# From genotype to phenotype: Molecular and functional characterization of the ID3 SNP rs11574 in CVD

> **NIH NIH F30** · UNIVERSITY OF VIRGINIA · 2020 · $33,965

## Abstract

PROJECT SUMMARY:
 Cardiovascular disease (CVD) and its complications, including myocardial infarction, are the leading
cause of death worldwide. Atherosclerosis, the primary pathological process of CVD, is strongly regulated by
heritable factors, yet the mechanisms and contributions of many of these genetic components to plaque
development are poorly understood. Recently, a single nucleotide polymorphism (SNP) in the coding region of
the gene inhibitor of differentiation 3 (ID3) at rs11574 was identified in several independent studies to be
associated with CVD. Mutation of rs11574 from the major allele (G) to the minor allele (A) changes the 105th
amino acid of ID3 from an alanine to a threonine and is associated with an increased risk of CVD. ID3 functions
as a dominant negative regulator of bHLH transcription factors, and studies in our laboratory show that the minor
allele of rs11574 reduces the ability of ID3 to bind to and sequester E12, increasing E12 occupancy at promoter
regions and enhancing transcription. ID3 regulates vascular smooth muscle cell (VSMC) growth and
differentiation both in vitro and in vivo. VSMCs play an integral role in the stabilization of atherosclerotic lesions
as they contribute to the formation of the fibrous cap within plaques and prevent their rupture. Central to this
process is the ability of VSMCs to proliferate, to regulate inflammation, and to modulate mature VSMC marker
gene expression.
 Deletion of Id3 in Apoe-/- or Ldlr-/- mice significantly increases atherosclerosis, and clinical associations
indicate that rs11574 plays a role in CVD; however, the precise molecular mechanisms through which this coding
SNP alters ID3 biology and CVD risk remain unknown. Studies in this proposal seek to understand the functional
consequences of this ID3 mutant on VSMC biology and will test the hypothesis that the minor allele of rs11574
contributes to detrimental changes in VSMCs in response to atherogenic stimuli. In Aim 1, I propose the creation
of various mutant human induced pluripotent stem cell (iPSC) lines using CRISPR/Cas9 including the full allelic
series of ID3 (A/A, A/G, G/G) as well as knockouts of ID3 and the 2 isoforms of the E2A gene, E12 and E47. I
will differentiate these mutant iPSC lines into VSMCs and compare them for phenotypic differences in
proliferation, inflammation, gene expression, and transcription factor promoter occupancy based upon genotype.
In Aim 2, I will use a VSMC lineage tracing mouse with Id3 specifically deleted only in VSMCs in order to better
understand the contribution of Id3 to atherosclerosis in VSMCs in an in vivo animal model. These Id3-/- mice will
be fed either a Western or control diet for 12 or 24 weeks and will be compared to Id3+/+ mice bred on the same
lineage tracing background in order to quantify differences in atherosclerosis. The proposed studies will enhance
our understanding of the role ID3 and its variants at rs11574 play in atherosclerosis and may identify novel
...

## Key facts

- **NIH application ID:** 10022136
- **Project number:** 5F30HL144116-02
- **Recipient organization:** UNIVERSITY OF VIRGINIA
- **Principal Investigator:** Christopher Andrew Henderson
- **Activity code:** F30 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2020
- **Award amount:** $33,965
- **Award type:** 5
- **Project period:** 2019-07-01 → 2023-06-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10022136

## Citation

> US National Institutes of Health, RePORTER application 10022136, From genotype to phenotype: Molecular and functional characterization of the ID3 SNP rs11574 in CVD (5F30HL144116-02). Retrieved via AI Analytics 2026-05-22 from https://api.ai-analytics.org/grant/nih/10022136. Licensed CC0.

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