Abstract: TH-PO0691
High-Throughput Screening (HTS) to Identify Inhibitors of SH3BP2 in Minimal Change Disease (MCD) and FSGS
Session Information
- Glomerular Diseases: Immunopathogenesis and Targeted Therapeutics
November 06, 2025 | Location: Exhibit Hall, Convention Center
Abstract Time: 10:00 AM - 12:00 PM
Category: Glomerular Diseases
- 1401 Glomerular Diseases: Mechanisms, including Podocyte Biology
Authors
- Srivastava, Tarak, Children's Mercy Kansas City, Kansas City, Missouri, United States
- Roy, Anuradha, University of Kansas, Lawrence, Kansas, United States
- Sharma, Mukut, Kansas City VA Medical Center, Kansas City, Missouri, United States
Background
Increased SH3BP2 expression in human MCD/FSGS (NEPTUNE cohort) glomerular transcriptome corresponds with features of nephrotic syndrome in transgenic mice with a gain-of-function (Sh3bp2KI/KI) mutation (JCI Insight 2024:e170055). We propose that scaffold protein SH3BP2 signaling regulates immune activation in nephrotic syndrome. Current repertoire of chemicals may yield antagonist of SH3BP2.
Methods
Following initial tests, SF8628 Human DIPG H3.3-K27M cells lacking baseline expression of SH3BP2 (Fig. 1a) were selected for lentivirus transduction to deliver GFP (Control) and SH3BP2 gene (Experimental) (Fig 1b). A set of 5019 compounds (1250 FDA-approved drugs and 3769 known bioactives, 5 µM each) were acoustically (ECHO 555) transferred to cells (RT-qPCR validated). After 72h, cells were fixed and nuclei stained with DAPI. Total fluorescence of GFP (λexci 488 nm/ λemi 507 nm) and DAPI (λexci 358/ λemi 461nm) was measured (Agilent Cytation 5, Fig 1c) and percent inhibition of GFP fluorescence calculated using normalized GFP fluorescence.
Results
Initial HTS detected ≥50% GFP inhibition by 257 compounds belonging to 63 mechanism-based clusters. Representative compounds from 43 clusters were confirmed at 0.625, 2.5, and 7.5 µM. These compounds include inhibitors of the mTOR, IκB/IKK and PI3Kα, PI3Kβ, PI3Kδ and PI3Kγ pathways. Cyclosporine, an FDA approved drug identified in this HTS, will be used as a positive control to evaluate candidate molecules in human podocytes.
Conclusion
Current approach may lead to repurpose a drug for modulating SH3BP2-regulated immunopathogenesis in MCD/FSGS.
Figure 1a: The Western Blot showing SH3BP2 expression across 6 different cell lines tested for the study. Figure 1b: The plasmid construct used for lentivirus transduction experiments. Figure 1c: A representative 384 well plate showing GFP and DAPI expression of SF8628 cell line transduced with lentivirus expressing SH3BP2.
Funding
- Other NIH Support