# Dissecting the role of NOTCH2NL genes in human brain development and neurological disorders associated with chromosome 1q21.1 distal duplications and deletions.

> **NIH NIH R01** · UNIVERSITY OF CALIFORNIA SANTA CRUZ · 2022 · $726,976

## Abstract

Genomic copy number variation (CNV) in a part of human chromosome 1 emerged recently in the
human lineage by repeated segmental duplications and rearrangements. CNVs here have been associated
with autism, schizophrenia and other neurodevelopmental disorders in multiple genome-wide association
studies. However, the gene(s) in this interval that contribute to these phenotypes have not been established.
We recently identified a family of 3 genes, ​NOTCH2NLA​, -​B and -​C that reside in this locus, are highly
expressed during early brain development and are capable of promoting cortical neuron stem cell maintenance
and proliferation. In this proposal ​we test the hypothesis that NOTCH2NL genes are required for normal
human brain development and alterations in their gene dosage contribute the neurological phenotypes
observed in patients with 1q21.1 distal deletions and duplications. This will be accomplished in Aim 1 by ​resequencing this genomic interval ​in diverse human population samples using new long genomic fragment sequencing technologies that will enable us ​to identify the NOTCH2NL alleles present in the human population ​and structural variation present in this highly repetitive genomic region​. This information will inform ​subsequent sequence analysis of 14 samples
harboring pathogenic 1q21.1 CNV events that may implicate specific NOTCH2NL loci and variants in
these disorders. This information will be used to develop ​high throughput, cost-effective assays to
identify the specific NOTCH2NL alleles present in large genomic DNA collections from patients with
1q21.1-associated neurological disorders. In Aim 2, we will ​test the activity of the various NOTCH2NL
alleles identified in 2 ways. First, examine the ability of each NOTCH2NL allele ​to promote NOTCH signaling
using reporter based assays​. Second, we will test the activity of specific NOTCH2NL alleles ​to promote
normal cortical organoid formation using isogenic CRISPR engineered pluripotent stem cell lines that
differ only by the specific NOTCH2NL alleles present. The ability of specific alleles to rescue defects in the
balance of neural stem cells and cortical neurons associated with loss of NOTCH2NL will be measured by
single-cell RNA Sequencing, bulk gene expression measurements and histology. Finally, in Aim 3 we will test
whether ​heterozygous, large-scale 1q21.1 deletions similar to those observed in patients have similar
or more severe defects in early brain development as assayed by our human pluripotent stem cell
cerebral cortex organoid assay using the analysis methods described for Aim 2. We will then ​test the ability
of NOTCH2NL and other genes in locus to rescue any defects observed. The experiments outlined here will improve our understanding of the specific biological defects resulting from 1q21.1 CNVs that ultimately lead to complex neurological disorders. These methods can be applied to other repetitive genomic loci implicated in neurodevelopmental diseases.

## Key facts

- **NIH application ID:** 10333371
- **Project number:** 5R01MH120295-03
- **Recipient organization:** UNIVERSITY OF CALIFORNIA SANTA CRUZ
- **Principal Investigator:** Sofie Reda Salama
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2022
- **Award amount:** $726,976
- **Award type:** 5
- **Project period:** 2020-04-15 → 2025-01-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10333371

## Citation

> US National Institutes of Health, RePORTER application 10333371, Dissecting the role of NOTCH2NL genes in human brain development and neurological disorders associated with chromosome 1q21.1 distal duplications and deletions. (5R01MH120295-03). Retrieved via AI Analytics 2026-05-22 from https://api.ai-analytics.org/grant/nih/10333371. Licensed CC0.

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