# Quantitative mass spectrometry for comprehending epigenetic mechanisms in a new underlying neurological developmental disorder

> **NIH NIH R01** · WASHINGTON UNIVERSITY · 2024 · $512,247

## Abstract

Project Summary
Epigenetic mechanisms have been linked to many human disorders and diseases, most notably cancer.
However, in recent years more and more human diseases have been found with possible epigenetic insults. In
this regard, our collaborators at the Children’s Hospital of Philadelphia have through exome sequencing
discovered germline-line mutations to the H3FA and H3FB genes in pediatric patients that suffer from similar
neurological disorders and craniofacial abnormalities. These two genes encode for the histone variant H3.3, and
thus represent the first germ-line mutations found on histone H3 in humans (neurohistone H3.3 mutations).
Histones are small basic proteins that bind DNA to give rise to our chromatin structure. Along with DNA
methylation and long non-coding RNA, post-translational modifications to histones regulate gene expression
patterns (epigenetic mechanisms) and chromatin organization. Histone H3.3 is a specialized histone variant
linked to active genes, and somatic mutations to this histone have been found in different brain cancers
exclusively on the N-terminal tail. Our preliminary data have found that these neurohistone H3.3 mutations are
spread across the entire protein from the N- to C-terminus. Therefore, we hypothesize that these mutations while
resulting in similar phenotypes, do this by disrupting different epigenetic mechanisms involving histone H3.3. In
this proposal, we aim to understand how these neurohistone mutations lead to neurodevelopmental problems.
First, we aim to determine if these neurohistone mutations affect global or local histone modification patterns on
wild-type or mutant histone variants using novel mass spectrometry (MS) approaches in cellular models. Next,
we look to determine if these neurohistone mutants affect chromatin structure or conformation, or cause
misincorporation of the H3.3 variant in the genome. Lastly, we will characterize proteome expression in a mouse
model of one of the mutations using a novel in utero stable isotope labeling approach. It is our goal to determine
how these neurohistone H3.3 mutations affect epigenetic and cellular signaling mechanisms to disrupt
neurodevelopment and maintenance leading to neurological disorders in these pediatric patients.

## Key facts

- **NIH application ID:** 10890737
- **Project number:** 5R01HD106051-03
- **Recipient organization:** WASHINGTON UNIVERSITY
- **Principal Investigator:** Benjamin A Garcia
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2024
- **Award amount:** $512,247
- **Award type:** 5
- **Project period:** 2022-08-16 → 2027-07-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10890737

## Citation

> US National Institutes of Health, RePORTER application 10890737, Quantitative mass spectrometry for comprehending epigenetic mechanisms in a new underlying neurological developmental disorder (5R01HD106051-03). Retrieved via AI Analytics 2026-05-25 from https://api.ai-analytics.org/grant/nih/10890737. Licensed CC0.

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