# Cerebral organoid models of CHD2-associated intellectual and developmental disabilities

> **NIH NIH F32** · NORTHWESTERN UNIVERSITY · 2021 · $77,386

## Abstract

PROJECT SUMMARY
Pathogenic variants in CHD2 are associated with intellectual and developmental disabilities (IDDs), including
autism spectrum disorder, developmental delay, and early onset epilepsy. Chromodomain helicase DNA
binding protein 2 (CHD2) is a chromatin remodeling protein that alters chromatin structure, leading to changes
in gene expression of downstream targets. Little is known about how variants in CHD2 lead to IDDs. The
mouse model of Chd2 deficiency does not recapitulate the seizure phenotype seen in human patients with
CHD2 variants, indicating that there may be species-specific functions of CHD2 in the brain. I hypothesize that
CHD2 will be important for regulating genes involved in the development of primate-specific brain structures,
such as the outer subventricular zone. To study the mechanism of CHD2 in a human neuronal system, we
propose the development of a 3D cerebral organoid (CO) model of CHD2 deficiency. Using CRISPR/Cas9, we
have genetically engineered an induced pluripotent stem cell (iPSC) line carrying an endogenous form of
CHD2 with Flag, HA, and green fluorescent protein (GFP) tags (WTCHD2-Flag-HA-GFP). In our first aim, we propose
deletion of one copy of CHD2 using CRISPR/Cas9, resulting in iPSCs haploinsufficient for CHD2 (CHD2+/-Flag-
HA-GFP). WTCHD2-Flag-HA-GFP iPSCs and CHD2+/-Flag-HA-GFP iPSCs will then be differentiated into forebrain specific
COs. I will determine neurodevelopmental defects upon CHD2 disruption, such as differences in neuronal
differentiation, proliferation, and formation of distinct cerebral cortex layers, through a combination of
immunohistochemistry and cell cycle analyses. In Aim 2 I will identify genes misregulated upon CHD2
disruption, by performing single-cell mRNA sequencing in WTCHD2-Flag-HA-GFP and CHD2+/-Flag-HA-GFP COs. In Aim
3, I will determine genome-wide CHD2 occupancy, by performing chromatin immunoprecipitation followed by
next generation sequencing (ChIP-Seq) on cells dissociated from WTCHD2-Flag-HA-GFP COs. I will then correlate
the results of CHD2 occupancy with the data on which genes are misregulated upon CHD2 disruption to
identify a set of candidate genes that are regulated by CHD2 in human neuronal cells. Combined, these
studies will lead to identification of CHD2-regulated genes in human neuronal cell types, revealing pathways in
which CHD2 is involved and improving our understanding of the biological mechanisms behind CHD2-
associated IDDs. In the future this model has the potential for drug development by attempting to rescue
defects with small molecules.

## Key facts

- **NIH application ID:** 10287481
- **Project number:** 5F32HD101280-02
- **Recipient organization:** NORTHWESTERN UNIVERSITY
- **Principal Investigator:** Kay-Marie Lamar
- **Activity code:** F32 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2021
- **Award amount:** $77,386
- **Award type:** 5
- **Project period:** 2020-07-01 → 2023-06-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10287481

## Citation

> US National Institutes of Health, RePORTER application 10287481, Cerebral organoid models of CHD2-associated intellectual and developmental disabilities (5F32HD101280-02). Retrieved via AI Analytics 2026-05-23 from https://api.ai-analytics.org/grant/nih/10287481. Licensed CC0.

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