Abstract
Chronic glomerulonephritis (CGN) is a leading cause of end-stage renal disease in China; thus, there is an urgent need for effective therapeutic targets and strategies for CGN treatment. However, studies on CGN pathogenesis are limited. In this study, we found that the fat mass and obesity-associated protein (FTO) was significantly decreased in the lipopolysaccharide (LPS)-induced human glomerular mesangial cells (HGMCs) (P < 0.01) and kidney tissues of CGN patients (P < 0.05). Moreover, double-labeling immunofluorescence and flow cytometry assays demonstrated that the overexpression of FTO could inhibit inflammation and excessive proliferation of HGMCs. Furthermore, RNA-sequencing (RNA-seq) and real-time quantitative polymerase chain reaction (RT-qPCR) analyses revealed that FTO overexpression induced differential expression of 269 genes (absolute fold change ≥ 2 and P-value < 0.05), including 143 upregulated and 126 downregulated genes. Further functional analysis of these differentially expressed genes by Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses suggested that FTO possibly mediates its inhibitory function by regulating the mammalian target of rapamycin (mTOR) signaling pathway and substance metabolism. Lastly, analysis of the PPI network and further identification of the top 10 hub genes (RPS15, RPS18, RPL18A, GNB2L1, RPL19, EEF1A1, RPS25, FAU, UBA52, and RPS6) indicated that FTO mediates its function by affecting the ribosomal proteins. Therefore, in this study, we elucidated the important role of FTO in the regulation of inflammation and excessive proliferation of HGMCs, suggesting FTO administration as a suitable therapeutic intervention for CGN.
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The datasets supporting the conclusions of this article can be obtained from the corresponding author upon reasonable request.
References
AlYousef A, AlSahow A, AlHelal B, Alqallaf A, Abdallah E, Abdellatif M, Nawar H, Elmahalawy R (2020) Glomerulonephritis histopathological pattern change. BMC Nephrol 21(1):186. https://doi.org/10.1186/s12882-020-01836-3
Avraham S, Korin B, Chung JJ, Oxburgh L, Shaw AS (2021) The mesangial cell - the glomerular stromal cell. Nat Rev Nephrol 17(12):855–864. https://doi.org/10.1038/s41581-021-00474-8
Bakshi A, Moin M, Gayatri MB, Reddy ABM, Datla R, Madhav MS, Kirti PB (2023) Involvement of target of rapamycin (TOR) signaling in the regulation of crosstalk between ribosomal protein small subunit 6 kinase-1 (RPS6K-1) and ribosomal proteins. Plants (basel) 12(1):176. https://doi.org/10.3390/plants12010176
Barba C, Benoit B, Bres E, Chanon S, Vieille-Marchiset A, Pinteur C, Pesenti S, Glorieux G, Picard C, Fouque D, Soulage CO, Koppe L (2021) A low aromatic amino-acid diet improves renal function and prevent kidney fibrosis in mice with chronic kidney disease. Sci Rep 11(1):19184. https://doi.org/10.1038/s41598-021-98718-x
de Soysa AKH, Langaas M, Jakic A, Shojaee-Moradie F, Umpleby AM, Grill V, Mostad IL (2021) The fat mass and obesity-associated (FTO) gene allele rs9939609 and glucose tolerance, hepatic and total insulin sensitivity, in adults with obesity. PloS One 16(3):e0248247. https://doi.org/10.1371/journal.pone.0248247
Feng FZ, Gong WY, Li SY, Deng YH (2022) Inhibitory effect of resveratrol on LPS-induced glomerular mesangial cells proliferation and TGF-β1 expression via sphingosine kinase 1 pathway. Chin J Integr Med 29(6):500–507. https://doi.org/10.1007/s11655-022-3530-y
Gao JR, Shi MM, Jiang H, Zhu XL, Wei LB, Qin XJ (2022) MicroRNA-339-5p inhibits lipopolysaccharide-induced rat mesangial cells by regulating the Syk/Ras/c-Fos pathway. Naunyn Schmiedebergs Arch Pharmacol 395(9):1075–1085. https://doi.org/10.1007/s00210-022-02261-z
He Z, Houghton PJ, Williams TM, Shen C (2021) Regulation of DNA duplication by the mTOR signaling pathway. Cell Cycle (Georgetown, Tex) 20(8):742–751. https://doi.org/10.1080/15384101.2021.1897271
Huo YB, Gao X, Peng Q, Nie Q, Bi W (2022) Dihydroartemisinin alleviates AngII-induced vascular smooth muscle cell proliferation and inflammatory response by blocking the FTO/NR4A3 axis. Inflamm Res 71(2):243–253. https://doi.org/10.1007/s00011-021-01533-3
Kisly I, Tamm T (2023) Archaea/eukaryote-specific ribosomal proteins - guardians of a complex structure. Comput Struct Biotechnol J 21:1249–1261. https://doi.org/10.1016/j.csbj.2023.01.037
Li X, Li Y, Wang Y, He X (2022) The m6A demethylase FTO promotes renal epithelial-mesenchymal transition by reducing the m6A modification of lncRNA GAS5. Cytokine 159:156000. https://doi.org/10.1016/j.cyto.2022.156000
Liu H, Li W, He Q, Xue J, Wang J, Xiong C, Pu X, Nie Z (2017) Mass spectrometry imaging of kidney tissue sections of rat subjected to unilateral ureteral obstruction. Sci Rep 7:41954. https://doi.org/10.1038/srep41954
Liu B, Lin J, Bai L, Zhou Y, Lu R, Zhang P, Chen D, Li H, Song J, Liu X, Wu Y, Wu J, Liang C, Zhou J (2019) Paeoniflorin inhibits mesangial cell proliferation and inflammatory response in rats with mesangial proliferative glomerulonephritis through PI3K/AKT/GSK-3β pathway. Front Pharmacol 10:978. https://doi.org/10.3389/fphar.2019.00978
Liu XY, Wang YH, Wang J, Quan JK, Li XD, Guan KP (2023) The role of CSE1L silencing in the regulation of proliferation and apoptosis via the AMPK/mTOR signaling pathway in chronic myeloid leukemia. Hematology 28(1):1–9. https://doi.org/10.1080/16078454.2022.2161201
Miao X, Xiang Y, Mao W, Chen Y, Li Q, Fan B (2020) TRIM27 promotes IL-6-induced proliferation and inflammation factor production by activating STAT3 signaling in HaCaT cells. Am J Physiol Cell Physiol 318(2):C272–C281. https://doi.org/10.1152/ajpcell.00314.2019
Øvrehus MA, Bruheim P, Ju W, Zelnick LR, Langlo KA, Sharma K, de Boer IH, Hallan SI (2018) Gene expression studies and targeted metabolomics reveal disturbed serine, methionine, and tyrosine metabolism in early hypertensive nephrosclerosis. Kidney Int Rep 4(2):321–333. https://doi.org/10.1016/j.ekir.2018.10.007
Shen J, Wu Q, Liang T, Zhang J, Bai J, Yuan M, Shen P (2021) TRIM40 inhibits IgA1-induced proliferation of glomerular mesangial cells by inactivating NLRP3 inflammasome through ubiquitination. Mol Immunol 140:225–232. https://doi.org/10.1016/j.molimm.2021.10.012
Sun PF, Tian T, Chen LN, Fu RG, Xu SS, Ai H, Wang B, Zhang J, Si RY, Chai Z, Cooper ME, Ren ST (2018) Ultrasound combined with microbubbles enhances the effects of methylprednisolone in lipopolysaccharide-induced human mesangial cells. J Pharmacol Exp Ther 365(3):476–484. https://doi.org/10.1124/jpet.117.246223
Sun Q, Geng H, Zhao M, Li Y, Chen X, Sha Q, Lai P, Tang D, Yang D, Liang J, Guo M (2022) FTO-mediated m6 A modification of SOCS1 mRNA promotes the progression of diabetic kidney disease. Clin Transl Med 12(6):e942. https://doi.org/10.1002/ctm2.942
Tsai KD, Lee WX, Chen W, Chen BY, Chen KL, Hsiao TC, Wang SH, Lee YJ, Liang SY, Shieh JC, Lin TH (2018) Upregulation of PRMT6 by LPS suppresses klotho expression through interaction with NF-κB in glomerular mesangial cells. J Cell Biochem 119(4):3404–3416. https://doi.org/10.1002/jcb.26511
Wang JN, Wang F, Ke J, Li Z, Xu CH, Yang Q, Chen X, He XY, He Y, Suo XG, Li C, Yu JT, Jiang L, Ni WJ, Jin J, Liu MM, Shao W, Yang C, Gong Q, Chen HY, Meng XM (2022) Inhibition of METTL3 attenuates renal injury and inflammation by alleviating TAB3 m6A modifications via IGF2BP2-dependent mechanisms. Sci Transl Med 14(640):eabk2709. https://doi.org/10.1126/scitranslmed.abk2709
Xue JF, Shi ZM, Zou J, Li XL (2017) Inhibition of PI3K/AKT/mTOR signaling pathway promotes autophagy of articular chondrocytes and attenuates inflammatory response in rats with osteoarthritis. Biomed Pharmacother 89:1252–1261. https://doi.org/10.1016/j.biopha.2017.01.130
Yang C, Wang H, Zhao X, Matsushita K, Coresh J, Zhang L, Zhao MH (2020) CKD in China: evolving spectrum and public health implications. Am J Kidney Dis 76(2):258–264. https://doi.org/10.1053/j.ajkd.2019.05.032
Yang W, Xie L, Wang P, Zhuang C (2022) MiR-155 regulates m6A level and cell progression by targeting FTO in clear cell renal cell carcinoma. Cell Signal 91:110217. https://doi.org/10.1016/j.cellsig.2021.110217
Yang Y, Li Q, Ling Y, Leng L, Ma Y, Xue L, Lu G, Ding Y, Li J, Tao S (2023) m6A eraser FTO modulates autophagy by targeting SQSTM1/P62 in the prevention of canagliflozin against renal fibrosis. Front Immunol 13:1094556. https://doi.org/10.3389/fimmu.2022.1094556
Yu JT, Hu XW, Chen HY, Yang Q, Li HD, Dong YH, Zhang Y, Wang JN, Jin J, Wu YG, Li J, Ge JF, Meng XM (2021) DNA methylation of FTO promotes renal inflammation by enhancing m6A of PPAR-α in alcohol-induced kidney injury. Pharmacol Res 163:105286. https://doi.org/10.1016/j.phrs.2020.105286
Yuan Y, Park J, Feng A, Awasthi P, Wang Z, Chen Q, Iglesias-Bartolome R (2020) YAP1/TAZ-TEAD transcriptional networks maintain skin homeostasis by regulating cell proliferation and limiting KLF4 activity. Nat Commun 11(1):1472. https://doi.org/10.1038/s41467-020-15301-0
Zhang L, Long J, Jiang W, Shi Y, He X, Zhou Z, Li Y, Yeung RO, Wang J, Matsushita K, Coresh J, Zhao MH, Wang H (2016) Trends in chronic kidney disease in China. N Engl J Med 375(9):905–906. https://doi.org/10.1056/NEJMc1602469
Zhou Y, Hambly BD, McLachlan CS (2017) FTO associations with obesity and telomere length. J Biomed Sci 24(1):65. https://doi.org/10.1186/s12929-017-0372-6
Funding
This study was financially supported by the National Natural Science Foundation of China (No. 81973546) and the Key Scientific Research Projects of Natural Science in Colleges and Universities in Anhui Province (No. 2022AH050747).
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JG and XZ conceived and designed the experiment. TL conducted the experiments and collected data. YG and LW interpreted and analyzed the sequencing data. XZ drafted the original manuscript. JG revised the manuscript. The authors declare that the study data were generated in-house and that no paper mill was used. All the authors approved the final manuscript for submission.
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This study was approved by the Ethics Committee of the Chaohu Hospital of Anhui Medical University (Anhui, China) (KYXM-202208–006). All the patients provided written informed consent.
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Zhuang, X., Liu, T., Wei, L. et al. RNA sequencing reveals the mechanism of FTO in inhibiting inflammation and excessive proliferation of lipopolysaccharide-induced human glomerular mesangial cells. Naunyn-Schmiedeberg's Arch Pharmacol 396, 3835–3846 (2023). https://doi.org/10.1007/s00210-023-02570-x
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DOI: https://doi.org/10.1007/s00210-023-02570-x