity and transfection data, we propose that polymers with a branched architecture and an intermediate mol. wt. are the most promising candidates for efficient gene delivery, since they combine low cytotoxicity with acceptable transfection results.
- 22Burgevin, F.; Hapeshi, A.; Song, J.-I.; Omedes-Pujol, M.; Christie, A.; Lindsay, C.; Perrier, S. Cationic star copolymers obtained by the arm first approach for gene transfection. Polym. Chem. 2023, 14, 3707– 3717, DOI: 10.1039/D3PY00352CThere is no corresponding record for this reference.
- 23Ros, S.; Freitag, J. S.; Smith, D. M.; Stöver, H. D. H. Charge-Shifting Polycations Based on N, N-(dimethylamino) ethyl Acrylate for Improving Cytocompatibility During DNA Delivery. ACS Omega 2020, 5 (16), 9114– 9122, DOI: 10.1021/acsomega.9b0373423https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BB3cXntVyit7k%253D&md5=592a1183564e7d7132577c5477df722aCharge-Shifting Polycations Based on N,N-(dimethylamino)ethyl Acrylate for Improving Cytocompatibility During DNA DeliveryRos, Samantha; Freitag, Jessica S.; Smith, David M.; Stover, Harald D. H.ACS Omega (2020), 5 (16), 9114-9122CODEN: ACSODF; ISSN:2470-1343. (American Chemical Society)Synthetic polycations are studied extensively as DNA delivery agents because of their ease of prodn., good chem. stability, and low cost relative to viral vectors. This report describes the synthesis of charge-shifting polycations based on N,N-(dimethylamino)ethyl acrylate (DMAEA) and 3-aminopropylmethacryamide (APM), called PAD copolymers, and their use for in vitro DNA delivery into HeLa cells. PAD copolymers of varying compns. were prepd. by RAFT polymn. to yield polymers of controlled mol. wts. with low dispersities. Model hydrolysis studies were carried out to assess the rate of charge-shifting of the polycations by loss of the cationic dimethylaminoethanol side chains. They showed redn. in the net cationic charge by about 10-50% depending on compn. after 2 days at pH 7, forming polyampholytes comprising permanent cationic groups, residual DMAEA, as well as anionic acrylic acid groups. HeLa cells exposed for 4 h to PAD copolymers with the greatest charge-shifting ability showed comparable or higher viability at high concns., relative to the noncharge shifting polycations PAPM and polyethyleneimine (PEI) 2 days post-exposure. Cell uptake efficiency of PAD/60bp-Cy3 DNA polyplexes at 2.5:1 N/P ratio was very high (>95%) for all compns., exceeding the uptake efficiency of PEI polyplexes of equiv. compn. These results suggest that these PAD copolymers, and in particular PAD80 contg. 80 mol % DMAEA, have suitable rates of charge-shifting hydrolysis for DNA delivery, as PAD80 showed reduced cytotoxicity at high concns., while still retaining high uptake efficiencies. In addn., the polyampholytes formed during DMAEA hydrolysis in PAD copolymers can offer enhanced long-term cytocompatibility.
- 24Cook, A. B.; Peltier, R.; Hartlieb, M.; Whitfield, R.; Moriceau, G.; Burns, J. A.; Haddleton, D. M.; Perrier, S. Cationic and hydrolysable branched polymers by RAFT for complexation and controlled release of dsRNA. Polym. Chem. 2018, 9 (29), 4025– 4035, DOI: 10.1039/C8PY00804C24https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC1cXht1Kms7rK&md5=8afc458517ca45d84065985be1275725Cationic and hydrolysable branched polymers by RAFT for complexation and controlled release of dsRNACook, Alexander B.; Peltier, Raoul; Hartlieb, Matthias; Whitfield, Richard; Moriceau, Guillaume; Burns, James A.; Haddleton, David M.; Perrier, SebastienPolymer Chemistry (2018), 9 (29), 4025-4035CODEN: PCOHC2; ISSN:1759-9962. (Royal Society of Chemistry)The controlled release of nucleic acids from cationic polymers is an important criteria for the design of gene delivery systems, and can be difficult to achieve due to the persistent pos. charges required to initially complex the nucleic acids. Here, we report the use of highly branched tertiary amine-rich polymers for the complexation and release of dsRNA over a prolonged period of time. Controlled release of dsRNA is obtained via self-catalyzed hydrolysis of the polymer side chains and assocd. change in electrostatic charge. Reversible addn.-fragmentation chain transfer (RAFT) polymn. was utilized to synthesize a series of branched polymers of 2-(dimethylamino)ethyl acrylate (DMAEA), 3-(dimethylamino)propyl acrylate (DMAPA), and 2-(dimethylamino)ethyl methacrylate (DMAEMA) (MW ∼60 000-200 000 g mol-1) and copolymers thereof. The hydrolysis kinetics of all synthesized polymer materials were followed by 1H NMR spectroscopy. Complexation with dsRNA resulted in the formation of polyplex nanoparticles (N/P ratio of 5) with sizes of approx. 400 nm and surface charges of +15 mV. An agarose gel retardation assay showed sustained release of dsRNA from p(DMAEA-co-DMAEMA) for a period of more than 2 wk. Unlike branched PEI commonly used for gene delivery, the majority of these systems showed little toxicity to cells (NIH3T3 fibroblasts). The results point towards pDMAPA and p(DMAEA-co-DMAEMA) being promising polymers for the controlled release of nucleic acids over prolonged periods.
- 25McCool, M.; Senogles, E. The self-catalysed hydrolysis of poly (N, N-dimethylaminoethyl acrylate). Eur. Polym. J. 1989, 25 (7–8), 857– 860, DOI: 10.1016/0014-3057(89)90054-225https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADyaK3cXotVWj&md5=589bac9a9213f93d4e4a76809ed952a4The self-catalyzed hydrolysis of poly(N,N-dimethylaminoethyl acrylate)McCool, M. B.; Senogles, E.European Polymer Journal (1989), 25 (7-8), 857-60CODEN: EUPJAG; ISSN:0014-3057.Poly(N,N-dimethylaminoethyl acrylate) (I) is a weak base in aq. soln. and slowly hydrolyzes at ambient temp.; the reaction was monitored by 1H and 13C NMR and reacted a limiting conversion of ∼60% after 1-2 wk. Poly(N,N-diethylaminoethyl acrylate) reacted similarly and both polymers underwent ester interchange reactions in MeOH. 13C-NMR spectra of I prepd. by radical polymn. showed no evidence for any defect structures.
- 26Truong, N. P.; Jia, Z.; Burges, M.; McMillan, N. A.; Monteiro, M. J. Self-catalyzed degradation of linear cationic poly (2-dimethylaminoethyl acrylate) in water. Biomacromolecules 2011, 12 (5), 1876– 1882, DOI: 10.1021/bm200219e26https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC3MXltFGmt7s%253D&md5=abc5282a81e205e34d102cb73cd886a0Self-catalyzed degradation of linear cationic poly(2-dimethylaminoethyl acrylate) in waterTruong, Nghia P.; Jia, Zhongfan; Burges, Melinda; McMillan, Nigel A. J.; Monteiro, Michael J.Biomacromolecules (2011), 12 (5), 1876-1882CODEN: BOMAF6; ISSN:1525-7797. (American Chemical Society)We report the synthesis of a low cytotoxic polycation that maintains its cationic strength for well over a few hours then degrades into a benign polymer with nontoxic byproducts. Well-defined poly(2-dimethylaminoethyl acrylate) (PDMAEA) of five different mol. wts. prepd. using reversible addn.-fragmentation chain transfer (RAFT) "living" radical polymn. degrades slowly over 200 h (∼8 days). As this degrdn. is independent of both the polymer mol. wt. and soln. pH, it is consistent with a self-catalyzed hydrolysis process without the need for an internal or external degrdn. trigger. In addn., the polymer shows little or no cytotoxicity to HeLa cells for the mol. wts. of 5600 and below, even at very high polymer concns. (equiv. to a nitrogen/phosphorus ratio of 200). Therefore, at sufficiently low mol. wts. this polymer has the essential attributes (i.e., ability to autodegradable and low toxicity) for a delivery carrier suitable for DNA or siRNA.
- 27Truong, N. P.; Jia, Z.; Burgess, M.; Payne, L.; McMillan, N. A.; Monteiro, M. J. Self-catalyzed degradable cationic polymer for release of DNA. Biomacromolecules 2011, 12 (10), 3540– 3548, DOI: 10.1021/bm200742327https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC3MXhtV2rtrbF&md5=640d3cf240d369fc9870395d051c8fd5Self-Catalyzed Degradable Cationic Polymer for Release of DNATruong, Nghia P.; Jia, Zhongfan; Burgess, Melinda; Payne, Liz; McMillan, Nigel A. J.; Monteiro, Michael J.Biomacromolecules (2011), 12 (10), 3540-3548CODEN: BOMAF6; ISSN:1525-7797. (American Chemical Society)The controlled release of siRNA or DNA complexes from cationic polymers is an important parameter design in polymer-based delivery carriers. In this work, we use the self-catalyzed degradable poly(2-dimethylaminoethyl acrylate) (PDMAEA) to strongly bind, protect, and then release oligo DNA (a mimic for siRNA) without the need for a cellular or external trigger. This self-catalyzed hydrolysis process of PDMAEA forms poly(acrylic acid) and N,N'-dimethylamino Et ethanol, both of which have little or no toxicity to cells, and offers the advantage of little or no toxicity to off-target cells and tissues. We found that PDMAEA makes an ideal component of a delivery carrier by protecting the oligo DNA for a sufficiently long period of time to transfect most cells (80% transfection after 4 h) and then has the capacity to release the DNA inside the cells after ∼10 h. The PDMAEA formed large nanoparticle complexes with oligo DNA of ∼400 nm that protected the oligo DNA from DNase in serum. The nanoparticle complexes showed no toxicity for all mol. wts. at a nitrogen/phosphorus (N/P) ratio of 10. Only the higher mol. wt. polymers at very high N/P ratios of 200 showed significant levels of cytotoxicity. These attributes make PDMAEA a promising candidate as a component in the design of a gene delivery carrier without the concern about accumulated toxicity of nanoparticles in the human body after multiadministration, an issue that has become increasingly more important.
- 28Richter, F.; Martin, L.; Leer, K.; Moek, E.; Hausig, F.; Brendel, J. C.; Traeger, A. Tuning of Endosomal Escape and Gene Expression by Functional Groups, Molecular Weight and Transfection Medium: A Structure-Activity Relationship Study. J. Mater. Chem. B 2020, 8, 5026– 5041, DOI: 10.1039/D0TB00340AThere is no corresponding record for this reference.
- 29Richter, F.; Leer, K.; Martin, L.; Mapfumo, P.; Solomun, J. I.; Kuchenbrod, M. T.; Hoeppener, S.; Brendel, J. C.; Traeger, A. The impact of anionic polymers on gene delivery: how composition and assembly help evading the toxicity-efficiency dilemma. J. Nanobiotechnol. 2021, 19, 292, DOI: 10.1186/s12951-021-00994-229https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BB38XivVOnsL4%253D&md5=5ce1c855765214f4b7a0cd5fd455ce00The impact of anionic polymers on gene delivery: how composition and assembly help evading the toxicity-efficiency dilemmaRichter, Friederike; Leer, Katharina; Martin, Liam; Mapfumo, Prosper; Solomun, Jana I.; Kuchenbrod, Maren T.; Hoeppener, Stephanie; Brendel, Johannes C.; Traeger, AnjaJournal of Nanobiotechnology (2021), 19 (1), 292CODEN: JNOAAO; ISSN:1477-3155. (BioMed Central Ltd.)Cationic polymers have been widely studied for non-viral gene delivery due to their ability to bind genetic material and to interact with cellular membranes. However, their charged nature carries the risk of increased cytotoxicity and interaction with serum proteins, limiting their potential in vivo application. Therefore, hydrophilic or anionic shielding polymers are applied to counteract these effects. Herein, a series of micelle-forming and micelle-shielding polymers were synthesized via RAFT polymn. The copolymer poly[(Bu acrylate)-b-(2-(di-Me amino)ethyl acrylamide)] (P(nBA-b-DMAEAm)) was assembled into cationic micelles and different shielding polymers were applied, i.e., poly(acrylic acid) (PAA), poly(4-acryloyl morpholine) (PNAM) or P(NAM-b-AA) block copolymer. These systems were compared to a triblock terpolymer micelle comprising PAA as the middle block. The assemblies were investigated regarding their morphol., interaction with pDNA, cytotoxicity, transfection efficiency, polyplex uptake and endosomal escape. The naked cationic micelle exhibited superior transfection efficiency, but increased cytotoxicity. The addn. of shielding polymers led to reduced toxicity. In particular, the triblock terpolymer micelle convinced with high cell viability and no significant loss in efficiency. The highest shielding effect was achieved by layering micelles with P(NAM-b-AA) supporting the colloidal stability at neutral zeta potential and completely restoring cell viability while maintaining moderate transfection efficiencies. The high potential of this micelle-layer-combination for gene delivery was illustrated for the first time.
- 30Truong, N. P.; Gu, W.; Prasadam, I.; Jia, Z.; Crawford, R.; Xiao, Y.; Monteiro, M. J. An influenza virus-inspired polymer system for the timed release of siRNA. Nat. Commun. 2013, 4 (1), 1902, DOI: 10.1038/ncomms290530https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A280%3ADC%252BC3snlvFCqtw%253D%253D&md5=a9c0ca3205517013c3f753b32a43e482An influenza virus-inspired polymer system for the timed release of siRNATruong Nghia P; Gu Wenyi; Prasadam Indira; Jia Zhongfan; Crawford Ross; Xiao Yin; Monteiro Michael JNature communications (2013), 4 (), 1902 ISSN:.Small interfering RNA silences specific genes by interfering with mRNA translation, and acts to modulate or inhibit specific biological pathways; a therapy that holds great promise in the cure of many diseases. However, the naked small interfering RNA is susceptible to degradation by plasma and tissue nucleases and due to its negative charge unable to cross the cell membrane. Here we report a new polymer carrier designed to mimic the influenza virus escape mechanism from the endosome, followed by a timed release of the small interfering RNA in the cytosol through a self-catalyzed polymer degradation process. Our polymer changes to a negatively charged and non-toxic polymer after the release of small interfering RNA, presenting potential for multiple repeat doses and long-term treatment of diseases.
- 31Tran, N. T.; Truong, N. P.; Gu, W.; Jia, Z.; Cooper, M. A.; Monteiro, M. J. Timed-release polymer nanoparticles. Biomacromolecules 2013, 14 (2), 495– 502, DOI: 10.1021/bm301721k31https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC3sXkt1yguw%253D%253D&md5=f04eb155e4ff9c77681803ac317585bfTimed-Release Polymer NanoparticlesTran, Nguyen T. D.; Truong, Nghia P.; Gu, Wenyi; Jia, Zhongfan; Cooper, Matthew A.; Monteiro, Michael J.Biomacromolecules (2013), 14 (2), 495-502CODEN: BOMAF6; ISSN:1525-7797. (American Chemical Society)Triggered-release of encapsulated therapeutics from nanoparticles without remote or environmental triggers was demonstrated in this work. Disassembly of the polymer nanoparticles to unimers at precise times allowed the controlled release of oligo DNA. The polymers used in this study consisted of a hydrophilic block for stabilization and second thermoresponsive block for self-assembly and disassembly. At temps. below the second block's LCST (i.e., below 37 °C for in vitro assays), the diblock copolymer was fully water-sol., and when heated to 37 °C, the polymer self-assembled into a narrow size distribution of nanoparticles with an av. diam. of approx. 25 nm. The thermoresponsive nature of the second block could be manipulated in situ by the self-catalyzed degrdn. of cationic 2-(dimethylamino)ethyl acrylate (DMAEA) units to neg. charged acrylic acid groups and when the amt. of acid groups was sufficiently high to increase the LCST of the second block above 37 °C. The disassembly of the nanoparticles could be controlled from 10 to 70 h. The use of these nanoparticles as a combined therapy, in which one or more agents can be released in a predetd. way, has the potential to improve the personal point of care treatment of patients.
- 32Tran, N. T.; Jia, Z.; Truong, N. P.; Cooper, M. A.; Monteiro, M. J. Fine tuning the disassembly time of thermoresponsive polymer nanoparticles. Biomacromolecules 2013, 14 (10), 3463– 3471, DOI: 10.1021/bm400785832https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC3sXhsVCgtbbP&md5=e1252dea2d4bd6ec81784d2a3a67cd78Fine Tuning the Disassembly Time of Thermoresponsive Polymer Nanoparticles.Tran, Nguyen T. D.; Jia, Zhongfan; Truong, Nghia P.; Cooper, Matthew A.; Monteiro, Michael J.Biomacromolecules (2013), 14 (10), 3463-3471CODEN: BOMAF6; ISSN:1525-7797. (American Chemical Society)Timed-released disassembly of nanoparticles without a remote trigger or environmental cues is demonstrated in this work. The reversible addn.-fragmentation chain transfer (RAFT) polymn. allowed the fine-tuning of the chem. compn. in the diblock copolymers, in which the first block consisted of a hydrophilic monomer (DMA) and the second random block consisted of three different monomers: (a) the thermoresponsive NIPAM, (b) the self-catalyzed hydrolyzable DMAEA, and (c) the hydrophobic BA. These diblock copolymers were solubilized in water below the lower crit. soln. temp. (LCST) of the thermoresponsive second block, and heated to 37 °C (i.e., >LCST) to form small micelle nanoparticles with a narrow particle size distribution. As DMAEA hydrolyzed to acrylic acid groups, the LCST of the diblock increased, and the time at the start of micelle disassembly (tstart) corresponded to the point where the LCST was equal to the soln. temp. (i.e., 37 °C). The high water content in the PNIPAM core allowed an even degrdn. of the core over time. The copolymer compn. allowed fine control over tstart, as this time was linearly dependent upon the BA units in the second block. These nanoparticles could also be designed to be stable (i.e., not disassemble) over a wide pH range or disassemble below a pH of 7.3. Addnl., the time from the start of disassembly to full unimer formation (tdegrade) could be controlled by the amt. of DMAEA units in the second block. A longer tdegrade (∼5.5 h) was found when the no. of DMAEA units was 42 compared to tdegrade of 1.1 h for 25 units. The nanoparticles designed in this work, through fine control of the polymer chem. compn., have the potential for drug delivery purposes for timed-release of drugs and prodrugs and other wide-ranging applications where timed-release would be beneficial.
- 33Solomun, J. I.; Cinar, G.; Mapfumo, P.; Richter, F.; Moek, E.; Hausig, F.; Martin, L.; Hoeppener, S.; Nischang, I.; Traeger, A. Solely aqueous formulation of hydrophobic cationic polymers for efficient gene delivery. Int. J. Pharm. 2021, 593, 120080, DOI: 10.1016/j.ijpharm.2020.12008033https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BB3cXisFSnsb7I&md5=bc64bd44d33d1a5bf1cc69d27aa4a7a1Solely aqueous formulation of hydrophobic cationic polymers for efficient gene deliverySolomun, Jana I.; Cinar, Gizem; Mapfumo, Prosper; Richter, Friederike; Moek, Elisabeth; Hausig, Franziska; Martin, Liam; Hoeppener, Stephanie; Nischang, Ivo; Traeger, AnjaInternational Journal of Pharmaceutics (Amsterdam, Netherlands) (2021), 593 (), 120080CODEN: IJPHDE; ISSN:0378-5173. (Elsevier B.V.)Cationic polymers are promising gene delivery vectors due to their ability to bind and protect genetic material. The introduction of hydrophobic moieties into cationic polymers can further improve the vector efficiency, but common formulations of hydrophobic polymers involve harsh conditions such as org. solvents, impairing intactness and loading efficiency of the genetic material. In this study, a mild, aq. formulation method for the encapsulation of high amts. of genetic material is presented. A well-defined pH-responsive hydrophobic copolymer, i.e. poly((n-butylmethacrylate)-co-(methylmethacrylate)-co-(2-(dimethylamino) ethylmethacrylate)), (PBMD) was synthesized by reversible addn. fragmentation chain transfer (RAFT) polymn. Exploiting the pH-dependent soly. behavior of the polymer, stable pDNA loaded nanoparticles were prepd. and characterized using anal. ultracentrifugation (AUC), cryo-transmission electron microscopy (cryo-TEM) and dynamic light scattering (DLS). This novel formulation approach showed high transfection efficiencies in HEK293T cells, while requiring 5- to 10-fold less pDNA compared to linear polyethylenimine (LPEI), in particular at short incubation times and in serum-contg. media. Furthermore, the formulation was successfully adopted for siRNA and mRNA encapsulation and the com. approved polymer Eudragit E(PO/100). Overall, the aq. formulation approach, accompanied by a tailor-made hydrophobic polymer and detailed physicochem. and application studies, led to improved gene delivery vectors with high potential for further applications.
- 34Adolph, E. J.; Nelson, C. E.; Werfel, T. A.; Guo, R.; Davidson, J. M.; Guelcher, S. A.; Duvall, C. L. Enhanced Performance of Plasmid DNA Polyplexes Stabilized by a Combination of Core Hydrophobicity and Surface PEGylation. J. Mater. Chem. B 2014, 2 (46), 8154– 8164, DOI: 10.1039/C4TB00352GThere is no corresponding record for this reference.
- 35Convertine, A. J.; Benoit, D. S.; Duvall, C. L.; Hoffman, A. S.; Stayton, P. S. Development of a novel endosomolytic diblock copolymer for siRNA delivery. J. Controlled Release 2009, 133 (3), 221– 229, DOI: 10.1016/j.jconrel.2008.10.00435https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BD1MXpsF2ktQ%253D%253D&md5=e46b2da9dca2388591ffd43dde0bd26bDevelopment of a novel endosomolytic diblock copolymer for siRNA deliveryConvertine, Anthony J.; Benoit, Danielle S. W.; Duvall, Craig L.; Hoffman, Allan S.; Stayton, Patrick S.Journal of Controlled Release (2009), 133 (3), 221-229CODEN: JCREEC; ISSN:0168-3659. (Elsevier B.V.)The gene knockdown activity of small interfering RNA (siRNA) has led to their use as target validation tools and as potential therapeutics for a variety of diseases. The delivery of these double-stranded RNA macromols. has proven to be challenging, however, and in many cases, is a barrier to their deployment. Here we report the development of a new diblock copolymer family that was designed to enhance the systemic and intracellular delivery of siRNA. These diblock copolymers were synthesized using the controlled reversible addn. fragmentation chain transfer polymn. (RAFT) method and are composed of a pos.-charged block of dimethylaminoethyl methacrylate (DMAEMA) to mediate siRNA condensation, and a second endosomal-releasing block composed of DMAEMA and propylacrylic acid (PAA) in roughly equimolar ratios, together with Bu methacrylate (BMA). A related series of diblock compns. were characterized, with the cationic block kept const., and with the ratio of DMAEMA and PAA to BMA varied. These carriers became sharply hemolytic at endosomal pH regimes, with increasing hemolytic activity seen as the percentage of BMA in the second block was systematically increased. The diblock copolymers condensed siRNA into 80-250 nm particles with slightly pos. Zeta potentials. SiRNA-mediated knockdown of a model protein, namely glyceraldehyde 3-phosphate dehydrogenase (GAPDH), in HeLa cells generally followed the hemolytic activity trends, with the most hydrophobic second block (highest BMA content) exhibiting the best knockdown. This pH-responsive carrier designed to mediate endosomal release shows significant promise for the intracellular delivery of siRNA.
- 36Liu, Z.; Zhang, Z.; Zhou, C.; Jiao, Y. Hydrophobic modifications of cationic polymers for gene delivery. Prog. Polym. Sci. 2010, 35 (9), 1144– 1162, DOI: 10.1016/j.progpolymsci.2010.04.00736https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC3cXhtV2isbjI&md5=6ee9ad8ed0fa21a2cce280effb33c3a2Hydrophobic modifications of cationic polymers for gene deliveryLiu, Zonghua; Zhang, Ziyong; Zhou, Changren; Jiao, YanpengProgress in Polymer Science (2010), 35 (9), 1144-1162CODEN: PRPSB8; ISSN:0079-6700. (Elsevier Ltd.)A review. Cationic polymers are the subject of intense research as non-viral gene delivery systems, due to their flexible properties, facile synthesis, robustness, and proven gene delivery efficiency. Nevertheless, low transfection efficiency and undesirable cytotoxicity remain the most challenging aspects of these cationic polymers. To overcome the disadvantages, various modifications were made to improve their gene delivery efficacy. Among them, hydrophobic modifications of the cationic polymers are receiving more and more attention. Most studies have shown that incorporation of hydrophobic chains can improve gene delivery efficiency, mainly explained by hydrophobic interaction conferred to the resulting amphiphilic polycation derivs. and by the enhanced cellular uptake by the hydrophobic chains via the lipophilic cell membrane. This review discusses recent studies on the hydrophobic modifications of cationic polymers for gene delivery. The effects of the hydrophobic modifications are discussed in terms of crit. issues in the gene delivery process, such as gene encapsulation, adsorption to cell membrane, serum inhibition, gene dissocn., cytotoxicity, and tissue-targeting. Moreover, various hydrophobic modifications of the main cationic polymeric gene carriers (polyethylenimine, chitosan, polylysine, etc.) are described with regards to the resulting gene delivery activity. The structure-function relationships discussed here provide important information and insight for the design of novel gene vectors.
- 37Eliyahu, H.; Makovitzki, A.; Azzam, T.; Zlotkin, A.; Joseph, A.; Gazit, D.; Barenholz, Y.; Domb, A. Novel dextran-spermine conjugates as transfecting agents: comparing water-soluble and micellar polymers. Gene Ther. 2005, 12 (6), 494– 503, DOI: 10.1038/sj.gt.330239537https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BD2MXitV2ju7g%253D&md5=fb5a1159e66560b7ecce2f6a5418938cNovel dextran-spermine conjugates as transfecting agents: comparing water-soluble and micellar polymersEliyahu, H.; Makovitzki, A.; Azzam, T.; Zlotkin, A.; Joseph, A.; Gazit, D.; Barenholz, Y.; Domb, A. J.Gene Therapy (2005), 12 (6), 494-503CODEN: GETHEC; ISSN:0969-7128. (Nature Publishing Group)Recently, a novel cationic polymer, dextran-spermine (D-SPM) was developed for gene delivery. An efficient transfection was obtained using this polycation for a variety of genes and cell lines in serum-free or serum-poor medium. However, transfection using the water-sol. D-SPM-based polyplexes decreased with increasing serum concn. in cell culture in a concn.-dependent manner, reaching 95% inhibition at 50% serum in the cell growth medium. In order to overcome this obstacle, oleyl derivs. of D-SPM (which form micelles in aq. phase) were synthesized at 1, 10, and 20 mol% of oleyl moiety to polymer .vepsiln.-NH2 to form N-oleyl-D-SPM (ODS). Polyplexes based on ODS transfected well in medium contg. 50% serum. Comparison with polyplexes based on well-established polymers (branched and linear polyethyleneimine) and with DOTAP/Cholesterol lipoplexes showed that regarding β-galactosidase transgene expression level and cytotoxicity in tissue culture, the D-SPM and ODS compare well with the above polyplexes and lipoplexes. Intracellular trafficking using FITC-labeled ODS and Rhodamine-labeled pGeneGrip plasmid cloned with hBMP2 monitored by confocal microscopy revealed that during the transfection process the fluorescent-labeled polymer concs. in the Golgi app. and around the nucleus, while the cell cytoplasm was free of fluorescent particles, suggesting that the polyplexes move in the cell toward the nucleus by vesicular transport through the cytoplasm and not by a random diffusion. We found that the plasmids penetrate the cell nucleus without the polymer. Preliminary results in zebra fish and mice demonstrate the potential of ODS to serve as an efficient nonviral vector for in vivo transfection.
- 38Mapfumo, P. P.; Reichel, L. S.; Hoeppener, S.; Traeger, A. Improving Gene Delivery: Synergy Between Alkyl Chain Length And Lipoic Acid for Pdmaema Hydrophobic Copolymers. Macromol. Rapid Commun. 2024, 45, 2300649, DOI: 10.1002/marc.202300649There is no corresponding record for this reference.
- 39Mapfumo, P. P.; Solomun, J. I.; Becker, F.; Moek, E.; Leiske, M. N.; Rudolph, L. K.; Brendel, J. C.; Traeger, A. Vitamin B3 Containing Polymers for Nanodelivery. Macromol. Biosci. 2024, 2400002, DOI: 10.1002/mabi.202400002There is no corresponding record for this reference.
- 40Standard, I. 10993-5 Biological Evaluation of Medical Devices. Tests for in Vitro Cytotoxicity; Geneva, Switzerland: International Organization for Standardization, 2009; p 194.There is no corresponding record for this reference.
- 41Leer, K.; Reichel, L. S.; Kimmig, J.; Richter, F.; Hoeppener, S.; Brendel, J. C.; Zechel, S.; Schubert, U. S.; Traeger, A. Optimization of Mixed Micelles Based on Oppositely Charged Block Copolymers by Machine Learning for Application in Gene Delivery. Small 2024, 20, 2306116, DOI: 10.1002/smll.202306116There is no corresponding record for this reference.
- 42Patel, P.; Ibrahim, N. M.; Cheng, K. The importance of apparent pKa in the development of nanoparticles encapsulating siRNA and mRNA. Trends Pharmacol. Sci. 2021, 42 (6), 448– 460, DOI: 10.1016/j.tips.2021.03.00242https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BB3MXns1aqtL0%253D&md5=151ad19fba69fb7f0be6cd37fbbc9376The Importance of Apparent pKa in the Development of Nanoparticles Encapsulating siRNA and mRNAPatel, Pratikkumar; Ibrahim, Nurudeen Mohammed; Cheng, KunTrends in Pharmacological Sciences (2021), 42 (6), 448-460CODEN: TPHSDY; ISSN:0165-6147. (Elsevier Ltd.)A review. Polymer and lipid nanoparticles have been extensively used as carriers to address the biol. barriers encountered in siRNA and mRNA delivery. We summarize the crucial role of nanoparticle charge and ionizability in complexing RNAs, binding to biol. components, escaping from the endosome, and releasing RNAs into the cytoplasm. We highlight the significant impact of the apparent pKa of nanoparticles on their efficacy and toxicity, and the importance of optimizing pKa in the development of lead formulations for RNAs. We also discuss the feasibility of fine-tuning the pKa in nanoparticles and the applications of this approach in the optimization of delivery systems for RNAs.
- 43Alabi, C. A.; Love, K. T.; Sahay, G.; Yin, H.; Luly, K. M.; Langer, R.; Anderson, D. G. Multiparametric approach for the evaluation of lipid nanoparticles for siRNA delivery. Proc. Natl. Acad. Sci. U.S.A. 2013, 110 (32), 12881– 12886, DOI: 10.1073/pnas.130652911043https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC3sXhsVShurbN&md5=f2bc33e036370b72cab58dd701a364b6Multiparametric approach for the evaluation of lipid nanoparticles for siRNA deliveryAlabi, Christopher A.; Love, Kevin T.; Sahay, Gaurav; Yin, Hao; Luly, Kathryn M.; Langer, Robert; Anderson, Daniel G.Proceedings of the National Academy of Sciences of the United States of America (2013), 110 (32), 12881-12886, S12881/1-S12881/22CODEN: PNASA6; ISSN:0027-8424. (National Academy of Sciences)Nanoparticle-mediated siRNA delivery is a complex process that requires transport across numerous extracellular and intracellular barriers. As such, the development of nanoparticles for efficient delivery would benefit from an understanding of how parameters assocd. with these barriers relate to the physicochem. properties of nanoparticles. Here, we use a multiparametric approach for the evaluation of lipid nanoparticles (LNPs) to identify relationships between structure, biol. function, and biol. activity. Our results indicate that evaluation of multiple parameters assocd. with barriers to delivery such as siRNA entrapment, pKa, LNP stability, and cell uptake as a collective may serve as a useful prescreening tool for the advancement of LNPs in vivo. This multiparametric approach complements the use of in vitro efficacy results alone for prescreening and improves in vitro and in vivo translation by minimizing false negatives. For the LNPs used in this work, the evaluation of multiple parameters enabled the identification of LNP pKa as one of the key determinants of LNP function and activity both in vitro and in vivo. It is anticipated that this type of anal. can aid in the identification of meaningful structure-function-activity relationships, improve the in vitro screening process of nanoparticles before in vivo use, and facilitate the future design of potent nanocarriers.
- 44Appleby, C. A.; Wittenberg, B. A.; Wittenberg, J. B. Nicotinic acid as a ligand affecting leghemoglobin structure and oxygen reactivity. Proc. Natl. Acad. Sci. U.S.A. 1973, 70 (2), 564– 568, DOI: 10.1073/pnas.70.2.564There is no corresponding record for this reference.
- 45Ros, S.; Wang, J.; Burke, N. A.; Stöver, H. D. H. A Mechanistic Study of the Hydrolysis of Poly [N, N-(dimethylamino) ethyl acrylates] as Charge-Shifting Polycations. Macromolecules 2020, 53 (9), 3514– 3523, DOI: 10.1021/acs.macromol.9b0227245https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BB3cXntlert7Y%253D&md5=22b718d55109bebc24cc7fcd4426b234A Mechanistic Study of the Hydrolysis of Poly[N,N-(dimethylamino)ethyl acrylates] as Charge-Shifting PolycationsRos, Samantha; Wang, Jiexi; Burke, Nicholas A. D.; Stover, Harald D. H.Macromolecules (Washington, DC, United States) (2020), 53 (9), 3514-3523CODEN: MAMOBX; ISSN:0024-9297. (American Chemical Society)Polycations are used extensively in applications ranging from enhanced oil recovery to biomaterials. Poly[N,N-(dimethylamino)ethyl acrylate] (PDMAEA) has attracted interest for biomaterial applications because of its rapid hydrolysis; however, the mechanism of hydrolysis and the conditions that affect degrdn. often appear amiss in the field. In this report, a detailed 1H NMR spectroscopy study of the hydrolysis of PDMAEA was carried out between pH 0 and 14. In contrast to a widely held view, the rates of hydrolysis of this polymer were found to be highly pH-dependent with half-lives varying from years to minutes as a function of pH. The extent of hydrolysis was also found to be pH-dependent, with a distinct plateau at about 50-60% hydrolysis at pH 7 due to the electrostatic repulsion of anionic carboxylate groups and hydroxide. This was contrasted with the acid-catalyzed mechanism where hydrolysis of PDMAEA at pH 0.3 did not show a plateau in the extent of hydrolysis, reaching 88% hydrolysis after 8 days at 70°C. In addn., the effects of neighboring functional groups on DMAEA hydrolysis in copolymers were explored, with anionic, neutral/hydrophilic, and cationic comonomers found to affect the rates of hydrolysis up to 20-fold at pH 7. Finally, two novel analogs of DMAEA with an addnl. dimethylamino group in different positions of the side chain were synthesized and polymd. to probe the effect of this added tertiary amine on the hydrolysis of the ester linkages. Poly[1,3-bis (dimethylamino)-2-Pr acrylate] (PBDMAPA) hydrolyzed more than 500 times faster at pH 7 than its linear isomer poly[2-((2-(dimethylamino)ethyl)(methyl)amino)ethyl acrylate] (PDEMEA). These results further highlight the importance of the intramol. interactions of the dimethylamino substituent of PDMAEA and the effect of proximity and steric hindrance of the amino group as well as neighboring functional groups of comonomers.
- 46LePecq, J.-B.; Paoletti, C. A fluorescent complex between ethidium bromide and nucleic acids: physical─chemical characterization. J. Mol. Biol. 1967, 27 (1), 87– 106, DOI: 10.1016/0022-2836(67)90353-1There is no corresponding record for this reference.
- 47Olmsted, J., III; Kearns, D. R. Mechanism of ethidium bromide fluorescence enhancement on binding to nucleic acids. Biochemistry 1977, 16 (16), 3647– 3654, DOI: 10.1021/bi00635a02247https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADyaE2sXltFegtLw%253D&md5=7dd2868d98cff0cfb0be2735ca90454bMechanism of ethidium bromide fluorescence enhancement on binding to nucleic acidsOlmsted, John, III; Kearns, David R.Biochemistry (1977), 16 (16), 3647-54CODEN: BICHAW; ISSN:0006-2960.From an examn. of the effect of different solvents on the fluorescence lifetime, quenching of fluorescence by proton acceptors, and the substantial lengthening of lifetime obsd. upon deuteration of the amino protons, regardless of the medium, it appears that proton transfer from the excited singlet state is the process primarily responsible for the low fluorescence yield in most polar solvents. Enhancement of fluorescence upon intercalation is attributed to a redn. in the rate of excited state proton transfer to solvent mols. The proposed mechanism accounts for the ∼3.5-fold increase in the lifetime of free ethidium bromide in going from H2O to D2O; the fact that addn. of small amts. of H2O to nonaq. solvents decreases the fluorescence, whereas addn. of small amts. of D2O enhances the fluorescence; and the enhancement of the ethidium bromide triplet state yield on binding to DNA.
- 48Richter, F.; Mapfumo, P.; Martin, L.; Solomun, J. I.; Hausig, F.; Frietsch, J. J.; Ernst, T.; Hoeppener, S.; Brendel, J. C.; Traeger, A. Improved gene delivery to K-562 leukemia cells by lipoic acid modified block copolymer micelles. J. Nanobiotechnol. 2021, 19 (1), 70, DOI: 10.1186/s12951-021-00801-yThere is no corresponding record for this reference.
- 49Prevette, L. E.; Kodger, T. E.; Reineke, T. M.; Lynch, M. L. Deciphering the role of hydrogen bonding in enhancing pDNA- polycation interactions. Langmuir 2007, 23 (19), 9773– 9784, DOI: 10.1021/la700999549https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BD2sXpt1Wisrk%253D&md5=47380c5628d5e8e7525f1d5c036778a7Deciphering the Role of Hydrogen Bonding in Enhancing pDNA-Polycation InteractionsPrevette, Lisa E.; Kodger, Tom E.; Reineke, Theresa M.; Lynch, Matthew L.Langmuir (2007), 23 (19), 9773-9784CODEN: LANGD5; ISSN:0743-7463. (American Chemical Society)There is considerable interest in the binding and condensation of DNA with polycations to form polyplexes because of their possible application to cellular nucleic acid delivery. This work focuses on studying the binding of plasmid DNA (pDNA) with a series of poly(glycoamidoamine)s (PGAAs) that have previously been shown to deliver pDNA in vitro in an efficient and nontoxic manner. Herein, we examine the PGAA-pDNA binding energetics, binding-linked protonation, and electrostatic contribution to the free energy with isothermal titrn. calorimetry (ITC). The size and charge of the polyplexes at various ITC injection points were then investigated by light scattering and ζ-potential measurements to provide comprehensive insight into the formation of these polyplexes. An anal. of the calorimetric data revealed a three-step process consisting of two different endothermic contributions followed by the condensation/aggregation of polyplexes. The strength of binding and the point of charge neutralization were found to be dependent upon the hydroxyl stereochem. of the carbohydrate moiety within each polymer repeat unit. CD spectra reveal that the PGAAs induce pDNA secondary structure changes upon binding, which suggest a direct interaction between the polymers and the DNA base pairs. IR spectroscopy expts. confirmed both base pair and phosphate group interactions and, more specifically, showed that the stronger-binding PGAAs had more pronounced interactions at both sites. Thus, we conclude that the mechanism of poly(glycoamidoamine)-pDNA binding is most likely a combination of electrostatics and hydrogen bonding in which long-range Coulombic forces initiate the attraction and hydroxyl groups in the carbohydrate comonomer, depending on their stereochem., further enhance the assocn. through hydrogen bonding to the DNA base pairs.
- 50Kong, D. C.; Yang, M. H.; Zhang, X. S.; Du, Z. C.; Fu, Q.; Gao, X. Q.; Gong, J. W. Control of polymer properties by entanglement: a review. Macromol. Mater. Eng. 2021, 306 (12), 2100536, DOI: 10.1002/mame.20210053650https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BB3MXitFans7bJ&md5=d090e9f4b0fe0f536244b08ace880701Control of Polymer Properties by Entanglement: A ReviewKong, De-Chao; Yang, Ming-Hao; Zhang, Xue-Song; Du, Zu-Chen; Fu, Qiang; Gao, Xue-Qin; Gong, Jia-WeiMacromolecular Materials and Engineering (2021), 306 (12), 2100536CODEN: MMENFA; ISSN:1438-7492. (Wiley-VCH Verlag GmbH & Co. KGaA)A review. Chain entanglement, either cohesional or topol., distinguishes polymers from other engineering materials. It impedes the movement of mol. segments and influences the polymer rheol., morphol., and mech. properties. Although a high level of entanglement can increase the polymer toughness, excessive entanglement should be avoided because it causes a high melt viscosity making the processing difficult. This review tended to elucidate the influence of entanglement on the polymer structure, detg. the material properties and processability. A wide range of methods used to fine control the degrees of chain entanglement are summarized. The methods are applicable to polymers in solns., melts, and condensed states with advantages and limitations discussed in detail. The authors also examd. the effect of the entanglement on polymer crystn.-the mechanism remains a controversial issue. This review will provide general guidance to designing and processing polymer materials with desired properties via a rational route of controlling the chain entanglement.
- 51Koltzenburg, S.; Maskos, M.; Nuyken, O.; Mülhaupt, R. Polymere: Synthese, Eigenschaften und Anwendungen; Springer, 2014.There is no corresponding record for this reference.
- 52Rejman, J.; Oberle, V.; Zuhorn, I. S.; Hoekstra, D. Size-dependent internalization of particles via the pathways of clathrin-and caveolae-mediated endocytosis. Biochem. J. 2004, 377 (1), 159– 169, DOI: 10.1042/bj2003125352https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BD2cXis1Wnsg%253D%253D&md5=e517e4c6cbf87ecba1a1c9537a0c67efSize-dependent internalization of particles via the pathways of clathrin- and caveolae-mediated endocytosisRejman, Joanna; Oberle, Volker; Zuhorn, Inge S.; Hoekstra, DickBiochemical Journal (2004), 377 (1), 159-169CODEN: BIJOAK; ISSN:0264-6021. (Portland Press Ltd.)Non-phagocytic eukaryotic cells can internalize particles <1 μm in size, encompassing pathogens, liposomes for drug delivery or lipoplexes applied in gene delivery. In the present study, we have investigated the effect of particle size on the pathway of entry and subsequent intracellular fate in non-phagocytic B16 cells, using a range of fluorescent latex beads of defined sizes (50-1000 nm). Our data reveal that particles as large as 500 nm were internalized by cells via an energy-dependent process. With an increase in size (50-500 nm), cholesterol depletion increased the efficiency of inhibition of uptake. The processing of the smaller particles was significantly perturbed upon microtubule disruption, while displaying a negligible effect on that of the 500 nm beads. Inhibitor and co-localization studies revealed that the mechanism by which the beads were internalized, and their subsequent intracellular routing, was strongly dependent on particle size. Internalization of microspheres with a diam. <200 nm involved clathrin-coated pits. With increasing size, a shift to a mechanism that relied on caveolae-mediated internalization became apparent, which became the predominant pathway of entry for particles of 500 nm in size. At these conditions, delivery to the lysosomes was no longer apparent. The data indicate that the size itself of (ligand-devoid) particles can det. the pathway of entry. The clathrin-mediated pathway of endocytosis shows an upper size limit for internalization of approx. 200 Nm, and kinetic parameters may det. the almost exclusive internalization of such particles along this pathway rather than via caveolae.
- 53Bus, T.; Traeger, A.; Schubert, U. S. The great escape: how cationic polyplexes overcome the endosomal barrier. J. Mater. Chem. B 2018, 6 (43), 6904– 6918, DOI: 10.1039/C8TB00967H53https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC1cXhslSrsLfJ&md5=62d63fe76a4b75545afeaa170bffce43The great escape: how cationic polyplexes overcome the endosomal barrierBus, Tanja; Traeger, Anja; Schubert, Ulrich S.Journal of Materials Chemistry B: Materials for Biology and Medicine (2018), 6 (43), 6904-6918CODEN: JMCBDV; ISSN:2050-7518. (Royal Society of Chemistry)A review. The targeted and efficiency-oriented delivery of (therapeutic) nucleic acids raises hope for successful gene therapy, i.e., for the local and individual treatment of acquired and inherited genetic disorders. Despite promising achievements in the field of polymer-mediated gene delivery, the efficiency of the non-viral vectors remains orders of magnitude lower than viral-mediated ones. Several obstacles on the mol. and cellular level along the gene delivery process were identified, starting from the design and formulation of the nano-sized carriers up to the targeted release to their site of action. In particular, the efficient escape from endo-lysosomal compartments was demonstrated to be a major barrier and its exact mechanism still remains unclear. Different hypotheses and theories of the endosomal escape were postulated. The most popular one is the so-called "proton sponge" hypothesis, claiming an escape by rupture of the endosome through osmotic swelling. It was the first effort to explain the excellent transfection efficiency of poly(ethylene imine). Moreover, it was thought that a unique mechanism based on the ability to capture protons and to buffer the endosomal pH is the basis of endosomal escape. Recent theories deal with the direct interaction of the cationic polyplex or free polymer with the exoplasmic lipid leaflet causing membrane destabilization, permeability or polymer-supported nanoscale hole formation. Both escape strategies are more related to viral-mediated escape compared to the "proton sponge" effect. This review addresses the different endosomal release theories and highlights their key mechanism.
- 54Chen, J.; Wang, K.; Wu, J.; Tian, H.; Chen, X. Polycations for gene delivery: dilemmas and solutions. Bioconj. Chem. 2019, 30 (2), 338– 349, DOI: 10.1021/acs.bioconjchem.8b00688There is no corresponding record for this reference.
- 55Chen, J.; Tian, H.; Dong, X.; Guo, Z.; Jiao, Z.; Li, F.; Kano, A.; Maruyama, A.; Chen, X. Effective Tumor Treatment by VEGF si RNA Complexed with Hydrophobic Poly (A mino Acid)-M odified Polyethylenimine. Macromol. Biosci. 2013, 13 (10), 1438– 1446, DOI: 10.1002/mabi.201300211There is no corresponding record for this reference.
- 56Auwerx, J. The human leukemia cell line, THP-1: a multifacetted model for the study of monocyte-macrophage differentiation. Experientia 1991, 47 (1), 22– 31, DOI: 10.1007/BF0204124456https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A280%3ADyaK3M7ltlCntA%253D%253D&md5=ab4f8ff9f199122d1abcfa85c7b4ee53The human leukemia cell line, THP-1: a multifacetted model for the study of monocyte-macrophage differentiationAuwerx JExperientia (1991), 47 (1), 22-31 ISSN:0014-4754.THP-1 is a human monocytic leukemia cell line. After treatment with phorbol esters, THP-1 cells differentiate into macrophage-like cells which mimic native monocyte-derived macrophages in several respects. Compared to other human myeloid cell lines, such as HL-60, U937, KG-1, or HEL cell lines, differentiated THP-1 cells behave more like native monocyte-derived macrophages. Because of these characteristics, the THP-1 cell line provides a valuable model for studying the mechanisms involved in macrophage differentiation, and for exploring the regulation of macrophage-specific genes as they relate to physiological functions displayed by these cells.
- 57Im, D. J.; Jeong, S. N. Transfection of Jurkat T cells by droplet electroporation. Biochem. Eng. J. 2017, 122, 133– 140, DOI: 10.1016/j.bej.2017.03.01057https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC2sXkvVyrur0%253D&md5=d6e235703f2120a35bf06a9ef4687a1dTransfection of Jurkat T cells by droplet electroporationIm, Do Jin; Jeong, Su-NamBiochemical Engineering Journal (2017), 122 (), 133-140CODEN: BEJOFV; ISSN:1369-703X. (Elsevier B.V.)A droplet electroporation system has been successfully demonstrated for transfection of Jurkat T cell with much higher transfection efficiency (66%) than that of conventional system (11.4%) with the advantages of superior cell viability, comparable throughput, and user-friendly interface. Because the productivity of the proposed system is much greater than previous microfluidic systems, flow cytometry is used for the anal. of transfection efficiency. The high transfection efficiency and cell viability of the droplet electroporation system is mainly attributed to small size which requires lower voltage and provides concd. environment. The successful transfection of Jurkat cell using the droplet electroporation system broadens the applicability of the proposed technol. to medical field. The implication of the present work and the future development direction for a fully automated cell engineering platform are discussed.
- 58Abraham, R. T.; Weiss, A. Jurkat T cells and development of the T-cell receptor signalling paradigm. Nat. Rev. Immunol. 2004, 4 (4), 301– 308, DOI: 10.1038/nri133058https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BD2cXis1GksLY%253D&md5=adc35efbd4d1c9f181ef3c5e262102a7Jurkat T cells and development of the T-cell receptor signalling paradigmAbraham, Robert T.; Weiss, ArthurNature Reviews Immunology (2004), 4 (4), 301-308CODEN: NRIABX; ISSN:1474-1733. (Nature Publishing Group)A review. Twenty years of investigation have yielded a detailed view of the signalling machinery engaged by T-cell receptors (TCRs). Many of the fundamental insights into TCR signalling came from studies carried out with transformed T-cell lines. Perhaps the best known of these model systems is the Jurkat leukemic T-cell line, and here we review some of the key advances in the field of TCR signalling that were made with Jurkat T cells as the host.
- 59Pepe, J.; Rincón, M.; Wu, J. Experimental comparison of sonoporation and electroporation in cell transfection applications. Acoust Res. Lett. Online 2004, 5 (2), 62– 67, DOI: 10.1121/1.1652111There is no corresponding record for this reference.
- 60Schnoor, M.; Buers, I.; Sietmann, A.; Brodde, M. F.; Hofnagel, O.; Robenek, H.; Lorkowski, S. Efficient non-viral transfection of THP-1 cells. J. Immunol. Methods 2009, 344 (2), 109– 115, DOI: 10.1016/j.jim.2009.03.014There is no corresponding record for this reference.
- 61Lipszyc, P. S.; Cremaschi, G. A.; Zubilete, M. Z.; Bertolino, M. L. A.; Capani, F.; Genaro, A. M.; Wald, M. R. Niacin modulates pro-inflammatory cytokine secretion. A potential mechanism involved in its anti-atherosclerotic effect. Open Cardiovasc. Med. J. 2013, 7, 90– 98, DOI: 10.2174/1874192401307010090There is no corresponding record for this reference.
- 62Digby, J. E.; Martinez, F.; Jefferson, A.; Ruparelia, N.; Chai, J.; Wamil, M.; Greaves, D. R.; Choudhury, R. P. Anti-inflammatory effects of nicotinic acid in human monocytes are mediated by GPR109A dependent mechanisms. Atertio. Thromb. Vasc. Biol. 2012, 32 (3), 669– 676, DOI: 10.1161/ATVBAHA.111.241836There is no corresponding record for this reference.
- 63Salem, H. A.; Wadie, W. Effect of niacin on inflammation and angiogenesis in a murine model of ulcerative colitis. Sci. Rep. 2017, 7 (1), 7139, DOI: 10.1038/s41598-017-07280-y63https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A280%3ADC%252BC1cfjs1OmtQ%253D%253D&md5=2985155e9434d34a29446bd3982a8d62Effect of Niacin on Inflammation and Angiogenesis in a Murine Model of Ulcerative ColitisSalem Hesham Aly; Wadie WalaaScientific reports (2017), 7 (1), 7139 ISSN:.Butyrate and niacin are produced by gut microbiota, however butyrate has received most attention for its effects on colonic health. The present study aimed at exploring the effect of niacin on experimental colitis as well as throwing some light on the ability of niacin to modulate angiogenesis which plays a crucial role of in the pathogenesis of inflammatory bowel disease. Rats were given niacin for 2 weeks. On day 8, colitis was induced by intrarectal administration of iodoacetamide. Rats were sacrificed on day 15 and colonic damage was assessed macroscopically and histologically. Colonic myeloperoxidase (MPO), tumour necrosis factor (TNF)-α, interleukin (IL)-10, vascular endothelial growth factor (VEGF), angiostatin and endostatin levels were determined. Niacin attenuated the severity of colitis as demonstrated by a decrease in weight loss, colonic wet weight and MPO activity. Iodoacetamide-induced rise in the colonic levels of TNF-α, VEGF, angiostatin and endostatin was reversed by niacin. Moreover, niacin normalized IL-10 level in colon. Mepenzolate bromide, a GPR109A receptor blocker, abolished the beneficial effects of niacin on body weight, colon wet weight as well as colonic levels of MPO and VEGF. Therefore, niacin was effective against iodoacetamide-induced colitis through ameliorating pathologic angiogenesis and inflammatory changes in a GPR109A-dependent manner.
- 64Si, Y.; Zhang, Y.; Zhao, J.; Guo, S.; Zhai, L.; Yao, S.; Sang, H.; Yang, N.; Song, G.; Gu, J. Niacin inhibits vascular inflammation via downregulating nuclear transcription factor-κB signaling pathway. Mediators Inflamm. 2014, 2014, 1– 12, DOI: 10.1155/2014/263786There is no corresponding record for this reference.
- 65Fitzgerald, K.; O’Neill, L.; Gearing, A.; Callard, R. E. The Cytokine Factsbook and Webfacts; Elsevier, 2001.There is no corresponding record for this reference.
- 66Aggarwal, B. B.; Gupta, S. C.; Sung, B. Curcumin: an orally bioavailable blocker of TNF and other pro-inflammatory biomarkers. Br. J. Pharmacol. 2013, 169 (8), 1672– 1692, DOI: 10.1111/bph.1213166https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC3sXht1elsrnL&md5=3f052e73279fd5eebb2341057dbb7f6bCurcumin: an orally bioavailable blocker of TNF and other pro-inflammatory biomarkersAggarwal, Bharat B.; Gupta, Subash C.; Sung, BokyungBritish Journal of Pharmacology (2013), 169 (8), 1672-1692CODEN: BJPCBM; ISSN:1476-5381. (Wiley-Blackwell)A review. TNFs are major mediators of inflammation and inflammation-related diseases, hence, the United States Food and Drug Administration (FDA) has approved the use of blockers of the cytokine, TNF-α, for the treatment of osteoarthritis, inflammatory bowel disease, psoriasis and ankylosis. These drugs include the chimeric TNF antibody (infliximab), humanized TNF-α antibody (Humira) and sol. TNF receptor-II (Enbrel) and are assocd. with a total cumulative market value of more than $20 billion a year. As well as being expensive ($15000-20000 per person per yr), these drugs have to be injected and have enough adverse effects to be given a black label warning by the FDA. In the current report, we describe an alternative, curcumin (diferuloylmethane), a component of turmeric (Curcuma longa) that is very inexpensive, orally bioavailable and highly safe in humans, yet can block TNF-α action and prodn. in in vitro models, in animal models and in humans. In addn., we provide evidence for curcumin's activities against all of the diseases for which TNF blockers are currently being used. Mechanisms by which curcumin inhibits the prodn. and the cell signalling pathways activated by this cytokine are also discussed. With health-care costs and safety being major issues today, this golden spice may help provide the soln.
Supporting Information
Supporting Information
The Supporting Information is available free of charge at https://pubs.acs.org/doi/10.1021/acs.biomac.4c00007.
Instruments and materials, monomer and polymer synthesis and characterization, polymerization kinetics procedures, titrations, degradation of DMAEA, N*/P ratio calculations, EBA and HRA, cytocompatibility (PrestoBlue and CytoTox-One assay), particle uptake study, transfection efficiency, endosomal escape, antibody mix for murine monocyte staining, and statistics (PDF)
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