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研究生： 林芮綺
Lin, Ruei-Ci
論文名稱： 探討miR-20b-5p調控滋養層細胞之遷移及侵襲能力之分子機制
Investigate the molecular mechanism of miR-20b-5p in regulating trophoblast migration and invasion
指導教授： 王家義
Wang, Chia-Yih
學位類別： 碩士
Master
系所名稱： 醫學院 - 細胞生物與解剖學研究所
Institute of Cell Biology and Anatomy
論文出版年： 2023
畢業學年度： 111
語文別： 英文
論文頁數： 47
中文關鍵詞： 滋養細胞侵襲 、miR-20b-5p 、ATG16L1 、ATG7 、初級纖毛 、自噬作用
外文關鍵詞： trophoblast invasion, miR-20b-5p, ATG16L1, ATG7, primary cilia, autophagy
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    • 復發性流產疾病被定義為在妊娠 20 週前發生超過 3 次妊娠失敗，此疾病影響大約1%-2%的孕婦且發病原因大多不明。子宮內膜間質分化不良或母胎界面滋養層細胞浸潤缺陷是主要導致復發性流產發生的原因。包括 miR-20b-5p 在內的多種 miRNA 在復發性流產的患者血液中異常表達，然而其導致復發性流產潛在的分子機制仍不清楚。初級纖毛是由微管組成類似天線的細胞器，協調體內發育和分化的信號傳遞。初級纖毛缺陷導致妊娠併發症，如復發性流產或子癇前症。MiR-20b-5p 通過阻斷初級纖毛的形成而抑制滋養細胞浸潤能力。MiR-20b-5p 靶向並抑制 ATG16L1 和 ATG7 表達，從而阻斷自噬作用（Autophagy）。自噬作用（Autophagy）抑制導致初級纖毛的形成減少且降低了初級纖毛下游信號 ERK的激活，ERK信號的激活是滋養細胞浸潤能力中的關鍵信號。阿司匹林在臨床上用於預防復發性流產。阿司匹林治療可抑制 miR-20b-5p 水平，從而促進初級纖毛形成和提升滋養細胞浸潤的能力。總結來說，我們的研究結果說明了 miR-20b-5p 通過降低 ATG16L1 和 ATG7 的表達來抑制初級纖毛形成和滋養細胞浸潤的分子機制，並且阿司匹林治療可以在復發性流產疾病中挽救miR-20b-5p所導致的缺陷表型。

    Recurrent miscarriage is defined as more than three times pregnancy failures occurring before twenty weeks of gestation. Poor differentiation of endometrial stroma or defective trophoblast cell invasion at the maternal-fetal interface leads to recurrent miscarriage. Several miRNAs, including miR-20b-5p, are aberrantly regulated in recurrent miscarriage; however, the underlying molecular mechanism remains unclear. Primary cilium is an antenna like organelle that coordinating signaling for development and differentiation. Defective primary cilia formation leads to pregnant complications such as recurrent miscarriage or preeclampsia. Here we demonstrated that miR-20b-5p inhibited trophoblast cell invasion by blocking primary cilia formation. Mechanistically, miR-20b-5p targeted to and inhibited ATG16L1 and ATG7 expressions thus blocking autophagy. Defective autophagy reduced primary cilia formation followed by impeding ERK activation, a crucial signaling for trophoblast invasion. Aspirin is used to prevent recurrent miscarriage in clinic. Treatment of aspirin inhibited miR-20b-5p levels thus restoring ciliogenesis and trophoblast invasion. Thus, our findings uncover underlying mechanism by which miR-20b-5p suppressed primary cilia formation and trophoblast invasion through reducing the expression of ATG16L1 and ATG7, and the defective phenotypes can be rescued by aspirin in recurrent miscarriage.

    口試合格證明書 I 中文摘要 II ABSTRACT III ACKNOWLEDGEMENT IV CONTENTS VI ABBREVIATIONS VIII INTRODUCTION 1 A.    Molecular mechanism of embryo implantation 1 B.    Function of microRNAs in cell invasion 2 C.    Molecular mechanism of primary cilia 3 D.    Function of autophagy signaling in cell progress 4 MATERIAL AND METHOD 6 RESULTS 12 DISCUSSION 17 FIGURES 20 Figure 1. MiR-155-5p mimic, miR-20b-5p mimic and miR-718 inhibitor have no effect on T-HESC cell growth. 20 Figure 2. Decidualization stimulation in vitro. 21 Figure 3. MiR-20b-5p mimic, miR-155-5p mimic and miR-718 inhibitor have no effect on decidualization. 22 Figure 4. MiR-20b-5p and miR-155-5p inhibit the migration capability of trophoblast. 23 Figure 5. MiR-20b-5p and miR-155-5p inhibit the invasion capability of trophoblast. 24 Figure 6. MiR-155-5p inhibits the cell numbers of trophoblast. 25 Figure 7. MiR-20b-5p inhibits the EMT of trophoblast. 26 Figure 8. MiR-20b-5p inhibits the MMP-9 expression of trophoblast. 27 Figure 9. MiR-20b-5p inhibits the MMP-9 proteolytic activity in trophoblast. 28 Figure 10. Target gene of miR-20b-5p. 29 Figure 11. Knockdown ATG16L1 inhibits trophoblast invasion. 30 Figure 12. MiR-20b-5p blocks autophagy signaling in trophoblast. 31 Figure 13. MiR-20b-5p blocks autophagy signaling to inhibit trophoblast invasion capability. 32 Figure 14. HTR8/SVneo is ciliated cell. 33 Figure 15. MiR-20b-5p inhibits primary cilia of trophoblast. 34 Figure 16. Blocking autophagy inhibits primary cilia. 35 Figure 17. Deficiency of IFT88 inhibits growth of primary cilia. 36 Figure 18. Inhibition of primary cilia reduces trophoblast invasion. 37 Figure 19. MiR-20b-5p reduces phosphorylation of ERK signaling. 38 Figure 20. Aspirin rescues defect of EMT and trophoblast invasion caused by miR-20b-5p. 39 Figure 21. Aspirin rescues inhibition of primary cilia caused by miR-20b-5p. 40 Figure 22. Aspirin treated can not only promotes activation of ERK but inhibits expression of miR-20b-5p. 41 Figure 23. Schematic illustration of underlying mechanism by which miR-20b-5p inhibits trophoblast invasion. 42 REFERENCE 43

    1. You, Y., et al., TNF-alpha Regulated Endometrial Stroma Secretome Promotes Trophoblast Invasion. Front Immunol, 2021. 12: p. 737401.
    2. Zhang, S., et al., Physiological and molecular determinants of embryo implantation. Mol Aspects Med, 2013. 34(5): p. 939-80.
    3. Vento-Tormo, R., et al., Single-cell reconstruction of the early maternal-fetal interface in humans. Nature, 2018. 563(7731): p. 347-353.
    4. Pan-Castillo, B., et al., Morphophysical dynamics of human endometrial cells during decidualization. Nanomedicine, 2018. 14(7): p. 2235-2245.
    5. Gellersen, B. and J. Brosens, Cyclic AMP and progesterone receptor cross-talk in human endometrium: a decidualizing affair. J Endocrinol, 2003. 178(3): p. 357-72.
    6. Daly, D.C., I.A. Maslar, and D.H. Riddick, Prolactin production during in vitro decidualization of proliferative endometrium. Am J Obstet Gynecol, 1983. 145(6): p. 672-8.
    7. Irwin, J.C., et al., Hormonal regulation of human endometrial stromal cells in culture: an in vitro model for decidualization. Fertil Steril, 1989. 52(5): p. 761-8.
    8. Burton, G.J. and E. Jauniaux, What is the placenta? Am J Obstet Gynecol, 2015. 213(4 Suppl): p. S6 e1, S6-8.
    9. Illsley, N.P., et al., Trophoblast invasion: Lessons from abnormally invasive placenta (placenta accreta). Placenta, 2020. 102: p. 61-66.
    10. Meakin, C., E.S. Barrett, and L.M. Aleksunes, Extravillous trophoblast migration and invasion: Impact of environmental chemicals and pharmaceuticals. Reprod Toxicol, 2022. 107: p. 60-68.
    11. Kokkinos, M.I., et al., Cadherins in the human placenta--epithelial-mesenchymal transition (EMT) and placental development. Placenta, 2010. 31(9): p. 747-55.
    12. Serrano-Gomez, S.J., M. Maziveyi, and S.K. Alahari, Regulation of epithelial-mesenchymal transition through epigenetic and post-translational modifications. Mol Cancer, 2016. 15: p. 18.
    13. Zhu, J.Y., Z.J. Pang, and Y.H. Yu, Regulation of trophoblast invasion: the role of matrix metalloproteinases. Rev Obstet Gynecol, 2012. 5(3-4): p. e137-43.
    14. Wang, X.H., et al., Low chorionic villous succinate accumulation associates with recurrent spontaneous abortion risk. Nat Commun, 2021. 12(1): p. 3428.
    15. Chen, C.H., et al., A novel platform for discovery of differentially expressed microRNAs in patients with repeated implantation failure. Fertil Steril, 2021. 116(1): p. 181-188.
    16. Ambros, V., The functions of animal microRNAs. Nature, 2004. 431(7006): p. 350-5.
    17. Basavarajappa, D., et al., Dicer up-regulation by inhibition of specific proteolysis in differentiating monocytic cells. Proc Natl Acad Sci U S A, 2020. 117(15): p. 8573-8583.
    18. Sun, Z., et al., Effect of exosomal miRNA on cancer biology and clinical applications. Mol Cancer, 2018. 17(1): p. 147.
    19. Muys, B.R., et al., miR-450a Acts as a Tumor Suppressor in Ovarian Cancer by Regulating Energy Metabolism. Cancer Res, 2019. 79(13): p. 3294-3305.
    20. Singh, R., et al., MicroRNA-195 inhibits proliferation, invasion and metastasis in breast cancer cells by targeting FASN, HMGCR, ACACA and CYP27B1. Sci Rep, 2015. 5: p. 17454.
    21. Skalis, G., et al., MicroRNAs in Preeclampsia. Microrna, 2019. 8(1): p. 28-35.
    22. Zhang, H., et al., miR-30-5p-mediated ferroptosis of trophoblasts is implicated in the pathogenesis of preeclampsia. Redox Biol, 2020. 29: p. 101402.
    23. Wang, C.Y., et al., Elevated miR-200a and miR-141 inhibit endocrine gland-derived vascular endothelial growth factor expression and ciliogenesis in preeclampsia. J Physiol, 2019. 597(12): p. 3069-3083.
    24. Wang, C.Y., et al., Primary Cilium-Regulated EG-VEGF Signaling Facilitates Trophoblast Invasion. J Cell Physiol, 2017. 232(6): p. 1467-1477.
    25. Yamamoto, Y. and N. Mizushima, Autophagy and Ciliogenesis. JMA J, 2021. 4(3): p. 207-215.
    26. Anvarian, Z., et al., Cellular signalling by primary cilia in development, organ function and disease. Nat Rev Nephrol, 2019. 15(4): p. 199-219.
    27. Chen, Z., et al., CP110, a cell cycle-dependent CDK substrate, regulates centrosome duplication in human cells. Dev Cell, 2002. 3(3): p. 339-50.
    28. Schmidt, T.I., et al., Control of centriole length by CPAP and CP110. Curr Biol, 2009. 19(12): p. 1005-11.
    29. Goetz, S.C., K.F. Liem, Jr., and K.V. Anderson, The spinocerebellar ataxia-associated gene Tau tubulin kinase 2 controls the initiation of ciliogenesis. Cell, 2012. 151(4): p. 847-858.
    30. Brown, J.M. and G.B. Witman, Cilia and Diseases. Bioscience, 2014. 64(12): p. 1126-1137.
    31. Kenny, T.D. and P.L. Beales, Ciliopathies: A reference for clinicians. 2013: Oxford University Press.
    32. Szenker-Ravi, E., et al., Discovery of a genetic module essential for assigning left-right asymmetry in humans and ancestral vertebrates. Nat Genet, 2022. 54(1): p. 62-72.
    33. He, Y., et al., Abnormal activation of the sonic hedgehog signaling pathway in endometriosis and its diagnostic potency. Fertil Steril, 2018. 110(1): p. 128-136 e2.
    34. Matsumoto, H., et al., Indian hedgehog as a progesterone-responsive factor mediating epithelial-mesenchymal interactions in the mouse uterus. Dev Biol, 2002. 245(2): p. 280-90.
    35. Lee, K., et al., Indian hedgehog is a major mediator of progesterone signaling in the mouse uterus. Nat Genet, 2006. 38(10): p. 1204-9.
    36. Nygaard, M.B., et al., Cell context-specific expression of primary cilia in the human testis and ciliary coordination of Hedgehog signalling in mouse Leydig cells. Sci Rep, 2015. 5: p. 10364.
    37. Sedkov, Y., et al., Methylation at lysine 4 of histone H3 in ecdysone-dependent development of Drosophila. Nature, 2003. 426(6962): p. 78-83.
    38. Tsai, Y.C., et al., PDGF-AA activates AKT and ERK signaling for testicular interstitial Leydig cell growth via primary cilia. J Cell Biochem, 2023. 124(1): p. 89-102.
    39. Mizushima, N. and M. Komatsu, Autophagy: renovation of cells and tissues. Cell, 2011. 147(4): p. 728-41.
    40. Morleo, M. and B. Franco, The Autophagy-Cilia Axis: An Intricate Relationship. Cells, 2019. 8(8).
    41. Ferrante, M.I., et al., Oral-facial-digital type I protein is required for primary cilia formation and left-right axis specification. Nat Genet, 2006. 38(1): p. 112-7.
    42. Tang, Z., et al., Autophagy promotes primary ciliogenesis by removing OFD1 from centriolar satellites. Nature, 2013. 502(7470): p. 254-7.
    43. Zhu, J., et al., ATG7 Promotes Bladder Cancer Invasion via Autophagy-Mediated Increased ARHGDIB mRNA Stability. Adv Sci (Weinh), 2019. 6(8): p. 1801927.
    44. Nakashima, A., et al., Impaired autophagy by soluble endoglin, under physiological hypoxia in early pregnant period, is involved in poor placentation in preeclampsia. Autophagy, 2013. 9(3): p. 303-16.
    45. Staun-Ram, E., et al., Expression and importance of matrix metalloproteinase 2 and 9 (MMP-2 and -9) in human trophoblast invasion. Reprod Biol Endocrinol, 2004. 2: p. 59.
    46. Peng, Y.F., et al., Autophagy inhibition suppresses pulmonary metastasis of HCC in mice via impairing anoikis resistance and colonization of HCC cells. Autophagy, 2013. 9(12): p. 2056-68.
    47. Han, J.H., et al., Snail acetylation by autophagy-derived acetyl-coenzyme A promotes invasion and metastasis of KRAS-LKB1 co-mutated lung cancer cells. Cancer Commun (Lond), 2022. 42(8): p. 716-749.
    48. Wang, M., et al., Purple sweet potato delphinidin-3-rutin represses glioma proliferation by inducing miR-20b-5p/Atg7-dependent cytostatic autophagy. Mol Ther Oncolytics, 2022. 26: p. 314-329.
    49. Borovina, A. and B. Ciruna, IFT88 plays a cilia- and PCP-independent role in controlling oriented cell divisions during vertebrate embryonic development. Cell Rep, 2013. 5(1): p. 37-43.
    50. de Jong, P.G., et al., Aspirin and/or heparin for women with unexplained recurrent miscarriage with or without inherited thrombophilia. Cochrane Database Syst Rev, 2014. 2014(7): p. Cd004734.
    51. Su, M.T., et al., Aspirin enhances trophoblast invasion and represses soluble fms-like tyrosine kinase 1 production: a putative mechanism for preventing preeclampsia. J Hypertens, 2019. 37(12): p. 2461-2469.
    52. Xu, Y., et al., Let-7i-Induced Atg4B Suppression Is Essential for Autophagy of Placental Trophoblast in Preeclampsia. J Cell Physiol, 2017. 232(9): p. 2581-2589.
    53. Cao, B., et al., Autophagy regulation of physiological and pathological processes in the female reproductive tract. Am J Reprod Immunol, 2017. 77(5).
    54. Tsukamoto, S., et al., Autophagy is essential for preimplantation development of mouse embryos. Science, 2008. 321(5885): p. 117-20.
    55. Kuma, A., et al., The role of autophagy during the early neonatal starvation period. Nature, 2004. 432(7020): p. 1032-6.
    56. Tan, H.X., et al., Autophagy suppression of trophoblast cells induces pregnancy loss by activating decidual NK cytotoxicity and inhibiting trophoblast invasion. Cell Commun Signal, 2020. 18(1): p. 73.
    57. Hassan, E., et al., Abnormal ciliogenesis in decidual stromal cells in recurrent miscarriage. J Reprod Immunol, 2022. 150: p. 103486.
    58. Zhang, X., et al., Circular RNA circNRIP1 acts as a microRNA-149-5p sponge to promote gastric cancer progression via the AKT1/mTOR pathway. Mol Cancer, 2019. 18(1): p. 20.
    59. Yuan, J., et al., MicroRNA-7 inhibits hepatocellular carcinoma cell invasion and metastasis by regulating Atg5-mediated autophagy. Transl Cancer Res, 2020. 9(6): p. 3965-3972.
    60. Qiu, Z., et al., CircHIPK3 regulates the autophagy and apoptosis of hypoxia/reoxygenation-stimulated cardiomyocytes via the miR-20b-5p/ATG7 axis. Cell Death Discov, 2021. 7(1): p. 64.
    61. Cao, B., C. Macones, and I.U. Mysorekar, ATG16L1 governs placental infection risk and preterm birth in mice and women. JCI Insight, 2016. 1(21): p. e86654.
    62. Burton, G.J., et al., Rheological and physiological consequences of conversion of the maternal spiral arteries for uteroplacental blood flow during human pregnancy. Placenta, 2009. 30(6): p. 473-82.
    63. Yang, H., et al., miR-20b-5p functions as tumor suppressor microRNA by targeting cyclinD1 in colon cancer. Cell Cycle, 2020. 19(21): p. 2939-2954.
    64. Paseban, M., et al., Modulation of microRNAs by aspirin in cardiovascular disease. Trends Cardiovasc Med, 2020. 30(5): p. 249-254.
    65. Kunzmann, A.T., et al., PTGS2 (Cyclooxygenase-2) expression and survival among colorectal cancer patients: a systematic review. Cancer Epidemiol Biomarkers Prev, 2013. 22(9): p. 1490-7.
    66. Wang, D. and R.N. Dubois, Prostaglandins and cancer. Gut, 2006. 55(1): p. 115-22.
    67. Moffett, A. and C. Loke, Immunology of placentation in eutherian mammals. Nat Rev Immunol, 2006. 6(8): p. 584-94.
    68. Kingdom, J.C., et al., A placenta clinic approach to the diagnosis and management of fetal growth restriction. Am J Obstet Gynecol, 2018. 218(2S): p. S803-S817.
    69. Collins, S.L., et al., Measurement of spiral artery jets: general principles and differences observed in small-for-gestational-age pregnancies. Ultrasound Obstet Gynecol, 2012. 40(2): p. 171-8.

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