# The role of N6-methyladenosine RNA modification in programmed and aberrant DNA mutagenesis in B cells

> **NIH NIH R01** · COLUMBIA UNIVERSITY HEALTH SCIENCES · 2022 · $509,471

## Abstract

PROJECT SUMMARY
Background: VDJ recombination, Class switch recombination (CSR) and somatic hypermutation (SHM) are
three B lymphocyte specific processes that mediate antibody gene diversification. VDJ recombination requires
the DNA double strand generation by the Recombination activation genes (RAG1 an RAG2) where as CSR and
SHM requires the single-strand DNA break activity of the Activation Induced Deaminase (AID) enzyme. Both
RAG1/2 and AID activities are coupled with noncoding RNA transcription at sites of DNA break/mutation. The
properties of the ncRNAs generated at sites of programmed DNA breaks are poorly characterized in B cells.
Recently advances in biology has provided compelling evidence that post-transcriptional and co-transcriptional
modification of ncRNAs determine a component of RNA epigenomics and have significant role in driving
cellular development and function. In this application, supported by preliminary data generated in our
laboratory, we are evaluating the role of RNA modification N6-methyladenosine (m6A) and its associated
enzymes METTL3 and METTL14 in B cell development, function and genomic integrity.
Objective/Hypothesis: In his proposal, we will determine how RNA methylation m6A on transcripts
generated in the IgH locus and the rest of the B cell genome controls programmed DNA recombination,
antibody gene diversification and prevents chromosomal instability.
Specific Aims: Aim 1: Aim 1: Are m6A modifying enzymes Mettl3 and Mettl14 important for class switch
recombination, somatic hypermutation, and/or for preventing genomic stability in B cells? ; AIM 2: What is the
mechanism by which m6A modification promotes IgH DNA recombination and/or prevents genomic
instability? AIM 3: Do RNA methylation and coupled RNA surveillance pathways have a role in early B cell
development and during VDJ recombination?
Study Design: Using cell lines and mouse models, we will study mechanism by which RNA m6A methylation
plays a role in programmed DNA recombination and protection of B cell genomic integrity. We will evaluate
the mechanism of formation of single-strand DNA structures formation and RNA surveillance at sites of AID
activity in matured B cells and the mechanism by which inhibitory RNAs are degraded at sites of RAG activity
in B cells during VDJ recombination. We will also evaluate the molecular mechanism by which RNA
modification promotes programmed DNA rearrangements. We use a combination of mouse genetics, genomics,
biochemistry and 3D-STORM microscopy to accomplish the goals of the proposed project.
Disease Relevance: B cells are a central component of the adaptive immune response, but also prone to
undergo leukemias and lymphomas when antibody gene diversification processes are not well controlled.
Proposed studies leads to a better understanding of the mechanism of antibody gene diversification but also
educate us how B cell cancer (specially in context of DLBCL and Multiple Myeloma) are prevented.

## Key facts

- **NIH application ID:** 10461710
- **Project number:** 5R01AI143897-03
- **Recipient organization:** COLUMBIA UNIVERSITY HEALTH SCIENCES
- **Principal Investigator:** Uttiya Basu
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2022
- **Award amount:** $509,471
- **Award type:** 5
- **Project period:** 2020-09-09 → 2024-08-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10461710

## Citation

> US National Institutes of Health, RePORTER application 10461710, The role of N6-methyladenosine RNA modification in programmed and aberrant DNA mutagenesis in B cells (5R01AI143897-03). Retrieved via AI Analytics 2026-05-24 from https://api.ai-analytics.org/grant/nih/10461710. Licensed CC0.

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