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Single Cell MALDI-MSI Analysis of Lipids and Proteins within a Replicative Senescence Fibroblast Model
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    Single Cell MALDI-MSI Analysis of Lipids and Proteins within a Replicative Senescence Fibroblast Model
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    • Emily R. Sekera
      Emily R. Sekera
      Department of Chemistry and Biochemistry, The Ohio State University, Columbus, Ohio 43210 United States
    • Lorena Rosas
      Lorena Rosas
      Department of Internal Medicine, Division of Pulmonary, Critical Care and Sleep Medicine, The Ohio State University, Columbus, Ohio 43210 United States
      More by Lorena Rosas
    • Joseph H. Holbrook
      Joseph H. Holbrook
      Department of Chemistry and Biochemistry, The Ohio State University, Columbus, Ohio 43210 United States
    • Quetzalli D. Angeles-Lopez
      Quetzalli D. Angeles-Lopez
      Department of Internal Medicine, Division of Pulmonary, Critical Care and Sleep Medicine, The Ohio State University, Columbus, Ohio 43210 United States
    • Timur Khaliullin
      Timur Khaliullin
      Department of Internal Medicine, Division of Pulmonary, Critical Care and Sleep Medicine, The Ohio State University, Columbus, Ohio 43210 United States
    • Mauricio Rojas
      Mauricio Rojas
      Department of Internal Medicine, Division of Pulmonary, Critical Care and Sleep Medicine, The Ohio State University, Columbus, Ohio 43210 United States
    • Ana L. Mora
      Ana L. Mora
      Department of Internal Medicine, Division of Pulmonary, Critical Care and Sleep Medicine, The Ohio State University, Columbus, Ohio 43210 United States
      More by Ana L. Mora
    • Amanda B. Hummon*
      Amanda B. Hummon
      Department of Chemistry and Biochemistry, The Ohio State University, Columbus, Ohio 43210 United States
      *Email: [email protected]
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    Journal of the American Society for Mass Spectrometry

    Cite this: J. Am. Soc. Mass Spectrom. 2024, 35, 12, 2815–2823
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    https://doi.org/10.1021/jasms.4c00095
    Published October 30, 2024
    Copyright © 2024 American Society for Mass Spectrometry. Published by American Chemical Society. All rights reserved.

    Abstract

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    In this study, we evaluate lipids and select proteins in human lung fibroblasts (hLFs) to interrogate changes occurring due to aging and senescence. To study single cell populations, a comparison of cells adhered onto slides using poly-d-lysine versus centrifugal force deposition was first analyzed to determine whether specific alterations were observed between preparations. The poly-d-lysine approach was then utilized to interrogate the lipidome of the cell populations and further evaluate potential applications of the MALDI-immunohistochemistry (IHC) platform for single-cell-level analyses. Two protein markers of senescence, vimentin and p21, were both observed within the fibroblast populations and quantified. Lipidomic analysis of the fibroblasts found 12 lipids significantly altered because of replicative senescence, including fatty acids, such as stearic acid, and ceramide phosphoethanolamine species (CerPE). Similar to previous reports, alterations were detected in putative fatty acid building blocks, ceramides, among other lipid species. Altogether, our results reveal the ability to detect lipids implicated in senescence and show alterations to protein expression between normal and senescent fibroblast populations, including differences between young and aged cells. This report is the first time that the MALDI-IHC system has been utilized at a single-cell level to analyze both protein expression and lipid profiles in cultured cells, with a particular focus on changes associated with aging and senescence.

    Copyright © 2024 American Society for Mass Spectrometry. Published by American Chemical Society. All rights reserved.

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    Supporting Information

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    The Supporting Information is available free of charge at https://pubs.acs.org/doi/10.1021/jasms.4c00095.

    • Additional experimental details and MSI images, descriptions of segmentation analyses, fibroblast characterization through Western blot analyses and SA-β-gal assays, and tables detailing the statistical significance determined for interrogated lipid species (PDF)

    • MS peaks (XLSX)

    • NEDC analysis (XLSX)

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    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.

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    Journal of the American Society for Mass Spectrometry

    Cite this: J. Am. Soc. Mass Spectrom. 2024, 35, 12, 2815–2823
    Click to copy citationCitation copied!
    https://doi.org/10.1021/jasms.4c00095
    Published October 30, 2024
    Copyright © 2024 American Society for Mass Spectrometry. Published by American Chemical Society. All rights reserved.

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