# The in vivo role of m6A RNA modification in Cellular Stress, Aging, and Neurological ­Disease

> **NIH NIH F31** · UNIVERSITY OF PENNSYLVANIA · 2020 · $45,520

## Abstract

Project Summary
RNA modifications embody a crucial layer of epigenetic regulation of gene expression1,2. N6-methyladenosine
RNA (m6A) is the most prevalent modification present on eukaryotic mRNA, and has been shown to modulate
several post-transcriptional processes such as mRNA splicing, mRNA decay, secondary structure, and
translation1,2. Recent studies have uncovered critical roles of m6A modification in developmental processes, cell
differentiation, cancer metastasis, and cellular stress responses3-13. Mechanistic studies in mammalian cell lines
suggest that m6A deposition is critical during the cellular stress response, where methylation has been shown to
control selective translation and stress granule localization8-13,46. However, the role of m6A in cellular stress in
vivo and in the aged brain has not been addressed. Given the crucial role of the stress response in brain aging
and neurodegenerative disease, m6A modification of RNA may play an important role. To elucidate this role, in
Aim 1 I propose to address the functional consequence of increased m6A mRNA modification (seen by dot blot
analysis) in the chronic stress of aging in Drosophila brains. This would be the first in vivo analysis of m6A
dynamics in the brain during aging. I will assess which specific transcripts show changed levels of m6A
modification by performing m6A-miCLIP sequencing from brains of 3-d young and 30-d aged flies. To further
understand the consequence of altered modification levels on the transcript I will also examine transcript levels
by RNA-seq, and translation efficiency by ribosomal sequencing in control and m6A methyltransferase (Ime4)
knockdown fly brains in young and aged flies. Furthermore, my preliminary data suggest that Ime4 knockdown
flies cause a decrease in lifespan. I will examine also if Ime4 RNAi flies exhibit increased vacuole degeneration
from paraffin sectioning of brains. Aim 2 will examine the role of m6A RNA reader protein CG6422 in
neurodegeneration, specifically TDP-43 induced degeneration. Preliminary data shows that TDP-43 expressing
fly heads have increased levels of m6A methylated mRNA than controls. Furthermore, ubiquitous reduction of
the m6A cytoplasmic reader protein CG6422 in flies which express TDP-43 dramatically reduces lifespan in
comparison to TDP-43 expressing flies. It is likely that CG6422 is playing a role in the pathological stress
response caused by TDP-43 toxicity, and upregulation of this gene may have therapeutic benefits. To further
understand this role, I will examine if the decrease in lifespan also exists with a neuronal specific expression of
TDP-43 and CG6422 RNAi, examine retinal degeneration with CG6422 RNAi, and possible interaction of
CG6422 with TDP-43. I will further examine the effect CG6422 may be playing on stress regulation processes
induced during neurodegeneration such as heat shock protein levels and stress granule dynamics. By examining
m6A regulation in aged flies or fly models of degenerat...

## Key facts

- **NIH application ID:** 10020161
- **Project number:** 5F31AG063470-02
- **Recipient organization:** UNIVERSITY OF PENNSYLVANIA
- **Principal Investigator:** Alexandra Emily Perlegos
- **Activity code:** F31 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2020
- **Award amount:** $45,520
- **Award type:** 5
- **Project period:** 2019-08-01 → 2022-07-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10020161

## Citation

> US National Institutes of Health, RePORTER application 10020161, The in vivo role of m6A RNA modification in Cellular Stress, Aging, and Neurological ­Disease (5F31AG063470-02). Retrieved via AI Analytics 2026-05-22 from https://api.ai-analytics.org/grant/nih/10020161. Licensed CC0.

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