{"paper_id":"959371e0-d396-4b0a-9d8c-9760664789fe","body_text":"Critical Reviews™ in Immunology\nPublished 6 issues per year\nISSN Print: 1040-8401\nISSN Online: 2162-6472\nResearch Article\nNLRC3 Inhibits the Migration and Invasion of Adenomyosis by Modulating the PI3K/AKT/mTOR Pathway in Endometrial Cells\nABSTRACT\nQuantitative real-time PCR (qRT-PCR), western blotting, and immunohistochemical (IHC) staining were used to assess the expression of NLRC3 in tissues and cells. The effects of NLRC3 on the proliferation, apoptosis, migration, and invasion of endometrial cells were investigated via Cell Counting Kit-8 (CCK-8), colony formation, 5-Ethynyl-2'-deoxyuridine (EdU), flow cytometry, cell scratch and transwell assays, respectively. The mouse model of adenomyosis was constructed. The regulation mechanisms by which NLRC3 acts were further verified in vivo study. The study revealed epithelial-mesenchymal transition (EMT) related protein expression was upregulated and NLRC3 was downregulated in endometria of patients with adenomyosis. Upregulation of NLRC3 expression reduced endometrial cell growth, migration, invasion, and promoted cell apoptosis rate. Mechanistically, upregulation of NLRC3 expression inhibited the level of EMT and blocked the PI3K/AKT/mTOR pathway activation in endometrial cells. In vivo, increased the expression of NLRC3 decreased the levels of cytokines (IL-6 and IL-8), inhibited the levels of PI3K/AKT/mTOR pathway related genes and mitigated disease severity. Our findings indicate that NLRC3 inhibits migration and invasion of adenomyosis by modulating PI3K/AKT/mTOR pathway in endometrial cells. Consequently, NLRC3 holds promise as a potential therapeutic target for adenomyosis management.\nFigures\n-\nStruble J, Reid S, Bedaiwy MA. Adenomyosis: A clinical review of a challenging gynecologic condition. J Minim Invasive Gynecol. 2016;23(2):164-85.\n-\nHashimoto A, Iriyama T, Sayama S, Okamura A, Kato K, Fujii T, Kubota K, Ichinose M, Sone K, Kumasawa K, Nagamatsu T, Hirota Y, Osuga Y. Differences in the incidence of obstetric complications depending on the extent and location of adenomyosis lesions. J Matern Fetal Neonatal Med. 2023;36(2):2226789.\n-\nBourdon M, Santulli P, Oliveira J, Marcellin L, Maignien C, Melka L, Bordonne C, Millisher AE, Plu-Bureau G, Cormier J, Chapron C. Focal adenomyosis is associated with primary infertility. Fertil Steril. 2020;114(6):1271-7.\n-\nZhai J, Vannuccini S, Petraglia F, Giudice LC. Adenomyosis: Mechanisms and pathogenesis. Semin Reprod Med. 2020;38(2-3):129-43.\n-\nRees CO, Rupert IAM, Nederend J, Consten D, Mischi M, van Vliet H AAM, Schoot BC. Women with combined adenomyosis and endometriosis on MRI have worse IVF/ICSI outcomes compared to adenomyosis and endometriosis alone: A matched retrospective cohort study. Eur J Obstet Gynecol Reprod Biol. 2022;271:223-34.\n-\nYang YT, Jiang XY, Xu HL, Chen G, Wang SL, Zhang HP, Hong L, Jin QQ, Yao H, Zhang WY, Zhu YT, Mei J, Tian L, Ying J, Hu JJ, Zhou SG. Autoimmune disease-related hub genes are potential biomarkers and associated with immune microenvironment in endometriosis. Int J Gen Med. 2023;16:2897-921.\n-\nBourdon M, Santulli P, Jeljeli M, Vannuccini S, Marcellin L, Doridot L, Petraglia F, Batteux F, Chapron C. Immunological changes associated with adenomyosis: A systematic review. Hum Reprod Update. 2021;27(1):108-29.\n-\nNakashima M, Suga N, Ikeda Y, Yoshikawa S, Matsuda S. Relevant microRNAs of MMPs and TIMPs with certain gut microbiota could be involved in the invasiveness and metastasis of malignant tumors. Innov Discov. 2024;1(2):10.\n-\nWynn TA, Vannella KM. Macrophages in tissue repair, regeneration, and fibrosis. Immunity. 2016;44(3):450-62.\n-\nKaunain SF, Pandurangan AK. Immunotherapy for colorectal cancer: Recent advancements. Innov Discov. 2025; 2(3):12.\n-\nSchneider M, Zimmermann AG, Roberts RA, Zhang L, Swanson KV, Wen H, Davis BK, Allen IC, Holl EK, Ye Z, Rahman AH, Conti BJ, Eitas TK, Koller BH, Ting JP. The innate immune sensor NLRC3 attenuates Toll-like receptor signaling via modification of the signaling adaptor TRAF6 and transcription factor NF-kB. Nat Immunol. 2012;13(9):823-31.\n-\nZhu H, Cao X. NLR members in inflammation-associated carcinogenesis. Cell Mol Immunol. 2017;14(5):403-5.\n-\nHu S, Du X, Huang Y, Fu Y, Yang Y, Zhan X, He W, Wen Q, Zhou X, Zhou C, Zhong XP, Yang J, Xiong W, Wang R, Gao Y, Ma L. NLRC3 negatively regulates CD4+ T cells and impacts protective immunity during Mycobacterium tuberculosis infection. PLoS Pathog. 2018;14(8):e1007266.\n-\nZhang L, Mo J, Swanson KV, Wen H, Petrucelli A, Gregory SM, Zhang Z, Schneider M, Jiang Y, Fitzgerald KA, Ouyang S, Liu ZJ, Damania B, Shu HB, Duncan JA, Ting JP. NLRC3, a member of the NLR family of proteins, is a negative regulator of innate immune signaling induced by the DNA sensor STING. Immunity. 2014;40(3):329-41.\n-\nKarki R, Man SM, Malireddi RKS, Kesavardhana S, Zhu Q, Burton AR, Sharma BR, Qi X, Pelletier S, Vogel P, Rosenstiel P, Kanneganti TD. NLRC3 is an inhibitory sensor of PI3K-mTOR pathways in cancer. Nature. 2016;540(7634):583-7.\n-\nLi X, Deng M, Petrucelli AS, Zhu C, Mo J, Zhang L, Tam JW, Ariel P, Zhao B, Zhang S, Ke H, Li P, Dokholyan NV, Duncan JA, Ting JP. Viral DNA binding to NLRC3, an inhibitory nucleic acid sensor, unleashes STING, a cyclic dinucleotide receptor that activates type I interferon. Immunity. 2019;50(3):591-9.e6.\n-\nAn M, Fu X, Meng X, Liu H, Ma Y, Li Y, Li Q, Chen J. PI3K/AKT signaling pathway associates with pyroptosis and inflammation in patients with endometriosis. J Reprod Immunol. 2024;162:104213.\n-\nMao X, Wang Y, Carter AV, Zhen X, Guo SW. The retardation of myometrial infiltration, reduction of uterine contractility, and alleviation of generalized hyperalgesia in mice with induced adenomyosis by levo-tetrahydropalmatine (l-THP) and andrographolide. Reprod Sci. 2011;18(10):1025-37.\n-\nAcloque H, Adams MS, Fishwick K, Bronner-Fraser M, Nieto MA. Epithelial-mesenchymal transitions: The importance of changing cell state in development and disease. J Clin Invest. 2009;119(6):1438-49.\n-\nShi X, Wang J, Lei Y, Cong C, Tan D, Zhou X. Research progress on the PI3K/AKT signaling pathway in gynecological cancer. Mol Med Rep. 2019;19(6):4529-35.\n-\nKay N, Huang CY, Shiu LY, Yu YC, Chang Y, Schatz F, Suen JL, Tsai EM, Huang SJ. TGF-B1 neutralization improves pregnancy outcomes by restoring endometrial receptivity in mice with adenomyosis. Reprod Sci. 2021;28(3):877-87.\n-\nHogg C, Horne AW, Greaves E. Endometriosis-associated macrophages: Origin, phenotype, and function. Front Endocrinol. 2020;11:7.\n-\nWang Y, Nicholes K, Shih IM. The origin and pathogenesis of endometriosis. Annu Rev Pathol. 2020;15:71-95.\n-\nOhto U. Activation and regulation mechanisms of NOD-like receptors based on structural biology. Front Immunol. 2022;13:953530.\n-\nZhou JT, Ren KD, Hou J, Chen J, Yang G. a‑rhamnrtin‑3‑a‑rhamnoside exerts anti‑inflammatory effects on lipopolysaccharide‑stimulated RAW264.7 cells by abrogating NF‑κB and activating the Nrf2 signaling pathway. Mol Med Rep. 2021;24(5):799.\n-\nFu Y, Zhan X, Wang Y, Jiang X, Liu M, Yang Y, Huang Y, Du X, Zhong XP, Li L, Ma L, Hu S. NLRC3 expression in dendritic cells attenuates CD4(+) T cell response and autoimmunity. EMBO J. 2019;38(16):e101397.\n-\nLi ZT, Liu H, Zhang WQ. NLRC3 alleviates hypoxia/reoxygenation induced inflammation in RAW264.7 cells by inhibiting K63-linked ubiquitination of TRAF6. Hepatobiliary Pancreat Dis Int. 2020;19(5):455-60.\n-\nTaki M, Abiko K, Ukita M, Murakami R, Yamanoi K, Yamaguchi K, Hamanishi J, Baba T, Matsumura N, Mandai M. Tumor immune microenvironment during epithelial-mesenchymal transition. Clin Cancer Res. 2021;27(17):4669-79.\n-\nGrund EM, Kagan D, Tran CA, Zeitvogel A, Starzinski-Powitz A, Nataraja S, Palmer SS. Tumor necrosis factor-alpha regulates inflammatory and mesenchymal responses via mitogen-activated protein kinase kinase, p38, and nuclear factor kappaB in human endometriotic epithelial cells. Mol Pharmacol. 2008;73(5):1394-404.\n-\nUchimura T, Oyama Y, Deng M, Guo H, Wilson JE, Rampanelli E, Cook KD, Misumi I, Tan X, Chen L, Johnson B, Tam J, Chou WC, Brickey WJ, Petrucelli A, Whitmire JK, Ting JPY. The innate immune sensor NLRC3 acts as a rheostat that fine-tunes T cell responses in infection and autoimmunity. Immunity. 2018;49(6):1049-61.e6.\n-\nZhuo Y, Li X, Feng W. NLRC3 is a potential prognostic biomarker that is correlated with immune cell infiltration in lung adenocarcinoma. Sci Rep. 2023;13(1):2923.\n-\nLi R, Zhao Y, Zhang X, Yang L, Zou X. NLRC3 participates in inhibiting the pulmonary inflammatory response of sepsis-induced acute lung injury. Immunol Invest. 2023;52(5):567-82.\nSenior Associate Scientist, Prime Therapeutics, USA","source_license":"CC0","license_restricted":false}