Genome‐wide profiling of N6‐methyladenosine‐modified pseudogene‐derived long noncoding RNAs reveals the tumour‐promoting and innate immune‐restraining function of RPS15AP12 in ovarian cancer

Abstract Background Pseudogene‐derived lncRNAs are widely dysregulated in cancer. Technological advancements have facilitated the functional characterization of increasing pseudogenes in cancer progression. However, the association between pseudogenes and RNA N6‐methyladenosine (m6A) modification in...

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Main Authors: Jie Xu, Yifei Ren, Jiayi Lu, Fengjiang Qin, Dan Yang, Chunyan Tang, Yu Yang, Jing Xu, Tao Liu, Ping Yi
Format: Article
Language:English
Published: Wiley 2025-03-01
Series:Clinical and Translational Medicine
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Online Access:https://doi.org/10.1002/ctm2.70249
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author Jie Xu
Yifei Ren
Jiayi Lu
Fengjiang Qin
Dan Yang
Chunyan Tang
Yu Yang
Jing Xu
Tao Liu
Ping Yi
author_facet Jie Xu
Yifei Ren
Jiayi Lu
Fengjiang Qin
Dan Yang
Chunyan Tang
Yu Yang
Jing Xu
Tao Liu
Ping Yi
author_sort Jie Xu
collection DOAJ
description Abstract Background Pseudogene‐derived lncRNAs are widely dysregulated in cancer. Technological advancements have facilitated the functional characterization of increasing pseudogenes in cancer progression. However, the association between pseudogenes and RNA N6‐methyladenosine (m6A) modification in cancer, as well as the underlying mechanisms, remains largely unexplored. Methods We analyzed the expression of 12 146 pseudogenes and comprehensively examined the m6A modification of RNAs derived from them and their paralogs. Through integrative analysis of multi‐omics data, we explored the associations between pseudogene dysregulation and m6A, identifying critical pseudogenes involved in HGSOC progression. Tumour promotion role of RPS15AP12 and its cognate parent gene was characterized by cell proliferation, transwell assays, and scratch assays in ovarian cells and xenograft nude mice. RNA decay assays were used to reveal the participation of m6A in decreasement of RPS15AP12 lncRNA stability. Luciferase reporter assays were performed to verify that RPS15AP12 enhances RPS15A expression by competitively binding to miR‐96‐3p. Western blot and phosphorylation assays were performed to investigate the impairment of RPS15AP12 towards the sensors of MAVS (RIG‐I and MDA5), and downstream p‐TBK1 and p‐IRF3. Finally, ELISA assays were performed to explore the regulatory role of RPS15AP12 in IFN‐β expression. Results M6A is distributed across over a thousand pseudogenes, and hypomethylation leads to their upregulation in HGSOC. We identified a processed pseudogene, RPS15AP12, upregulated by FTO‐mediated m6A demethylation. RPS15AP12 enhances the growth ability and metastatic capabilities of ovarian cancer (OC) cells via functioning as a competitive endogenous RNA (ceRNA) for its host gene, RPS15A, through the sequestration of miR‐96‐3p. Importantly, the deletion of RPS15AP12 diminishes the expression of RPS15A, leading to the upregulation of anti‐tumour immune responses by activating RIG‐I and MDA5 and downstream p‐TBK1 and p‐IRF3 as well as IFN‐β levels. Conclusion Our findings expand the understanding of m6A‐modulated pseudogenes in tumour growth and anti‐tumour innate immunity in OC. Key Points Genome‐wide profiling reveals the redistribution of m6A modification on pseudogene‐derived lncRNAs and m6A redistribution‐relevant dysregulation of pseudogenes in HGSOC. RPS15AP12, as a representative processed pseudogene, is up‐regulated by FTO‐mediated demethylation and acts as a miRNA sponge to promote RPS15A expression via competitively binding to miR‐96‐3p. RPS15AP12/RPS15A axis inhibits MAVS sensors (RIG‐I and MDA5) and downstream IFN‐β levels in ovarian cancer.
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spelling doaj-art-373799297eaa49c8b6bb0f1e2fe0e4ff2025-08-20T03:47:36ZengWileyClinical and Translational Medicine2001-13262025-03-01153n/an/a10.1002/ctm2.70249Genome‐wide profiling of N6‐methyladenosine‐modified pseudogene‐derived long noncoding RNAs reveals the tumour‐promoting and innate immune‐restraining function of RPS15AP12 in ovarian cancerJie Xu0Yifei Ren1Jiayi Lu2Fengjiang Qin3Dan Yang4Chunyan Tang5Yu Yang6Jing Xu7Tao Liu8Ping Yi9Department of Obstetrics and Gynecology The Third Affiliated Hospital of Chongqing Medical University Chongqing ChinaDepartment of Obstetrics and Gynecology The Third Affiliated Hospital of Chongqing Medical University Chongqing ChinaDepartment of Obstetrics and Gynecology The Third Affiliated Hospital of Chongqing Medical University Chongqing ChinaDepartment of Obstetrics and Gynecology The Third Affiliated Hospital of Chongqing Medical University Chongqing ChinaDepartment of Obstetrics and Gynecology The Third Affiliated Hospital of Chongqing Medical University Chongqing ChinaDepartment of Obstetrics and Gynecology The Third Affiliated Hospital of Chongqing Medical University Chongqing ChinaDepartment of Obstetrics and Gynecology The Third Affiliated Hospital of Chongqing Medical University Chongqing ChinaDepartment of Obstetrics and Gynecology The Third Affiliated Hospital of Chongqing Medical University Chongqing ChinaDepartment of Obstetrics and Gynecology The Third Affiliated Hospital of Chongqing Medical University Chongqing ChinaDepartment of Obstetrics and Gynecology The Third Affiliated Hospital of Chongqing Medical University Chongqing ChinaAbstract Background Pseudogene‐derived lncRNAs are widely dysregulated in cancer. Technological advancements have facilitated the functional characterization of increasing pseudogenes in cancer progression. However, the association between pseudogenes and RNA N6‐methyladenosine (m6A) modification in cancer, as well as the underlying mechanisms, remains largely unexplored. Methods We analyzed the expression of 12 146 pseudogenes and comprehensively examined the m6A modification of RNAs derived from them and their paralogs. Through integrative analysis of multi‐omics data, we explored the associations between pseudogene dysregulation and m6A, identifying critical pseudogenes involved in HGSOC progression. Tumour promotion role of RPS15AP12 and its cognate parent gene was characterized by cell proliferation, transwell assays, and scratch assays in ovarian cells and xenograft nude mice. RNA decay assays were used to reveal the participation of m6A in decreasement of RPS15AP12 lncRNA stability. Luciferase reporter assays were performed to verify that RPS15AP12 enhances RPS15A expression by competitively binding to miR‐96‐3p. Western blot and phosphorylation assays were performed to investigate the impairment of RPS15AP12 towards the sensors of MAVS (RIG‐I and MDA5), and downstream p‐TBK1 and p‐IRF3. Finally, ELISA assays were performed to explore the regulatory role of RPS15AP12 in IFN‐β expression. Results M6A is distributed across over a thousand pseudogenes, and hypomethylation leads to their upregulation in HGSOC. We identified a processed pseudogene, RPS15AP12, upregulated by FTO‐mediated m6A demethylation. RPS15AP12 enhances the growth ability and metastatic capabilities of ovarian cancer (OC) cells via functioning as a competitive endogenous RNA (ceRNA) for its host gene, RPS15A, through the sequestration of miR‐96‐3p. Importantly, the deletion of RPS15AP12 diminishes the expression of RPS15A, leading to the upregulation of anti‐tumour immune responses by activating RIG‐I and MDA5 and downstream p‐TBK1 and p‐IRF3 as well as IFN‐β levels. Conclusion Our findings expand the understanding of m6A‐modulated pseudogenes in tumour growth and anti‐tumour innate immunity in OC. Key Points Genome‐wide profiling reveals the redistribution of m6A modification on pseudogene‐derived lncRNAs and m6A redistribution‐relevant dysregulation of pseudogenes in HGSOC. RPS15AP12, as a representative processed pseudogene, is up‐regulated by FTO‐mediated demethylation and acts as a miRNA sponge to promote RPS15A expression via competitively binding to miR‐96‐3p. RPS15AP12/RPS15A axis inhibits MAVS sensors (RIG‐I and MDA5) and downstream IFN‐β levels in ovarian cancer.https://doi.org/10.1002/ctm2.70249innate immune responsem6AmiRNA spongeovarian cancerpseudogene
spellingShingle Jie Xu
Yifei Ren
Jiayi Lu
Fengjiang Qin
Dan Yang
Chunyan Tang
Yu Yang
Jing Xu
Tao Liu
Ping Yi
Genome‐wide profiling of N6‐methyladenosine‐modified pseudogene‐derived long noncoding RNAs reveals the tumour‐promoting and innate immune‐restraining function of RPS15AP12 in ovarian cancer
Clinical and Translational Medicine
innate immune response
m6A
miRNA sponge
ovarian cancer
pseudogene
title Genome‐wide profiling of N6‐methyladenosine‐modified pseudogene‐derived long noncoding RNAs reveals the tumour‐promoting and innate immune‐restraining function of RPS15AP12 in ovarian cancer
title_full Genome‐wide profiling of N6‐methyladenosine‐modified pseudogene‐derived long noncoding RNAs reveals the tumour‐promoting and innate immune‐restraining function of RPS15AP12 in ovarian cancer
title_fullStr Genome‐wide profiling of N6‐methyladenosine‐modified pseudogene‐derived long noncoding RNAs reveals the tumour‐promoting and innate immune‐restraining function of RPS15AP12 in ovarian cancer
title_full_unstemmed Genome‐wide profiling of N6‐methyladenosine‐modified pseudogene‐derived long noncoding RNAs reveals the tumour‐promoting and innate immune‐restraining function of RPS15AP12 in ovarian cancer
title_short Genome‐wide profiling of N6‐methyladenosine‐modified pseudogene‐derived long noncoding RNAs reveals the tumour‐promoting and innate immune‐restraining function of RPS15AP12 in ovarian cancer
title_sort genome wide profiling of n6 methyladenosine modified pseudogene derived long noncoding rnas reveals the tumour promoting and innate immune restraining function of rps15ap12 in ovarian cancer
topic innate immune response
m6A
miRNA sponge
ovarian cancer
pseudogene
url https://doi.org/10.1002/ctm2.70249
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