Assays to measure small molecule Hsp70 agonist activity in vitro and in vivo.
Olivia Shapiro, Clara Woods, Amanda M Gleixner, Sara Sannino, Marilyn Ngo, Michael D McDaniels, Peter Wipf, Neil A Hukriede, Christopher J Donnelly, Jeffrey L Brodsky
Author Information
Olivia Shapiro: Department of Neurobiology, LiveLikeLou Center for ALS Research, University of Pittsburgh, Pittsburgh, PA, 15261, USA.
Clara Woods: Department of Cell Biology, Center for Integrative Organ Systems, University of Pittsburgh, Pittsburgh, PA, 15213, USA.
Amanda M Gleixner: Department of Neurobiology, LiveLikeLou Center for ALS Research, University of Pittsburgh, Pittsburgh, PA, 15261, USA.
Sara Sannino: Department of Biological Science, University of Pittsburgh, Pittsburgh, PA, 15260, USA.
Marilyn Ngo: Department of Neurobiology, LiveLikeLou Center for ALS Research, University of Pittsburgh, Pittsburgh, PA, 15261, USA; Department of Human Genetics, University of Pittsburgh School of Public Health, Pittsburgh, PA, 15213, USA.
Michael D McDaniels: Department of Cell Biology, Center for Integrative Organ Systems, University of Pittsburgh, Pittsburgh, PA, 15213, USA.
Peter Wipf: Department of Chemistry, University of Pittsburgh, Pittsburgh, PA, 15260, USA.
Neil A Hukriede: Department of Cell Biology, Center for Integrative Organ Systems, University of Pittsburgh, Pittsburgh, PA, 15213, USA.
Christopher J Donnelly: Department of Neurobiology, LiveLikeLou Center for ALS Research, University of Pittsburgh, Pittsburgh, PA, 15261, USA.
Jeffrey L Brodsky: Department of Biological Science, University of Pittsburgh, Pittsburgh, PA, 15260, USA. Electronic address: jbrodsky@pitt.edu.
Hsp70 prevents protein aggregation and is cytoprotective, but sustained Hsp70 overexpression is problematic. Therefore, we characterized small molecule agonists that augment Hsp70 activity. Because cumbersome assays were required to assay agonists, we developed cell-based and in vivo assays in which disease-associated consequences of Hsp70 activation can be quantified. One assay uses an optogenetic system in which the formation of TDP-43 inclusions can be controlled, and the second assay employs a zebrafish model for acute kidney injury (AKI). These complementary assays will facilitate future work to identify new Hsp70 agonists as well as optimized agonist derivatives.
