# Induced neuronal cells: A novel tool to study neuropsychiatric diseases

> **NIH NIH R01** · STANFORD UNIVERSITY · 2022 · $736,190

## Abstract

Abstract
The goal of this project is to describe the function of synaptic adhesion molecules of the Neuroligin family
(Nlgns) in the mouse brain and in human neurons. Recent single cell expression studies have highlighted the
obversation that Nlgns are expressed also in non-neuronal cells, in particular oligodendrocyte precursors cells
(OPCs) and astrocytes who express Nlgns to even higher levels than neurons. Since little is known about the
function of Nlgns in glia and their effect on neurons and neural circuits, we propose to specifically delete Nlgns
in OPCs and astrocytes using our triple conditional Nlgn1-3 knock-out strain. Brains will be characterized
morphologically, electrophysiologically on the cellular and circuit level, and mutant mice will be characterized
by behavior. Next, we will perform an in-depth molecular characterization of the Neuroligin proteins by
characterizing the molecular mechanisms underlying the surprising functional diversity of Nlgns. We will map
their functional domains in mouse neurons by expressing various domain-mutant proteins in Nlgn1-4 quadruple
knock-out cells. We will explore whether Nlgn sequence relates to functional specificity and investigate the
notion of a synaptic Neurexin “code” that may determine Nlgn specificity.
 To complement our mouse studies and explore human-specific Neuroligin function as well as human
disease-associated mutations, we will capitalize on our previous human stem cell and reprogramming work in
which we have developed human induced neuronal (iN) cells that exhibit all principal functional properties of
primary mouse neurons including robust synapse formation. We propose to utilize this system to investigate
the so far obscure function of NLGN4Y, a Y chromosomal gene closely related to NLGN4 on the X-
chromosome and a member of the family not present in mouse. We will assess subcellular targeting by tagging
the endogenous locus and assess the functional consequences of genetic deletion. Another frequently mutated
Nlgn gene is NLGN3. Unlike NLGN4 it is better conserved in mice, but almost nothing is known about its
function in human cells. In addition to generate loss-of-function alleles, we will study the functional
consequences of distinct ASD-associated mutations introduced into the human NLGN3 gene. We will use a
conditional mutagenesis approach as we have successfully done in the past, as it allows the generation of a
perfect control conidition derived from the identical cell line as the experimental condition. Mutant human
neurons and controls will be characterized biochemically, morphologically, by gene expression, and
electrophysiologically. Finally, we propose to investigate the role of the proposed Nlgns-modulators MDGAs
which are also found mutated in ASD and other neurodevelopmental disorders. We will assess their
requirement for proper synapse formation and function by generating loss-of-function alleles in human
neurons. We will further probe their function as Neur...

## Key facts

- **NIH application ID:** 10469463
- **Project number:** 5R01MH092931-13
- **Recipient organization:** STANFORD UNIVERSITY
- **Principal Investigator:** Thomas C. Sudhof
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2022
- **Award amount:** $736,190
- **Award type:** 5
- **Project period:** 2010-09-29 → 2025-08-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10469463

## Citation

> US National Institutes of Health, RePORTER application 10469463, Induced neuronal cells: A novel tool to study neuropsychiatric diseases (5R01MH092931-13). Retrieved via AI Analytics 2026-05-22 from https://api.ai-analytics.org/grant/nih/10469463. Licensed CC0.

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