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Deconvoluting Stress-Responsive Proteostasis Signaling Pathways for Pharmacologic Activation Using Targeted RNA Sequencing

  • Julia M. D. Grandjean
    Julia M. D. Grandjean
    Department of Molecular Medicine, The Scripps Research Institute, La Jolla, California 92037, United States
  • Lars Plate
    Lars Plate
    Department of Molecular Medicine  and  Department of Chemistry, The Scripps Research Institute, La Jolla, California 92037, United States
    More by Lars Plate
  • Richard I. Morimoto
    Richard I. Morimoto
    Department of Molecular Biosciences, Rice Institute for Biomedical Research, Northwestern University, Evanston, Illinois 60208, United States
  • Michael J. Bollong
    Michael J. Bollong
    Department of Chemistry, The Scripps Research Institute, La Jolla, California 92037, United States
  • Evan T. Powers
    Evan T. Powers
    Department of Chemistry, The Scripps Research Institute, La Jolla, California 92037, United States
  • , and 
  • R. Luke Wiseman*
    R. Luke Wiseman
    Department of Molecular Medicine, The Scripps Research Institute, La Jolla, California 92037, United States
    *Department of Molecular Medicine, The Scripps Research Institute, La Jolla, CA 92037. E-mail: [email protected]. Phone: (858) 784-8820.
Cite this: ACS Chem. Biol. 2019, 14, 4, 784–795
Publication Date (Web):March 1, 2019
Copyright © 2019 American Chemical Society

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    Abstract Image

    Cellular proteostasis is maintained by stress-responsive signaling pathways such as the heat shock response (HSR), the oxidative stress response (OSR), and the unfolded protein response (UPR). Activation of these pathways results in the transcriptional upregulation of select subsets of stress-responsive genes that restore proteostasis and adapt cellular physiology to promote recovery following various types of acute insult. The capacity for these pathways to regulate cellular proteostasis makes them attractive therapeutic targets for correcting proteostasis defects associated with diverse diseases. High-throughput screening (HTS) using cell-based reporter assays is highly effective for identifying putative activators of stress-responsive signaling pathways. However, the development of these compounds is hampered by the lack of medium-throughput assays to define compound potency and selectivity for a given pathway. Here, we describe a targeted RNA sequencing (RNAseq) assay that allows cost-effective, medium-throughput screening of stress-responsive signaling pathway activation. We demonstrate that this assay allows deconvolution of stress-responsive signaling activated by chemical genetic or pharmacologic agents. Furthermore, we use this assay to define the selectivity of putative OSR and HSR activating compounds previously identified by HTS. Our results demonstrate the potential for integrating this adaptable targeted RNAseq assay into screening programs focused on developing pharmacologic activators of stress-responsive signaling pathways.

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