# Using human iPSC-derived microglia to understand the biological function of Alzheimer's risk variants

> **NIH NIH F32** · ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI · 2022 · $67,582

## Abstract

Project Summary
Alzheimer’s disease is the most common form of dementia in the elderly, but the causal mechanisms involved
in disease development remain poorly understood. Genome-wide association studies have identified several
genomic regions associated with disease, but translating these into causal variants and genes remains a
challenge. Initial studies have identified an enrichment for AD risk variants in active enhancers of human
monocytes, macrophages, and microglia, suggesting many of these variants act by disrupting gene expression
specifically in myeloid cells. This, along with myeloid-specific expression of several genes implicated in AD
risk, strongly implicate these cells in the etiology of AD. Using fine mapping approaches and integrated
functional data from human myeloid cells, we identified the gene embryonic ectoderm development (EED) as a
strong candidate causal gene and a putative target of a myeloid cell enhancer containing an AD-associated
functional variant on chromosome 11. While EED is known to function in the maintenance and placement of
repressive histone mark H3K27me3 and has been implicated in the regulation of clearance behavior in mouse
microglia, it remains relatively unstudied in the context of AD and its role in human microglia is unclear. The
overall goal of this proposal is to directly test the hypothesis that EED plays a critical role in regulating human
microglia behavior in vitro and in vivo, and to gain a better understanding of how AD-associated genomic
elements may influence EED expression. In Aim 1 I will combine CRISPR gene editing in human induced
pluripotent stem cells (iPSCs), microglial differentiation protocols, novel transplantation methods involving
direct injection of microglia precursor cells into the mouse brain, and functional genomics techniques including
single-nuclei RNA sequencing with targeted functional assays to evaluate the role of EED in human microglia.
Aim 2 will evaluate the mechanism of action of our prioritized variant/enhancer pair and investigate the extent
to which EED is the main target of these AD-associated elements. Specifically, I will use CRISPR to
simultaneously delete the candidate enhancer and create isogenic lines homozygous for the major and minor
alleles of the candidate variant and measure downstream changes in gene expression, chromatin accessibility,
and transcription factor binding. Together, results here will confirm or refute computational predictions that our
prioritized AD-associated variant impacts myeloid cell physiology by altering EED levels in human microglia
cells via a cell-type specific enhancer. This work has the potential to both greatly impact public health through
the validation of a novel AD risk gene and enhance our understanding of genes, pathways, and epigenetic
mechanisms involved in disease development. In addition, functional validation of computational predictions
derived from fine mapping will provide a blueprint for future researchers i...

## Key facts

- **NIH application ID:** 10470313
- **Project number:** 5F32AG069503-03
- **Recipient organization:** ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
- **Principal Investigator:** Sarah M Neuner
- **Activity code:** F32 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2022
- **Award amount:** $67,582
- **Award type:** 5
- **Project period:** 2020-09-29 → 2023-09-28

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10470313

## Citation

> US National Institutes of Health, RePORTER application 10470313, Using human iPSC-derived microglia to understand the biological function of Alzheimer's risk variants (5F32AG069503-03). Retrieved via AI Analytics 2026-05-22 from https://api.ai-analytics.org/grant/nih/10470313. Licensed CC0.

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