# Molecular characterization of a novel cause of human FSGS

> **NIH NIH R01** · YALE UNIVERSITY · 2020 · $490,056

## Abstract

Focal segmental glomerulosclerosis (FSGS) is a progressive proteinuric kidney disease and the third
leading cause of end-stage renal disease in the US. Genetic studies demonstrate that FSGS is largely a
podocyte disease. The molecular cause of FSGS is unknown in over half the cases and effective
treatments remain elusive. We performed whole exome sequencing (WES) in two siblings with FSGS and
identified a shared nonsense mutation in MYO9A, which encodes an unconventional myosin not
previously associated with renal disease. This proposal overall objective is to determine whether human
MYO9A mutations cause FSGS, define the molecular pathogenesis and identify novel therapeutic
targets.
MYO9A is a motorized Rho-GAP that controls collective cell migration and negatively regulates RhoA
activity at cell-cell junctions. RhoA modulates podocyte actin dynamics and is important to maintain
glomerular permeabllity, but podocyte RhoA constitutive activation results in proteinuria and
glomerulosclerosis in mice. Preliminary data show that MYO9A is expressed in kidney podocytes and
regulates their size. The MYO9A mutation identified in FSGS patients predicts a truncated gene product
that lacks the RhoA-GAP domain. To assess the functional consequences of this mutation in vivo we
generated Myo9A mutant mice using CRISPR-Cas9 mediated gene editing, which developed proteinuria,
foot process effacement and glomerulosclerosis. We hypothesize that MYO9A mutations can cause
FSGS by disrupting MYO9A podocyte Rho-GAP activity. We propose to evaluate the mutant MYO9A
function in vivo and in vitro, and to identify additional MYO9A mutations in large FSGS/ SRNS cohorts.
Specific aims are: 1) Establish whether MYO9A mutation leads to FSGS/NS in mice by completing the
phenotypic analysis of mutant Myo9A mice and testing whether Rho inhibition modulates the mutant
Myo9A phenotype and renal disease progression. 2) Elucidate the molecular pathogenic mechanism of
the novel FSGS-associated MYO9A mutation by evaluating MYO9A signaling and function in normal
human podocytes vs. MYO9A knockdown and mutant MYO9A podocytes using biochemical and cell
assays. 3) Identify MYO9A mutations in FSGS-Clinical trial patients by WES, and identify additional
patients with MYO9A mutations in large SRNS/FSGS cohorts. These studies will advance our
mechanistic understanding of human FSGS through cells and mice carrying the novel MYO9A mutation,
generated by gene editing. This translational strategy holds promise to deliver personalized,
mechanistically based therapies for glomerular diseases within relatively short time span.

## Key facts

- **NIH application ID:** 9849275
- **Project number:** 5R01DK109434-04
- **Recipient organization:** YALE UNIVERSITY
- **Principal Investigator:** Alda Tufro
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2020
- **Award amount:** $490,056
- **Award type:** 5
- **Project period:** 2017-02-01 → 2022-01-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/9849275

## Citation

> US National Institutes of Health, RePORTER application 9849275, Molecular characterization of a novel cause of human FSGS (5R01DK109434-04). Retrieved via AI Analytics 2026-05-24 from https://api.ai-analytics.org/grant/nih/9849275. Licensed CC0.

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