Extracellular vesicles from iPSC-MSCs alleviate chemotherapy-induced mouse ovarian damage via the ILK-PI3K/AKT pathway

Chemotherapy can significantly reduce follicle counts in ovarian tissues and damage ovarian stroma, causing endocrine disorder, reproductive dysfunction, and primary ovarian insufficiency (POI). Recent studies have suggested that extracellular vesicles (EVs) secreted from mesenchymal stem cells (MSCs) exert therapeutic effects in various degenerative diseases. In this study, transplantation of EVs from human induced pluripotent stem cell-derived MSCs (iPSC-MSC-EVs) resulted in significant restoration of ovarian follicle numbers, improved granulosa cell proliferation, and inhibition of apoptosis in chemotherapy-damaged granulosa cells, cultured ovaries, and in vivo ovaries in mice. Mechanistically, treatment with iPSC-MSC-EVs resulted in up-regulation of the integrin-linked kinase (ILK) -PI3K/AKT pathway, which is suppressed during chemotherapy, most likely through the transfer of regulatory microRNAs (miRNAs) targeting ILK pathway genes. This work provides a framework for the development of advanced therapeutics to ameliorate ovarian damage and POI in female chemotherapy patients.

[1]  A. Mazur,et al.  Integrin-linked kinase (ILK): the known vs. the unknown and perspectives , 2022, Cellular and Molecular Life Sciences.

[2]  Haijie Han,et al.  Exosomes-loaded thermosensitive hydrogels for corneal epithelium and stroma regeneration. , 2021, Biomaterials.

[3]  M. Stevens,et al.  Extracellular vesicles for tissue repair and regeneration: evidence, challenges and opportunities. , 2021, Advanced drug delivery reviews.

[4]  Abdelrahman Yousif,et al.  Premature ovarian insufficiency: pathogenesis and therapeutic potential of mesenchymal stem cell , 2021, Journal of Molecular Medicine.

[5]  Yan Zhou,et al.  Advances in mesenchymal stem cell exosomes: a review , 2021, Stem cell research & therapy.

[6]  Qingwei Zhu,et al.  Small extracellular vesicles secreted by human iPSC-derived MSC enhance angiogenesis through inhibiting STAT3-dependent autophagy in ischemic stroke , 2020, Stem Cell Research & Therapy.

[7]  Jiafeng Lu,et al.  Exosomal miRNA‐17‐5p derived from human umbilical cord mesenchymal stem cells improves ovarian function in premature ovarian insufficiency by regulating SIRT7 , 2020, Stem cells.

[8]  Jiafeng Lu,et al.  Exosomal miRNA-320a Is Released from hAMSCs and Regulates SIRT4 to Prevent Reactive Oxygen Species Generation in POI , 2020, Molecular therapy. Nucleic acids.

[9]  N. Arsenijević,et al.  Mesenchymal Stem Cell-Derived Exosomes and Other Extracellular Vesicles as New Remedies in the Therapy of Inflammatory Diseases , 2019, Cells.

[10]  Ying-pu Sun,et al.  miR-644-5p carried by bone mesenchymal stem cell-derived exosomes targets regulation of p53 to inhibit ovarian granulosa cell apoptosis , 2019, Stem Cell Research & Therapy.

[11]  N. Binart,et al.  The Impact of Chemotherapy on the Ovaries: Molecular Aspects and the Prevention of Ovarian Damage , 2019, International journal of molecular sciences.

[12]  Yueqin Liu,et al.  Bone marrow mesenchymal stem cell-derived exosomal miR-144-5p improves rat ovarian function after chemotherapy-induced ovarian failure by targeting PTEN , 2019, Laboratory Investigation.

[13]  Xin Du,et al.  Therapeutic effects of human umbilical cord mesenchymal stem cell-derived microvesicles on premature ovarian insufficiency in mice , 2019, Stem Cell Research & Therapy.

[14]  Shixia Bu,et al.  Human Amniotic Epithelial Cell-Derived Exosomes Restore Ovarian Function by Transferring MicroRNAs against Apoptosis , 2019, Molecular therapy. Nucleic acids.

[15]  W. Shen,et al.  Cytotoxicity and DNA Damage Caused from Diazinon Exposure by Inhibiting the PI3K-AKT Pathway in Porcine Ovarian Granulosa Cells. , 2018, Journal of agricultural and food chemistry.

[16]  Zi-jiang Chen,et al.  Molecular Genetics of Premature Ovarian Insufficiency , 2018, Trends in Endocrinology & Metabolism.

[17]  Jiafeng Lu,et al.  Exosomes derived from human adipose mesenchymal stem cells improve ovary function of premature ovarian insufficiency by targeting SMAD , 2018, Stem Cell Research & Therapy.

[18]  Jenq-Wen Huang,et al.  The Small Molecule Inhibitor QLT-0267 Decreases the Production of Fibrin-Induced Inflammatory Cytokines and Prevents Post-Surgical Peritoneal Adhesions , 2018, Scientific Reports.

[19]  Xiaomin He,et al.  Exosomes and regenerative medicine: state of the art and perspectives. , 2018, Translational research : the journal of laboratory and clinical medicine.

[20]  L. Dagnino,et al.  Protective role of integrin-linked kinase against oxidative stress and in maintenance of genomic integrity , 2018, Oncotarget.

[21]  Shixia Bu,et al.  Paracrine effects of human amniotic epithelial cells protect against chemotherapy-induced ovarian damage , 2017, Stem Cell Research & Therapy.

[22]  Dali Li,et al.  Mitochondrial Function Regulated by Mitoguardin-1/2 Is Crucial for Ovarian Endocrine Functions and Ovulation , 2017, Endocrinology.

[23]  B. Kong,et al.  Exosomes derived from human umbilical cord mesenchymal stem cells protect against cisplatin-induced ovarian granulosa cell stress and apoptosis in vitro , 2017, Scientific Reports.

[24]  G. Camussi,et al.  The effects of glomerular and tubular renal progenitors and derived extracellular vesicles on recovery from acute kidney injury , 2017, Stem Cell Research & Therapy.

[25]  K. Sugiura,et al.  Oocyte-dependent activation of MTOR in cumulus cells controls the development and survival of cumulus–oocyte complexes , 2016, Journal of Cell Science.

[26]  X. Niu,et al.  Exosomes Secreted by Human-Induced Pluripotent Stem Cell-Derived Mesenchymal Stem Cells Repair Critical-Sized Bone Defects through Enhanced Angiogenesis and Osteogenesis in Osteoporotic Rats , 2016, International journal of biological sciences.

[27]  A. Pozzi,et al.  Integrin-Linked Kinase in Muscle Is Necessary for the Development of Insulin Resistance in Diet-Induced Obese Mice , 2016, Diabetes.

[28]  Joydeep Basu,et al.  Exosomes for repair, regeneration and rejuvenation , 2016, Expert opinion on biological therapy.

[29]  Zi-jiang Chen,et al.  Genetics of primary ovarian insufficiency: new developments and opportunities , 2015, Human reproduction update.

[30]  Artemis G. Hatzigeorgiou,et al.  DIANA-miRPath v3.0: deciphering microRNA function with experimental support , 2015, Nucleic Acids Res..

[31]  D. Dixon,et al.  Tamoxifen Prevents Apoptosis and Follicle Loss from Cyclophosphamide in Cultured Rat Ovaries1 , 2015, Biology of reproduction.

[32]  Changqing Zhang,et al.  Exosomes secreted by human-induced pluripotent stem cell-derived mesenchymal stem cells attenuate limb ischemia by promoting angiogenesis in mice , 2015, Stem Cell Research & Therapy.

[33]  Changqing Zhang,et al.  Exosomes released from human induced pluripotent stem cells-derived MSCs facilitate cutaneous wound healing by promoting collagen synthesis and angiogenesis , 2015, Journal of Translational Medicine.

[34]  A. Möller,et al.  Optimized exosome isolation protocol for cell culture supernatant and human plasma , 2015, Journal of extracellular vesicles.

[35]  Andreas Krämer,et al.  Causal analysis approaches in Ingenuity Pathway Analysis , 2013, Bioinform..

[36]  H. Kanety,et al.  Cyclophosphamide Triggers Follicle Activation and “Burnout”; AS101 Prevents Follicle Loss and Preserves Fertility , 2013, Science Translational Medicine.

[37]  S. Yamanaka,et al.  An Efficient Nonviral Method to Generate Integration‐Free Human‐Induced Pluripotent Stem Cells from Cord Blood and Peripheral Blood Cells , 2013, Stem cells.

[38]  C. Yao,et al.  Targeting integrin-linked kinase increases apoptosis and decreases invasion of myeloma cell lines and inhibits IL-6 and VEGF secretion from BMSCs , 2011, Medical oncology.

[39]  Yasuko Matsumura,et al.  A more efficient method to generate integration-free human iPS cells , 2011, Nature Methods.

[40]  W. Wallace,et al.  Toxicity of Chemotherapy and Radiation on Female Reproduction , 2010, Clinical obstetrics and gynecology.

[41]  A. Shelling.  Premature ovarian failure. , 2010, Reproduction.

[42]  S. Hehlgans,et al.  The Small Molecule Inhibitor QLT0267 Radiosensitizes Squamous Cell Carcinoma Cells of the Head and Neck , 2009, PloS one.

[43]  Thomas Helleday,et al.  DNA repair pathways as targets for cancer therapy , 2008, Nature Reviews Cancer.

[44]  C. J. Barnes,et al.  Phosphorylation-dependent regulation of nuclear localization and functions of integrin-linked kinase , 2007, Proceedings of the National Academy of Sciences.

[45]  F. Broekmans,et al.  Female reproductive ageing: current knowledge and future trends , 2007, Trends in Endocrinology & Metabolism.

[46]  S. Hehlgans,et al.  An essential role of integrin-linked kinase in the cellular radiosensitivity of normal fibroblasts during the process of cell adhesion and spreading , 2007, International journal of radiation biology.

[47]  P. Devine,et al.  Characterizing the ovotoxicity of cyclophosphamide metabolites on cultured mouse ovaries. , 2006, Toxicological sciences : an official journal of the Society of Toxicology.

[48]  J. Rowe,et al.  Fertility and ovarian function are preserved in women treated with an intensified regimen of cyclophosphamide, adriamycin, vincristine and prednisone (Mega-CHOP) for non-Hodgkin lymphoma. , 2005, Human reproduction.

[49]  S. Dedhar,et al.  Integration of cell attachment, cytoskeletal localization, and signaling by integrin-linked kinase (ILK), CH-ILKBP, and the tumor suppressor PTEN. , 2003, Molecular biology of the cell.

[50]  S. Dedhar,et al.  Inhibition of integrin-linked kinase (ILK) suppresses activation of protein kinase B/Akt and induces cell cycle arrest and apoptosis of PTEN-mutant prostate cancer cells. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[51]  S. Dedhar,et al.  Overexpression of the Integrin-linked Kinase Promotes Anchorage-independent Cell Cycle Progression* , 1997, The Journal of Biological Chemistry.

[52]  W. Remmele,et al.  [Recommendation for uniform definition of an immunoreactive score (IRS) for immunohistochemical estrogen receptor detection (ER-ICA) in breast cancer tissue]. , 1987, Der Pathologe.

[53]  J. Annegers,et al.  Incidence of Premature Ovarian Failure , 1986, Obstetrics and gynecology.

[54]  T. Pedersen,et al.  Proposal for a classification of oocytes and follicles in the mouse ovary. , 1968, Journal of reproduction and fertility.

[55]  Wei-hong Zhang,et al.  [Granulosa cells model transfected by ENO1 overexpression adenovirus vector and the effect of ENO1 overexpression on progesterone secretion]. , 2014, Zhongguo ying yong sheng li xue za zhi = Zhongguo yingyong shenglixue zazhi = Chinese journal of applied physiology.

[56]  X. Fu,et al.  Bone marrow mesenchymal stem cell transplantation improves ovarian function and structure in rats with chemotherapy-induced ovarian damage. , 2008, Cytotherapy.

[57]  G. Mills,et al.  Effects of the integrin-linked kinase inhibitor QLT0267 on squamous cell carcinoma of the head and neck. , 2007, Archives of otolaryngology--head & neck surgery.

[58]  D. Prockop,et al.  Minimal criteria for defining multipotent mesenchymal stromal cells. The International Society for Cellular Therapy position statement. , 2006, Cytotherapy.

[59]  R. Setlow.  Dna repair pathways. , 1980, Basic life sciences.

[60]  Thomas D. Schmittgen,et al.  Analysis of Relative Gene Expression Data Using Real-Time Quantitative PCR and the 2 2 DD C T Method , 2022 .