Advertisement

The molecular mechanism of miR-96-5p in the pathogenesis and treatment of polycystic ovary syndrome

  • Author Footnotes
    # Co-first authors: Liu Yajing, Zhang Shanshan, Li Chen contributed equally to this work.
    Yajing Liu
    Footnotes
    # Co-first authors: Liu Yajing, Zhang Shanshan, Li Chen contributed equally to this work.
    Affiliations
    State Key Laboratory of Agrobiotechnology, College of Biological Sciences, China Agricultural University, Beijing, China
    Search for articles by this author
  • Author Footnotes
    # Co-first authors: Liu Yajing, Zhang Shanshan, Li Chen contributed equally to this work.
    Shanshan Zhang
    Footnotes
    # Co-first authors: Liu Yajing, Zhang Shanshan, Li Chen contributed equally to this work.
    Affiliations
    State Key Laboratory of Agrobiotechnology, College of Biological Sciences, China Agricultural University, Beijing, China
    Search for articles by this author
  • Author Footnotes
    # Co-first authors: Liu Yajing, Zhang Shanshan, Li Chen contributed equally to this work.
    Li Chen
    Footnotes
    # Co-first authors: Liu Yajing, Zhang Shanshan, Li Chen contributed equally to this work.
    Affiliations
    Department of Reproductive Medicine, Affiliated Jinling Hospital, Medicine School of Nanjing University, Nanjing, Jiangsu, China
    Search for articles by this author
  • Xuan Huang
    Affiliations
    Department of Reproductive Medicine, Affiliated Jinling Hospital, Medicine School of Nanjing University, Nanjing, Jiangsu, China
    Search for articles by this author
  • Mingming Wang
    Affiliations
    Xuzhou Medical University, School of Basic Medical Sciences, Xuzhou, Jiangsu, China
    Search for articles by this author
  • Donata Ponikwicka-Tyszko
    Affiliations
    Department of Biology and Pathology of Human Reproduction, Institute of Animal Reproduction and Food Research, Polish Academy of Sciences, Olsztyn, Poland

    Institute of Biomedicine, Faculty of Medicine, University of Turku, Turku, Finland
    Search for articles by this author
  • Nafis A. Rahman
    Affiliations
    Institute of Biomedicine, Faculty of Medicine, University of Turku, Turku, Finland

    Department of Reproduction and Gynecological Endocrinology, Medical University of Bialystok, Bialystok, Poland
    Search for articles by this author
  • Slawomir Wolczynski
    Affiliations
    Department of Reproduction and Gynecological Endocrinology, Medical University of Bialystok, Bialystok, Poland
    Search for articles by this author
  • Bing Yao
    Correspondence
    Reprint requests: Bing Yao, Department of Reproductive Medicine, Affiliated Jinling Hospital, Medicine School of Nanjing University, Nanjing, Jiangsu 210002, China.
    Affiliations
    Department of Reproductive Medicine, Affiliated Jinling Hospital, Medicine School of Nanjing University, Nanjing, Jiangsu, China
    Search for articles by this author
  • Xiangdong Li
    Correspondence
    Reprint requests: Xiangdong Li, State Key Laboratory of Agrobiotechnology, College of Biological Sciences, China Agricultural University, Beijing 100193, China.
    Affiliations
    State Key Laboratory of Agrobiotechnology, College of Biological Sciences, China Agricultural University, Beijing, China

    Department of Reproduction and Gynecological Endocrinology, Medical University of Bialystok, Bialystok, Poland

    Hainan Sanya Research Institute, Seed Laboratory, Sanya, China
    Search for articles by this author
  • Author Footnotes
    # Co-first authors: Liu Yajing, Zhang Shanshan, Li Chen contributed equally to this work.
Published:December 28, 2022DOI:https://doi.org/10.1016/j.trsl.2022.12.007

      Abstract

      Polycystic ovary syndrome (PCOS), characterized by the androgen excess and arrest of antral follicles, is a common endocrine disorder among women lacking specific diagnostic biomarkers and therapeutic targets. Herein, we studied the molecular mechanism of miR-96-5p in the process of PCOS and its potential applications in PCOS. Clinically, we found that miR-96-5p significantly decreased in serum, follicular fluid and primary human granulosa cells (hGCs) of PCOS patients (n = 70) vs non-PCOS women (n = 60), as well as in the ovaries of 3-types of induced PCOS-like mice. Furthermore, we demonstrated that the elevated circulating miR-96-5p levels were significantly correlated with the PCOS disordered endocrine clinical features, and the area under the curve of receiver operating characteristic was 0.8344, with 75.71% specificity and 80% sensitivity. Mechanically, we identified miR-96-5p as an androgen-regulated miRNA that directly targets the forkhead transcription factor FOXO1. Inhibition of miR-96-5p decreased estrogen synthesis, while decreasing the cell proliferation index of KGN via regulating the expression of FOXO1 and its downstream genes. Inversely, inhibition of FOXO1 abrogated the effect of miR-96-5p on estrogen synthesis and proliferation index. Of note, ovarian intra-bursal injection of miR-96-5p agomir rescued the phenotypes of dehydroepiandrosterone-induced PCOS like mice. In conclusion, our results clarified a vital role of miR-96-5p in the pathogenesis of PCOS and might serve as a novel diagnostic biomarker and therapeutic target for PCOS.

      Keywords

      Abbreviations:

      AMH (anti-Mullerian hormone), AR (androgen receptor), AREs (androgen response elements), AUC (area under the curve), BMI (body mass index), CMC (carboxymethylcellulose), CI (confidence interval), DHEA (dehydroepiandrosterone), DHT (dihydrotestosterone), E2 (estradiol), FF (follicular fluid), FSH (follicle-stimulating hormone), FT3 (free triiodothyronine), FT4 (free thyroxine), hGCs (human granulosa cells), INS (insulin), IVF (in-vitro fertilization), LH (luteinizing hormone), mGCs (mouse granulosa cells), miRNAs (microRNAs), MMP (mitochondrial membrane potential), NC (negative control), P (progesterone), PCOS (polycystic ovary syndrome), PI (propidium iodide), PNA (prenatal androgen treatment), PRL (prolactin), qPCR (quantitative real-time PCR), RBP (RNA-binding protein), ROC (receiver operating characteristic), T (testosterone), TSH (thyroid stimulating hormone), TFs (transcription factors), UTR (untranslated region)
      To read this article in full you will need to make a payment

      Purchase one-time access:

      Academic & Personal: 24 hour online accessCorporate R&D Professionals: 24 hour online access
      One-time access price info
      • For academic or personal research use, select 'Academic and Personal'
      • For corporate R&D use, select 'Corporate R&D Professionals'

      Subscribe:

      Subscribe to Translational Research
      Already a print subscriber? Claim online access
      Already an online subscriber? Sign in
      Institutional Access: Sign in to ScienceDirect

      References

        • Escobar-Morreale HF.
        Polycystic ovary syndrome: definition, aetiology, diagnosis and treatment.
        Natr rev Endocrinol. 2018; 14: 270-284
        • Azziz R.
        PCOS in 2015: new insights into the genetics of polycystic ovary syndrome.
        Natr rev Endocrinol. 2016; 12: 74-75
        • Comim FV
        • Teerds K
        • Hardy K
        • Franks S.
        Increased protein expression of LHCG receptor and 17α-hydroxylase/17-20-lyase in human polycystic ovaries.
        Human reprod (Oxford, England). 2013; 28: 3086-3092
        • Goodarzi MO
        • Dumesic DA
        • Chazenbalk G
        • Azziz R.
        Polycystic ovary syndrome: etiology, pathogenesis and diagnosis.
        Natr rev Endocrinol. 2011; 7: 219-231
        • Bertoldo MJ
        • Caldwell ASL
        • Riepsamen AH
        • et al.
        A hyperandrogenic environment causes intrinsic defects that are detrimental to follicular dynamics in a PCOS mouse Model.
        Endocrinology. 2019; 160: 699-715
        • Harlow CR
        • Shaw HJ
        • Hillier SG
        • Hodges JK.
        Factors influencing follicle-stimulating hormone-responsive steroidogenesis in marmoset granulosa cells: effects of androgens and the stage of follicular maturity.
        Endocrinology. 1988; 122: 2780-2787
        • Rachoń D.
        Differential diagnosis of hyperandrogenism in women with polycystic ovary syndrome.
        Exp and clin endocrinol & diabetes: offi j, German Soc Endocrinol [and] German Diabetes Assoc. 2012; 120: 205-209
        • Sander VA
        • Hapon MB
        • Sícaro L
        • Lombardi EP
        • Jahn GA
        • Motta AB.
        Alterations of folliculogenesis in women with polycystic ovary syndrome.
        The J steroid biochem and mol biol. 2011; 124: 58-64
        • Azziz R
        • Woods KS
        • Reyna R
        • Key TJ
        • Knochenhauer ES
        • Yildiz BO.
        The prevalence and features of the polycystic ovary syndrome in an unselected population.
        J Clin Endocrinol Metab. 2004; 89: 2745-2749
        • He L
        • Hannon GJ.
        MicroRNAs: small RNAs with a big role in gene regulation.
        Natr rev Genetics. 2004; 5: 522-531
        • Hayes J
        • Peruzzi PP
        • Lawler S.
        MicroRNAs in cancer: biomarkers, functions and therapy.
        Trends in mol med. 2014; 20: 460-469
        • Kalayinia S
        • Arjmand F
        • Maleki M
        • Malakootian M
        • Singh CP.
        MicroRNAs: roles in cardiovascular development and disease.
        Cardiovasc pathol: the off j Soc Cardiovasc Pathol. 2021; 50107296
        • Scheideler M.
        MicroRNAs in adipocyte formation and obesity.
        Best pract & res Clin endocrinol & metabol. 2016; 30: 653-664
        • Zhang S
        • Liu Y
        • Wang M
        • et al.
        Role and mechanism of miR-335-5p in the pathogenesis and treatment of polycystic ovary syndrome.
        Translational Res. 2022;
        • Wang M
        • Liu M
        • Sun J
        • et al.
        MicroRNA-27a-3p affects estradiol and androgen imbalance by targeting Creb1 in the granulosa cells in mouse polycytic ovary syndrome model.
        Reprod Biol. 2017; 17: 295-304
        • Wang M
        • Sun J
        • Xu B
        • et al.
        Functional characterization of MicroRNA-27a-3p expression in human polycystic ovary syndrome.
        Endocrinology. 2018; 159: 297-309
        • Hossain MM
        • Cao M
        • Wang Q
        • et al.
        Altered expression of miRNAs in a dihydrotestosterone-induced rat PCOS model.
        J ovarian res. 2013; 6: 36
        • Jiang X
        • Li J
        • Zhang B
        • Hu J
        • Ma J
        • Cui L
        • et al.
        Differential expression profile of plasma exosomal microRNAs in women with polycystic ovary syndrome.
        Fertility and sterility. 2021; 115: 782-792
        • Butler AE
        • Ramachandran V
        • Cunningham TK
        • David R
        • Gooderham NJ
        • Benurwar M
        • et al.
        Increased microRNA levels in women with polycystic ovarian syndrome but without insulin resistance: a pilot prospective study.
        Front endocrinol. 2020; 11571357
        • Scalici E
        • Traver S
        • Mullet T
        • et al.
        Circulating microRNAs in follicular fluid, powerful tools to explore in vitro fertilization process.
        Sci rep. 2016; 6: 24976
        • Zhen J
        • Li J
        • Li X
        • Wang X
        • Xiao Y
        • Sun Z
        • et al.
        Downregulating lncRNA NEAT1 induces proliferation and represses apoptosis of ovarian granulosa cells in polycystic ovary syndrome via microRNA-381/IGF1 axis.
        J biomed sci. 2021; 28: 53
        • Liu Y
        • Du SY
        • Ding M
        • et al.
        The BMP4-Smad signaling pathway regulates hyperandrogenism development in a female mouse model.
        The J biol chem. 2017; 292: 11740-11750
        • Kafali H
        • Iriadam M
        • Ozardali I
        • Demir N.
        Letrozole-induced polycystic ovaries in the rat: a new model for cystic ovarian disease.
        Arch med res. 2004; 35: 103-108
        • Sullivan SD
        • Moenter SM.
        Prenatal androgens alter GABAergic drive to gonadotropin-releasing hormone neurons: implications for a common fertility disorder.
        Proc Natl Acad Sci United States of Am. 2004; 101: 7129-7134
        • Liu G
        • Liu S
        • Xing G
        • Wang F.
        lncRNA PVT1/MicroRNA-17-5p/PTEN axis regulates secretion of E2 and P4, proliferation, and apoptosis of ovarian granulosa cells in PCOS.
        Mol therapy Nucleic acids. 2020; 20: 205-216
        • Anagnostis P
        • Tarlatzis BC
        • Kauffman RP.
        Polycystic ovarian syndrome (PCOS): Long-term metabolic consequences.
        Metabol: clin and exp. 2018; 86: 33-43
        • Markström E
        • Svensson E
        • Shao R
        • Svanberg B
        • Billig H.
        Survival factors regulating ovarian apoptosis – dependence on follicle differentiation.
        Reprod (Cambridge, England). 2002; 123: 23-30
        • Gong Y
        • Luo S
        • Fan P
        • Zhu H
        • Li Y
        • Huang W.
        Growth hormone activates PI3K/Akt signaling and inhibits ROS accumulation and apoptosis in granulosa cells of patients with polycystic ovary syndrome.
        Reprod biol and endocrinol: RB&E. 2020; 18: 121
        • Wilhelm K
        • Happel K
        • Eelen G
        • et al.
        FOXO1 couples metabolic activity and growth state in the vascular endothelium.
        Nature. 2016; 529: 216-220
        • Fu Z
        • Tindall DJ.
        FOXOs, cancer and regulation of apoptosis.
        Oncogene. 2008; 27: 2312-2319
        • Xu R
        • Wang Z.
        Involvement of transcription factor FoxO1 in the pathogenesis of polycystic ovary syndrome.
        Front physiol. 2021; 12649295
        • Huang J
        • Zhao J
        • Geng X
        • et al.
        Long non-coding RNA lnc-CCNL1-3:1 promotes granulosa cell apoptosis and suppresses glucose uptake in women with polycystic ovary syndrome.
        Mol therapy Nucleic acids. 2021; 23: 614-628
        • Sørensen AE
        • Wissing ML
        • Salö S
        • Englund AL
        • Dalgaard LT.
        MicroRNAs related to polycystic ovary syndrome (PCOS).
        Genes. 2014; 5: 684-708
      1. Tamaddon M, Azimzadeh M and Tavangar SM., microRNAs and long non-coding RNAs as biomarkers for polycystic ovary syndrome. J cellular and mol med 2022;26:654–70.

        • Leung WK
        • He M
        • Chan AW
        • Law PT
        • Wong N.
        Wnt/β-Catenin activates MiR-183/96/182 expression in hepatocellular carcinoma that promotes cell invasion.
        Cancer letters. 2015; 362: 97-105
        • Long MD
        • Singh PK
        • Russell JR
        • Llimos G
        • Rosario S
        • Rizvi A
        • et al.
        The miR-96 and RARγ signaling axis governs androgen signaling and prostate cancer progression.
        Oncogene. 2019; 38: 421-444
        • Zhou HY
        • Wu CQ
        • Bi EX.
        MiR-96-5p inhibition induces cell apoptosis in gastric adenocarcinoma.
        World j gastroenterol. 2019; 25: 6823-6834
        • Luo X
        • He X
        • Liu X
        • Zhong L
        • Hu W.
        miR-96-5p suppresses the progression of nasopharyngeal carcinoma by targeting CDK1.
        OncoTargets and therap. 2020; 13: 7467-7477
        • Ding L
        • Gao F
        • Zhang M
        • et al.
        Higher PDCD4 expression is associated with obesity, insulin resistance, lipid metabolism disorders, and granulosa cell apoptosis in polycystic ovary syndrome.
        Fertility and sterility. 2016; 105 (e3): 1330-1337
        • Zhang X
        • Xiao H
        • Zhang X
        • et al.
        Decreased microRNA-125b-5p disrupts follicle steroidogenesis through targeting PAK3/ERK1/2 signalling in mouse preantral follicles.
        Metabol: clin and exp. 2020; 107154241
        • Liu Z
        • Rudd MD
        • Hernandez-Gonzalez I
        • et al.
        FSH and FOXO1 regulate genes in the sterol/steroid and lipid biosynthetic pathways in granulosa cells.
        Mol endocrinol (Baltimore, Md). 2009; 23: 649-661
        • Kinyua AW
        • Doan KV
        • Yang DJ
        • et al.
        Insulin regulates adrenal steroidogenesis by stabilizing SF-1 activity.
        Sci rep. 2018; 8: 5025