Depletion of METTL3 alters cellular and extracellular levels of miRNAs containing m6A consensus sequences
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 dis...
| Main Authors: | , , , , , , , , , , , , , , |
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| Other Authors: | |
| Format: | Article |
| Language: | English |
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Elsevier BV
2023
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| Online Access: | https://hdl.handle.net/1721.1/147799 |
| _version_ | 1811081434325057536 |
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| author | 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 Vickers, Kasey C Dedon, Peter Weaver, Alissa M Coffey, Robert J Patton, James G |
| author2 | Massachusetts Institute of Technology. Department of Biological Engineering |
| author_facet | Massachusetts Institute of Technology. Department of Biological Engineering 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 Vickers, Kasey C Dedon, Peter Weaver, Alissa M Coffey, Robert J Patton, James G |
| author_sort | Abner, Jessica J |
| collection | MIT |
| description | 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. |
| first_indexed | 2024-09-23T11:46:37Z |
| format | Article |
| id | mit-1721.1/147799 |
| institution | Massachusetts Institute of Technology |
| language | English |
| last_indexed | 2024-09-23T11:46:37Z |
| publishDate | 2023 |
| publisher | Elsevier BV |
| record_format | dspace |
| spelling | mit-1721.1/1477992023-02-01T03:34:13Z Depletion of METTL3 alters cellular and extracellular levels of miRNAs containing m6A consensus sequences 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 Vickers, Kasey C Dedon, Peter Weaver, Alissa M Coffey, Robert J Patton, James G Massachusetts Institute of Technology. Department of Biological Engineering 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. 2023-01-31T15:01:32Z 2023-01-31T15:01:32Z 2021 2023-01-31T14:57:26Z Article http://purl.org/eprint/type/JournalArticle https://hdl.handle.net/1721.1/147799 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). en 10.1016/J.HELIYON.2021.E08519 Heliyon Creative Commons Attribution-NonCommercial-NoDerivs License http://creativecommons.org/licenses/by-nc-nd/4.0/ application/pdf Elsevier BV Elsevier |
| spellingShingle | 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 Vickers, Kasey C Dedon, Peter Weaver, Alissa M Coffey, Robert J Patton, James G Depletion of METTL3 alters cellular and extracellular levels of miRNAs containing m6A consensus sequences |
| title | Depletion of METTL3 alters cellular and extracellular levels of miRNAs containing m6A consensus sequences |
| title_full | Depletion of METTL3 alters cellular and extracellular levels of miRNAs containing m6A consensus sequences |
| title_fullStr | Depletion of METTL3 alters cellular and extracellular levels of miRNAs containing m6A consensus sequences |
| title_full_unstemmed | Depletion of METTL3 alters cellular and extracellular levels of miRNAs containing m6A consensus sequences |
| title_short | Depletion of METTL3 alters cellular and extracellular levels of miRNAs containing m6A consensus sequences |
| title_sort | depletion of mettl3 alters cellular and extracellular levels of mirnas containing m6a consensus sequences |
| url | https://hdl.handle.net/1721.1/147799 |
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