# The role of SHROOM3 in congenital heart disease

> **NIH NIH P01** · INDIANA UNIVERSITY INDIANAPOLIS · 2024 · $356,625

## Abstract

ABSTRACT
Congenital heart disease (CHD) is the most common birth defect, yet most etiologies remain unknown.
SHROOM3 is a novel CHD candidate gene that functions through interactions with F-ACTIN, Rho Associated
Coiled-Coil Containing Protein Kinase (ROCK) and Disheveled2 (DVL2) and thus participates in the
noncanonical WNT/Planar Cell Polarity (PCP) signaling pathway. Disruption of PCP signaling leads to cardiac
defects, including left-right patterning defects, left ventricle noncompaction (LVNC), ventricular septal defects
(VSDs) and outflow tract (OFT) defects. However, SHROOM3’s precise role in the PCP signaling cascade, and
in cardiac development, is only partially understood. For example, the Ware laboratory recently identified that
SHROOM3 variants are associated with the left-right patterning defect heterotaxy and CHD in patients. This
finding prompted me to study the impact of the loss of SHROOM3 expression on cardiac development. Utilizing
a gene trap mouse model (Shroom3gt/gt), I demonstrated that SHROOM3 loss-of-function results in CHDs in
embryonic hearts, including VSDs, LVNC and OFT defects in homozygous mutant animals. We also showed
evidence there may be genetic interaction between Shroom3 and Dvl2 during cardiac development. Moreover,
molecular studies have established a protein-protein interaction (either direct or through a common binding
partner) between SHROOM3 and DVL2, though the binding site is poorly defined. Interestingly, a recent study
revealed that canonical WNT/PCP signaling is controlled in part through the deubiquination of DVL2 via USPX9,
a highly conserved deubiquitylase. Utilizing immunoprecipitation (IP)/mass spectrometry (MS) of SHROOM3-
transfected Cos7 cells, I have shown protein-protein interaction between SHROOM3 and USPX9. These
observations have collectively led to the hypothesis that signaling between SHROOM3, DVL2 and USP9X is
necessary for normal cardiac development and that disruption of this signaling can contribute to CHD. There is
evidence SHROOM3 interacts with USP9x and DVL2 however the interactions are at best poorly defined.
Genetic interactions between Shroom3 and Dvl2 during cardiac development have not been not fully delineated
and genetic interaction between Shroom3 and Usp9x are unexplored. Aim 1 will assay genetic interaction
between Shroom3, Dvl2 and Usp9x, analyzing cardiac defects and disrupted PCP endpoints. Although data
demonstrate SHROOM3 binds DVL2, and USP9X, the binding site for DVL2 is only roughly localized to a 490
amino acid region within SHROOM3 and SHROOM3’s USP9X binding site is completely unknown. Aim 2 will
assay protein-protein interactions between SHROOM3, USP9X and DVL2. Defining SHROOM3’s interaction
partners will help determine its role in PCP, cardiac development and CHD pathogenesis. The plan will also build
skills, under close mentorship as an Early Career Investigator, in molecular biology, mouse disease models and
genomics analysis, providing a skillset to i...

## Key facts

- **NIH application ID:** 10755296
- **Project number:** 5P01HL134599-07
- **Recipient organization:** INDIANA UNIVERSITY INDIANAPOLIS
- **Principal Investigator:** Matthew D. Durbin
- **Activity code:** P01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2024
- **Award amount:** $356,625
- **Award type:** 5
- **Project period:** 2017-02-15 → 2028-01-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10755296

## Citation

> US National Institutes of Health, RePORTER application 10755296, The role of SHROOM3 in congenital heart disease (5P01HL134599-07). Retrieved via AI Analytics 2026-05-23 from https://api.ai-analytics.org/grant/nih/10755296. Licensed CC0.

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