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Abstract: PO1658

Novel Small Molecule Compounds Protect Podocytes from Injury In Vitro and In Vivo

Session Information

Category: Glomerular Diseases

  • 1204 Podocyte Biology

Authors

  • Helmuth, Richard, Rush University Medical Center, Chicago, Illinois, United States
  • Noben, Manuel, Rush University Medical Center, Chicago, Illinois, United States
  • Lee, Ha Won, Rush University Medical Center, Chicago, Illinois, United States
  • Saffin, Jean-Michel, Sanford Burnham Prebys Medical Discovery Institute, La Jolla, California, United States
  • Reiser, Jochen, Rush University Medical Center, Chicago, Illinois, United States
  • Heynen-Genel, Susanne, Sanford Burnham Prebys Medical Discovery Institute, La Jolla, California, United States
  • Gupta, Vineet, Rush University Medical Center, Chicago, Illinois, United States

Group or Team Name

  • Gupta lab
Background

Podocyte dysfunction and loss is a key determinant of proteinuria and glomerular injury. Thus, maintaining healthy podocytes is a therapeutic strategy against kidney disease. We previously developed a high-content imaging-based assay and used it to identify a number of small molecule compounds that show protection of podocytes from injury, suggesting it to be a viable strategy for the discovery and development of novel podocyte-protective agents.

Methods

Differentiated mouse podocytes were seeded on collagen-I coated multi-well plates as previously described (Lee et al. JASN, 2015). Cells were exposed to puromycin aminonucleoside (PAN, podocyte injury inducing agent), with compounds from the screening libraries, or DMSO as control, for 48 hours. Cells were fixed and stained which allowed detection using the Opera High-Content Screening (HCS) System. Columbus software was used to quantify morphology properties such as roundness, as well as the overall F-actin signal. Drosophila based screening assays were used to determine efficacy of selected hits in vivo.

Results

PAN damage resulted in quantitative reduction in F-actin fiber numbers and intensity, and increased roundness in podocytes in the ultra-miniaturized assay system. Screening of a library of chemical compounds identified >28 hits that dose-dependently reduced podocyte damage. A set of 9 compounds showed significant protection in a drosophila model of kidney injury, supporting the findings from the high-throughput screening assay.

Conclusion

Our 1536-well plate-based assay system identified a number of small molecule compounds that dose dependently protected podocytes from damage in vitro. A drosophila model of kidney injury validated most of the in vitro data from the podocyte screening assay. Current in vitro and in vivo mechanistic studies are underway to elucidate new insights into podocyte pathways that are therapeutically targeted by the selected hits. These agents hold promise as novel therapeutics for kidney disease patients with podocyte pathologies.

Funding

  • NIDDK Support