Targeting the m6A RNA methyltransferase METTL3 attenuates the development of kidney fibrosis
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Kidney fibrosis is a major mechanism underlying chronic kidney disease (CKD). N6-methyladenosine (m6A) RNA methylation is associated with organ fibrosis. We investigated m6A profile alterations and the inhibitory effect of RNA methylation in kidney fibrosis in vitro (TGF-β-treated HK-2 cells) and in vivo (unilateral ureteral obstruction [UUO] mouse model). METTL3-mediated signaling was inhibited using siRNA in vitro or the METTL3-specific inhibitor STM2457 in vivo and in vitro. In HK-2 cells, METTL3 protein levels increased in a dose- and time-dependent manner along with an increase in the cellular m6A levels. In the UUO model, METTL3 expression and m6A levels were significantly increased. Transcriptomic and m6A profiling demonstrated that epithelial-to-mesenchymal transition- and inflammation-related pathways were significantly associated with RNA m6A methylation. Genetic and pharmacologic inhibition of METTL3 in HK-2 cells decreased TGF-β-induced fibrotic marker expression. STM2457-induced inhibition of METTL3 attenuated the degree of kidney fibrosis in vivo. Furthermore, METTL3 protein expression was significantly increased in the tissues of CKD patients with diabetic or IgA nephropathy. Therefore, targeting alterations in RNA methylation could be a potential therapeutic strategy for treating kidney fibrosis.
Abstract
Kidney fibrosis is a major mechanism underlying chronic kidney disease (CKD). N6-methyladenosine (m6A) RNA methylation is associated with organ fibrosis. We investigated m6A profile alterations and the inhibitory effect of RNA methylation in kidney fibrosis in vitro (TGF-β-treated HK-2 cells) and in vivo (unilateral ureteral obstruction [UUO] mouse model). METTL3-mediated signaling was inhibited using siRNA in vitro or the METTL3-specific inhibitor STM2457 in vivo and in vitro. In HK-2 cells, METTL3 protein levels increased in a dose- and time-dependent manner along with an increase in the cellular m6A levels. In the UUO model, METTL3 expression and m6A levels were significantly increased. Transcriptomic and m6A profiling demonstrated that epithelial-to-mesenchymal transition- and inflammation-related pathways were significantly associated with RNA m6A methylation. Genetic and pharmacologic inhibition of METTL3 in HK-2 cells decreased TGF-β-induced fibrotic marker expression. STM2457-induced inhibition of METTL3 attenuated the degree of kidney fibrosis in vivo. Furthermore, METTL3 protein expression was significantly increased in the tissues of CKD patients with diabetic or IgA nephropathy. Therefore, targeting alterations in RNA methylation could be a potential therapeutic strategy for treating kidney fibrosis.
참고문헌 (42)
- GBD, "Global, regional, and national burden of chronic kidney disease, 1990-2017:a systematic analysis for the Global Burden of Disease Study 2017", Lancet, 395, pp.709~733 (2020)
- Hill, N. R., "Global prevalence of chronic kidney disease—a systematic review and meta-analysis", PLoS ONE, 11, pp.e0158765~ (2016)
- Chen, T. K., "Chronic kidney disease diagnosis and management : a review", J. Am. Med. Assoc, 322, pp.1294~1304 (2019)
- Wang, C., "Anupdateonrenalfibrosis: from mechanisms to therapeutic stra tegies with a focus on extracellular vesicles", Kidney Res. Clin. Pract, 42, pp.174~187 (2023)
- Chuang, P. Y., "Molecular targets for treatment of kidney f ibrosis", J. Mol. Med, 91, pp.549~559 (2013)
- Zhou, Y., "Principles of RNA methylation and their implications for biology and medicine", Biomed. Pharmacother, 131, pp.110731~ (2020)
- Luo, J., "N6-methyladenosine RNA modification in inflammation : roles, mechanisms, and applications", Front. Cell Dev. Biol, 9, pp.670711~ (2021)
- Huang, X., "m6A RNA methylation regulators could contribute to the occurrence of chronic obstructive pulmonary disease", J. Cell Mol. Med, 24, pp.12706~12715 (2020)
- Fan, C., "Comprehensive analysis of the transcriptome-wide m6A methylation modification difference in liver fibrosis mice by high-throughput m6A sequen cing", Front. Cell Dev. Biol, 9, pp.767051~ (2021)
- Yang, L., "New advances of DNA", RNA methylation modification in liver f ibrosis, 2022, pp.92, 110224~
- An, J. N., "Periostin induces kidney fibrosis after acute kidney injury via the p38 MAPK pathway", Am. J. Physiol. Renal Physiol, 316, pp.F426~F437 (2019)
- Varghese, F., "IHC Profiler: an open source plugin for the quantitative evaluation and automated scoring of immunohis tochemistry images of human tissue samples", PLoS ONE, 9, pp.e96801~ (2014)
- Landini, G., "Colour deconvolution : stain unmixing in histological imaging", Bioinformatics, 37, pp.1485~1487 (2021)
- Lv, D., "PDGF signaling inhibits mitophagy in glioblastoma stem cells through N(6)-methyladenosine", Dev. Cell, 57, pp.1466~1481 (2022)
- Lin, S., "Them(6)Amethyltransferase METTL3promotes translation inhuman cancer cells", Mol. Cell, 62, pp.335~345 (2016)
- Andrews, S., "FastQC: A Quality Control Tool for High Throughput Sequence Data" (2010), Babraham Bioinformatics, Babraham Institute
- Bolger, A. M., "Trimmomatic: a flexible trimmer for Illumina sequence data", Bioinformatics, 30, pp.2114~2120 (2014)
- Dobin, A., "STAR : ultrafast universal RNA-seq aligner", Bioinformatics, 29, pp.15~21 (2013)
- Li, B., "RSEM : accurate transcript quantification from RNA-Seq data with or without a reference genome", BMC Bioinforma, 12, pp.323~ (2011)
- Meng, J., "A protocol for RNA methylation differential analysis with MeRIP Seq data and exomePeak R", Bioconductor package, 2014, pp.69, 274~281
- Cui, X., "Guitar : an R", Bioconductor package for gene annotation guided transcriptomic analysis of RNA-related genomic features, 2016, pp.2016, 8367534~
- Heinz, S., "Simple combinations of lineage-determining transcription factors prime cis-regulatory elements required for macrophage and B cell identities", Mol. Cell, 38, pp.576~589 (2010)
- "EnhancedVolcano: publication-ready volcano plots with enhanced colouring and labeling" (2018)[URL]
- Yankova, E., "Small-molecule inhibition of METTL3 as a strategy against myeloid leukaemia", Nature, 593, pp.597~601 (2021)
- Bindea, G., "ClueGO : a Cytoscape plug-in to decipher functionally grouped gene ontology and pathway annotation networks", Bioinformatics, 25, pp.1091~1093 (2009)
- Subramanian, A., "Gene set enrichment analysis : a knowledge-based approach for interpreting genome-wide expression profiles", Proc. Natl. Acad. Sci. USA, 102, pp.15545~15550 (2005)
- Shen, X., "The activity of guanine exchange factor NET1 is essential for transforming growth factor-beta-mediated stress fiber formation", J. Biol. Chem, 276, pp.15362~15368 (2001)
- Lee, J., "Smad3 regulates Rho signaling via NET1 in the transforming growth factor-beta-induced epithelial-mesenchymal transition of human retinal pigment epithelial cells", J. Biol. Chem, 285, pp.26618~26627 (2010)
- Statello, L., "Gene regulation by long non-coding RNAs and its biological functions", Nat. Rev. Mol. Cell Biol, 22, pp.96~118 (2021)
- Jiang, X., "The role of m6A modification in the biological functions and diseases", Signal Transduct. Target Ther, 6, pp.74~ (2021)
- He, P. C., "m(6)A RNA methylation : from mechanisms to therapeutic potential", EMBO J, 40, pp.e105977~ (2021)
- Xiang, Y., "RNA m(6)A methylation regulates the ultraviolet-induced DNA damage response", Nature, 543, pp.573~576 (2017)
- Lin, X., "RNA m(6)A methylation regulates the epithelial mesenchymal tran sition of cancer cells and translation of Snail", Nat. Commun, 10, pp.2065~ (2019)
- Ligresti, G., "Circular RNA methylation : a new twist in lung fibrosis", Am. J. Respir. Cell Mol. Biol, 66, pp.471~472 (2022)
- Wang, S., "The combined effects of circular RNA methylation promote pul monary fibrosis", Am. J. Respir. Cell Mol. Biol, 66, pp.510~523 (2022)
- Liu, P., "m(6)A-induced lncRNA MALAT1 aggravates renal fibrogenesis in obstructive nephropathy through the miR-145", FAK pathway, 2020, pp.12, 5280~5299
- Ning, Y., "Genistein ameliorates renal fibrosis through regulation snail via m6A RNA demethylase ALKBH5", Front. Pharmacol, 11, pp.579265~ (2020)
- Liu, E., "METTL3", N6-methyladenosine, ERK, pp.NF-kappaB pathway activation, J. Cell Mol. Med[DOI]
- Xu, Z., "METTL14-regulated PI3K", Akt signaling pathway via PTEN affects HDAC5-mediated epithelial-mesenchymal transition of renal tubular cells in diabetic kidney disease, 2021, pp.12, 32~
- Meng, F., "METTL3 contributes to renal ischemia-reperfusion injury by reg ulating Foxd1 methylation", Am. J. Physiol. Renal Physiol, 319, pp.F839~F847 (2020)
- Li, C. M., "Alteration of N6-methyladenosine RNA profiles in cisplatin-induced acute kidney injury in mice", Front. Mol. Biosci, 8, pp.654465~ (2021)
- Ramalingam, H., "A methionine-Mettl3-N(6)-methyladenosine axis promotes polycystic kidney disease", Cell. Metab, 33, pp.1234~1247 (2021)