Depletion of METTL3 alters cellular and extracellular levels of miRNAs containing m6A consensus sequences
Name
1-s2.0-S2405844021026220-main.pdf
Description
Published version
Size
1.37 MB
Format
Adobe PDF
Checksum (MD5)
b8ba3c6df71f210d84338ff91864d72f
Author(s)
Abner, Jessica J
Franklin, Jeffrey L
Clement, Margaret A
Hinger, Scott A
Allen, Ryan M
Liu, Xiao
Kellner, Stefanie
Wu, Junzhou
Karijolich, John
Liu, Qi
Date Issued
2021
Journal
Heliyon
Publisher
Elsevier BV
Citation
Abner, Jessica J, Franklin, Jeffrey L, Clement, Margaret A, Hinger, Scott A, Allen, Ryan M et al. 2021. "Depletion of METTL3 alters cellular and extracellular levels of miRNAs containing m6A consensus sequences." Heliyon, 7 (12).
Version
Final published version
Abstract
Extracellular vesicles (EVs) are capable of transferring cargo from donor to recipient cells, but precisely how cargo content is regulated for export is mostly unknown. For miRNA cargo, we previously showed that when compared to isogenic colorectal cancer (CRC) cells expressing wild-type KRAS, a distinct subset of miRNAs are differentially enriched in EVs from KRAS mutant active CRC cells, with miR-100 being one of the most enriched. The mechanisms that could explain how miR-100 and other miRNAs are differentially exported into EVs have not been fully elucidated. Here, we tested the effect of N6-methyladenosine (m6A) modification on miRNA export into EVs by depletion of METTL3 and ALKBH5, a writer and eraser of m6A modification, respectively. While the effects of ALKBH5 knockdown were quite modest, decreased levels of METTL3 led to reduced cellular and extracellular levels of a subset of miRNAs that contain consensus sequences for m6A modification. Functional testing of EVs prepared from cells expressing shRNAs against METTL3 showed that they were less capable of conferring colony growth in 3D to wild-type KRAS cells and were also largely incapable of conferring the spread of cetuximab resistance. Our data support a role for METTL3 modification on cellular miRNA levels and export of specific miRNAs.
MIT Department
Massachusetts Institute of Technology. Department of Biological Engineering
Terms of Use
Creative Commons Attribution-NonCommercial-NoDerivs License
Persistent DSpace Link
DOI of Published Version
https://doi.org/10.1016/J.HELIYON.2021.E08519
