Genetic and pharmacological inhibition of METTL3 alleviates renal fibrosis by reducing EVL m6A modification through an IGF2BP2‐dependent mechanism
Abstract Background N6‐methyladenosine (m6A) is of great importance in renal physiology and disease progression, but its function and mechanism in renal fibrosis remain to be comprehensively and extensively explored. Hence, this study will explore the function and potential mechanism of critical reg...
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Wiley
2023-08-01
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| Series: | Clinical and Translational Medicine |
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| Online Access: | https://doi.org/10.1002/ctm2.1359 |
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| author | Wei‐Jian Ni Hong Zhou Hao Lu Nan‐Nan Ma Bing‐Bing Hou Wei Li Fan‐Xu Kong Ju‐Tao Yu Rui Hou Juan Jin Jia‐Gen Wen Tao Zhang Xiao‐Ming Meng |
| author_facet | Wei‐Jian Ni Hong Zhou Hao Lu Nan‐Nan Ma Bing‐Bing Hou Wei Li Fan‐Xu Kong Ju‐Tao Yu Rui Hou Juan Jin Jia‐Gen Wen Tao Zhang Xiao‐Ming Meng |
| author_sort | Wei‐Jian Ni |
| collection | DOAJ |
| description | Abstract Background N6‐methyladenosine (m6A) is of great importance in renal physiology and disease progression, but its function and mechanism in renal fibrosis remain to be comprehensively and extensively explored. Hence, this study will explore the function and potential mechanism of critical regulator‐mediated m6A modification during renal fibrosis and thereby explore promising anti‐renal fibrosis agents. Methods Renal tissues from humans and mice as well as HK‐2 cells were used as research subjects. The profiles of m6A modification and regulators in renal fibrosis were analysed at the protein and RNA levels using Western blotting, quantitative real‐time polymerase chain reaction and other methods. Methylation RNA immunoprecipitation sequencing and RNA sequencing coupled with methyltransferase‐like 3 (METTL3) conditional knockout were used to explore the function of METTL3 and potential targets. Gene silencing and overexpression combined with RNA immunoprecipitation were performed to investigate the underlying mechanism by which METTL3 regulates the Ena/VASP‐like (EVL) m6A modification that promotes renal fibrosis. Molecular docking and virtual screening with in vitro and in vivo experiments were applied to screen promising traditional Chinese medicine (TCM) monomers and explore their mechanism of regulating the METTL3/EVL m6A axis and anti‐renal fibrosis. Results METTL3 and m6A modifications were hyperactivated in both the tubular region of fibrotic kidneys and HK‐2 cells. Upregulated METTL3 enhanced the m6A modification of EVL mRNA to improve its stability and expression in an insulin‐like growth factor 2 mRNA‐binding protein 2 (IGF2BP2)‐dependent manner. Highly expressed EVL binding to Smad7 abrogated the Smad7‐induced suppression of transforming growth factor‐β (TGF‐β1)/Smad3 signal transduction, which conversely facilitated renal fibrosis progression. Molecular docking and virtual screening based on the structure of METTL3 identified a TCM monomer named isoforsythiaside, which inhibited METTL3 activity together with the METTL3/EVL m6A axis to exert anti‐renal fibrosis effects. Conclusions Collectively, the overactivated METTL3/EVL m6A axis is a potential target for renal fibrosis therapy, and the pharmacological inhibition of METTL3 activity by isoforsythiaside suggests that it is a promising anti‐renal fibrosis agent. |
| first_indexed | 2024-03-11T22:49:27Z |
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| spelling | doaj.art-20fbaf649a3b4d6f87fb805cc65e76ca2023-09-22T04:05:54ZengWileyClinical and Translational Medicine2001-13262023-08-01138n/an/a10.1002/ctm2.1359Genetic and pharmacological inhibition of METTL3 alleviates renal fibrosis by reducing EVL m6A modification through an IGF2BP2‐dependent mechanismWei‐Jian Ni0Hong Zhou1Hao Lu2Nan‐Nan Ma3Bing‐Bing Hou4Wei Li5Fan‐Xu Kong6Ju‐Tao Yu7Rui Hou8Juan Jin9Jia‐Gen Wen10Tao Zhang11Xiao‐Ming Meng12Department of Pharmacy Anhui Provincial Hospital, The First Affiliated Hospital of USTC, Division of Life Sciences and Medicine University of Science and Technology of China HefeiAnhuiPeople's Republic of ChinaDepartment of Pharmacy Anhui Provincial Cancer Hospital, The First Affiliated Hospital of USTC, Division of Life Sciences and Medicine University of Science and Technology of China HefeiAnhuiPeople's Republic of ChinaInflammation and Immune Mediated Diseases Laboratory of Anhui Province Anhui Institute of Innovative Drugs School of Pharmacy Anhui Medical University HefeiAnhuiPeople's Republic of ChinaDepartment of Urology The Second Affiliated Hospital of Anhui Medical University HefeiAnhuiPeople's Republic of ChinaDepartment of Urology The First Affiliated Hospital of Anhui Medical University HefeiAnhuiPeople's Republic of ChinaInflammation and Immune Mediated Diseases Laboratory of Anhui Province Anhui Institute of Innovative Drugs School of Pharmacy Anhui Medical University HefeiAnhuiPeople's Republic of ChinaInflammation and Immune Mediated Diseases Laboratory of Anhui Province Anhui Institute of Innovative Drugs School of Pharmacy Anhui Medical University HefeiAnhuiPeople's Republic of ChinaInflammation and Immune Mediated Diseases Laboratory of Anhui Province Anhui Institute of Innovative Drugs School of Pharmacy Anhui Medical University HefeiAnhuiPeople's Republic of ChinaInflammation and Immune Mediated Diseases Laboratory of Anhui Province Anhui Institute of Innovative Drugs School of Pharmacy Anhui Medical University HefeiAnhuiPeople's Republic of ChinaResearch Center for Translational Medicine The Second Affiliated Hospital of Anhui Medical University HefeiAnhuiPeople's Republic of ChinaInflammation and Immune Mediated Diseases Laboratory of Anhui Province Anhui Institute of Innovative Drugs School of Pharmacy Anhui Medical University HefeiAnhuiPeople's Republic of ChinaDepartment of Urology The Second Affiliated Hospital of Anhui Medical University HefeiAnhuiPeople's Republic of ChinaInflammation and Immune Mediated Diseases Laboratory of Anhui Province Anhui Institute of Innovative Drugs School of Pharmacy Anhui Medical University HefeiAnhuiPeople's Republic of ChinaAbstract Background N6‐methyladenosine (m6A) is of great importance in renal physiology and disease progression, but its function and mechanism in renal fibrosis remain to be comprehensively and extensively explored. Hence, this study will explore the function and potential mechanism of critical regulator‐mediated m6A modification during renal fibrosis and thereby explore promising anti‐renal fibrosis agents. Methods Renal tissues from humans and mice as well as HK‐2 cells were used as research subjects. The profiles of m6A modification and regulators in renal fibrosis were analysed at the protein and RNA levels using Western blotting, quantitative real‐time polymerase chain reaction and other methods. Methylation RNA immunoprecipitation sequencing and RNA sequencing coupled with methyltransferase‐like 3 (METTL3) conditional knockout were used to explore the function of METTL3 and potential targets. Gene silencing and overexpression combined with RNA immunoprecipitation were performed to investigate the underlying mechanism by which METTL3 regulates the Ena/VASP‐like (EVL) m6A modification that promotes renal fibrosis. Molecular docking and virtual screening with in vitro and in vivo experiments were applied to screen promising traditional Chinese medicine (TCM) monomers and explore their mechanism of regulating the METTL3/EVL m6A axis and anti‐renal fibrosis. Results METTL3 and m6A modifications were hyperactivated in both the tubular region of fibrotic kidneys and HK‐2 cells. Upregulated METTL3 enhanced the m6A modification of EVL mRNA to improve its stability and expression in an insulin‐like growth factor 2 mRNA‐binding protein 2 (IGF2BP2)‐dependent manner. Highly expressed EVL binding to Smad7 abrogated the Smad7‐induced suppression of transforming growth factor‐β (TGF‐β1)/Smad3 signal transduction, which conversely facilitated renal fibrosis progression. Molecular docking and virtual screening based on the structure of METTL3 identified a TCM monomer named isoforsythiaside, which inhibited METTL3 activity together with the METTL3/EVL m6A axis to exert anti‐renal fibrosis effects. Conclusions Collectively, the overactivated METTL3/EVL m6A axis is a potential target for renal fibrosis therapy, and the pharmacological inhibition of METTL3 activity by isoforsythiaside suggests that it is a promising anti‐renal fibrosis agent.https://doi.org/10.1002/ctm2.1359chronic kidney diseaseinsulin‐like growth factor 2 mRNA‐binding protein 2 (IGF2BP2)methyltransferase‐like 3 (METTL3)N6‐methyladenosine (m6A)renal fibrosistraditional Chinese medicine |
| spellingShingle | Wei‐Jian Ni Hong Zhou Hao Lu Nan‐Nan Ma Bing‐Bing Hou Wei Li Fan‐Xu Kong Ju‐Tao Yu Rui Hou Juan Jin Jia‐Gen Wen Tao Zhang Xiao‐Ming Meng Genetic and pharmacological inhibition of METTL3 alleviates renal fibrosis by reducing EVL m6A modification through an IGF2BP2‐dependent mechanism Clinical and Translational Medicine chronic kidney disease insulin‐like growth factor 2 mRNA‐binding protein 2 (IGF2BP2) methyltransferase‐like 3 (METTL3) N6‐methyladenosine (m6A) renal fibrosis traditional Chinese medicine |
| title | Genetic and pharmacological inhibition of METTL3 alleviates renal fibrosis by reducing EVL m6A modification through an IGF2BP2‐dependent mechanism |
| title_full | Genetic and pharmacological inhibition of METTL3 alleviates renal fibrosis by reducing EVL m6A modification through an IGF2BP2‐dependent mechanism |
| title_fullStr | Genetic and pharmacological inhibition of METTL3 alleviates renal fibrosis by reducing EVL m6A modification through an IGF2BP2‐dependent mechanism |
| title_full_unstemmed | Genetic and pharmacological inhibition of METTL3 alleviates renal fibrosis by reducing EVL m6A modification through an IGF2BP2‐dependent mechanism |
| title_short | Genetic and pharmacological inhibition of METTL3 alleviates renal fibrosis by reducing EVL m6A modification through an IGF2BP2‐dependent mechanism |
| title_sort | genetic and pharmacological inhibition of mettl3 alleviates renal fibrosis by reducing evl m6a modification through an igf2bp2 dependent mechanism |
| topic | chronic kidney disease insulin‐like growth factor 2 mRNA‐binding protein 2 (IGF2BP2) methyltransferase‐like 3 (METTL3) N6‐methyladenosine (m6A) renal fibrosis traditional Chinese medicine |
| url | https://doi.org/10.1002/ctm2.1359 |
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