# Mouse functional analysis of genes for congenital heart disease

> **NIH NIH P01** · ALBERT EINSTEIN COLLEGE OF MEDICINE · 2020 · $374,454

## Abstract

ABSTRACT – PROJECT 3 
This P01 renewal application is in response to RFA, HD-16-009 from the NICHD Institute of NIH. The 
scientific goal for Project 3 is to determine the developmental mechanisms of congenital heart disease using 
the mouse as a model system. The class of congenital heart disease that is the focus of the P01 is termed, 
conotruncal and related aortic arch defects, referred to as CTRDs. As part of the P01, we will identify genes 
and noncoding regions of interest from patients with 22q11.2 deletion syndrome (22q11.2DS; aka DiGeorge 
syndrome velo-cardio-facial syndrome; Project 1) and non-syndromic disease. Among the 40 genes deleted 
in patients with 22q11.2DS, two of them, TBX1, encoding a T-box transcription factor and CRKL, encoding a 
cytoplasmic adaptor for intracellular signaling, are considered the strongest candidate genes. During the 
current P01, we created and utilized conditional loss of function mutant mouse models for Tbx1 and Crkl to 
understand their function. The cardiac outflow tract and aortic arch develop from progenitor anterior heart 
field and neural crest cells that lie within the embryonic pharyngeal apparatus. The Tbx1 gene is expressed 
in the epithelia (endoderm and ectoderm) as well as anterior heart field mesoderm of the pharyngeal 
apparatus, while Crkl is ubiquitously expressed, with highest expression in adjacent neural crest cells. Tbx1 
is not expressed in the developing heart, implicating its function truly in the migrating cells entering the 
heart. Previous studies demonstrated that Tbx1 and Crkl genetically interact, but the mechanistic basis of 
this has only been partially explored. In Aim 1 of this renewal program, we will understand the tissue 
specific mechanism by which Tbx1 and Crkl interact in forming and remodeling the pharyngeal arch arteries. 
In Aim 2, we will perform RNA-seq on the microdissected distal pharyngeal apparatus in wildtype and 
mutant mouse embryos (Tbx1, Crkl and Lgdel/+) and create an interactive gene network termed the PA-INet 
to provide new insights to cardiovascular development and to send to Projects 1 and 2. In Aim 3, we will 
uncover regions of open chromatin from the pharyngeal tissue, so as to prioritize loci harboring noncoding 
variants from SNP genotyping arrays and whole genome sequencing. In addition, Aim 3 will utilize a 
pipeline to perform functional analysis of genes and loci found in Projects 1 and 2, starting with 
computational in silico analysis, in vitro or cell culture assays and finally testing in mouse models. Overall, 
Project 3 is designed to advance the science by determining developmental mechanisms and to enhance 
discoveries made in Projects 1 and 2. 
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## Key facts

- **NIH application ID:** 9938648
- **Project number:** 5P01HD070454-12
- **Recipient organization:** ALBERT EINSTEIN COLLEGE OF MEDICINE
- **Principal Investigator:** BERNICE E MORROW
- **Activity code:** P01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2020
- **Award amount:** $374,454
- **Award type:** 5
- **Project period:** 2011-09-24 → 2022-06-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/9938648

## Citation

> US National Institutes of Health, RePORTER application 9938648, Mouse functional analysis of genes for congenital heart disease (5P01HD070454-12). Retrieved via AI Analytics 2026-05-26 from https://api.ai-analytics.org/grant/nih/9938648. Licensed CC0.

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