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Role of the PI3K and Hippo pathways in follicle activation after grafting of human ovarian tissue

  • Reproductive Physiology and Disease
  • Published:
Journal of Assisted Reproduction and Genetics Aims and scope Submit manuscript

Abstract

Purpose

Our aim was to elucidate the mechanisms involved in follicle activation of the ovarian reserve after human ovarian tissue transplantation, with specific focus on the role of the effectors of the PI3K (mTOR and FOXO1) and Hippo (YAP) signaling pathways and whether they are somehow altered.

Methods

Frozen-thawed ovarian tissue was collected from six women (age 25–35 years) undergoing surgery for non-ovarian pathologies and divided into 4 fragments in each case: one for non-grafted controls and three for grafting to immunodeficient mice for 3, 7 and 21 days. The tissue was processed for hematoxylin and eosin staining, immunohistochemistry and immunofluorescence at different timepoints before and after grafting. Activation of the PI3K and Hippo signaling pathways was investigated by analysis of mTOR phosphorylation, FOXO1 cytoplasmic localization and YAP nuclear localization.

Results

No change in mTOR levels was observed in primordial follicles post-transplantation, but a significant upturn was recorded in growing follicles compared with primordial follicles, irrespective of grafting time. A higher percentage of primordial follicles was also found with FOXO1 in the cytoplasm after 3 days of transplantation than in non-grafted controls. Finally, a greater proportion of primordial follicles was detected with YAP in the nucleus at all timepoints after grafting.

Conclusions

This study supports the hypothesis that follicle activation may occur as an early event after transplantation, with follicle growth and death both contributing to the burnout phenomenon. This is the first time that the effectors of the PI3K and Hippo pathways have been investigated in grafted human ovarian tissue and their role in burnout documented.

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References

  1. Wallace WHB, Anderson RA, Irvine DS. Fertility preservation for young patients with cancer: who is at risk and what can be offered? Lancet Oncol. 2005;6:209–18.

    Article  Google Scholar 

  2. Donnez J, Dolmans MM. Fertility preservation in women. N Engl J Med. 2017;377:1657–65.

    Article  Google Scholar 

  3. Manavella DD, Cacciottola L, Desmet CM, Jordan BF, Donnez J, Amorim CA, et al. Adipose tissue-derived stem cells in a fibrin implant enhance neovascularization in a peritoneal grafting site: a potential way to improve ovarian tissue transplantation. Hum Reprod. 2018;33:270–9.

    Article  CAS  Google Scholar 

  4. Manavella DD, Cacciottola L, Pomme S, Desmet CM, Jordan BF, Donnez J, et al. Two-step transplantation with adipose tissue-derived stem cells increases follicle survival by enhancing vascularization in xenografted frozen-thawed human ovarian tissue. Hum Reprod. 2018;33:1107–16.

    Article  CAS  Google Scholar 

  5. Dolmans MM, Martinez-Madrid B, Gadisseux E, Guiot Y, Yuan WY, Torre A, et al. Short-term transplantation of isolated human ovarian follicles and cortical tissue into nude mice. Reproduction. 2007;134:253–62.

    Article  CAS  Google Scholar 

  6. Gavish Z, Spector I, Peer G, Schlatt S, Wistuba J, Roness H, et al. Follicle activation is a significant and immediate cause of follicle loss after ovarian tissue transplantation. J Assist Reprod Genet. 2018;35:61–9.

    Article  Google Scholar 

  7. Masciangelo R, Hossay C, Donnez J, Dolmans MM. Does the Akt pathway play a role in follicle activation after grafting of human ovarian tissue? Reprod BioMed Online. 2019;39(2):196–8.

    Article  CAS  Google Scholar 

  8. Song G, Ouyang G, Bao S. The activation of Akt/PKB signaling pathway and cell survival. J Cell Mol Med. 2005;9:59–71.

    Article  CAS  Google Scholar 

  9. Castrillon DH, Miao L, Kollipara R, Horner JW, DePinho RA. Suppression of ovarian follicle activation in mice by the transcription factor Foxo3a. Science. 2003;301:215–8.

    Article  CAS  Google Scholar 

  10. John GB, Gallardo TD, Shirley LJ, Castrillon DH. Foxo3 is a PI3K-dependent molecular switch controlling the initiation of oocyte growth. Dev Biol. 2008;321:197–204.

    Article  CAS  Google Scholar 

  11. Reddy P, Liu L, Adhikari D, Jagarlamudi K, Rajareddy S, Shen Y, et al. Oocyte-specific deletion of Pten causes premature activation of the primordial follicle pool. Science. 2008;319:611–3.

    Article  CAS  Google Scholar 

  12. Adhikari D, Flohr G, Gorre N, Shen Y, Yang H, Lundin E, et al. Disruption of Tsc2 in oocytes leads to overactivation of the entire pool of primordial follicles. Mol Hum Reprod. 2009;15:765–70.

    Article  CAS  Google Scholar 

  13. Adhikari D, Zheng W, Shen Y, Gorre N, Hamalainen T, Cooney AJ, et al. Tsc/mTORC1 signaling in oocytes governs the quiescence and activation of primordial follicles. Hum Mol Genet. 2010;19:397–410.

    Article  CAS  Google Scholar 

  14. Adhikari D, Gorre N, Risal S, Zhao Z, Zhang H, Shen Y, et al. The safe use of a PTEN inhibitor for the activation of dormant mouse primordial follicles and generation of fertilizable eggs. PLoS One. 2012;7:e39034.

    Article  CAS  Google Scholar 

  15. Li J, Kawamura K, Cheng Y, Liu S, Klein C, Liu S, et al. Activation of dormant ovarian follicles to generate mature eggs. Proc Natl Acad Sci U S A. 2010;107:10280–4.

    Article  CAS  Google Scholar 

  16. Zhang H, Liu K. Cellular and molecular regulation of the activation of mammalian primordial follicles: somatic cells initiate follicle activation in adulthood. Hum Reprod Update. 2015;21:779–86.

    Article  CAS  Google Scholar 

  17. Hsueh AJ, Kawamura K, Cheng Y, Fauser BC. Intraovarian control of early folliculogenesis. Endocr Rev. 2015;36:1–24.

    Article  CAS  Google Scholar 

  18. Kawamura K, Cheng Y, Suzuki N, Deguchi M, Sato Y, Takae S, et al. Hippo signaling disruption and Akt stimulation of ovarian follicles for infertility treatment. Proc Natl Acad Sci U S A. 2013;110:17474–9.

    Article  CAS  Google Scholar 

  19. Gosden RG, Baird DT, Wade JC, Webb R. Restoration of fertility to oophorectomized sheep by ovarian autografts stored at -196 degrees C. Hum Reprod. 1994;9:597–603.

    Article  CAS  Google Scholar 

  20. Donnez J, Dolmans MM, Demylle D, Jadoul P, Pirard C, Squifflet J, et al. Livebirth after orthotopic transplantation of cryopreserved ovarian tissue. Lancet. 2004;364:1405–10.

    Article  CAS  Google Scholar 

  21. Amorim CA, Dolmans MM, David A, Jaeger J, Vanacker J, Camboni A, et al. Vitrification and xenografting of human ovarian tissue. Fertil Steril. 2012;98:1291–8.

    Article  Google Scholar 

  22. Hewitt SM, Baskin DG, Frevert CW, Stahl WL, Rosa-Molinar E. Controls for immunohistochemistry: the Histochemical Society’s standards of practice for validation of immunohistochemical assays. J Histochem Cytochem. 2014;62:693–7.

    Article  CAS  Google Scholar 

  23. Bouzin C, Saini ML, Khaing KK, Ambroise J, Marbaix E, Gregoire V, et al. Digital pathology: elementary, rapid and reliable automated image analysis. Histopathology. 2016;68:888–96.

    Article  Google Scholar 

  24. Courtoy GE, Donnez J, Marbaix E, Barreira M, Luyckx M, Dolmans MM. Progesterone receptor isoforms, nuclear corepressor-1 and steroid receptor coactivator-1 and b-cell lymphoma 2 and Akt and Akt phosphorylation status in uterine myomas after ulipristal acetate treatment: a systematic immunohistochemical evaluation. Gynecol Obstet Invest. 2018;83:443–54.

    Article  Google Scholar 

  25. Van Eyck AS, Jordan BF, Gallez B, Heilier JF, Van Langendonckt A, Donnez J. Electron paramagnetic resonance as a tool to evaluate human ovarian tissue reoxygenation after xenografting. Fertil Steril. 2009;92:374–81.

    Article  Google Scholar 

  26. Van Eyck AS, Bouzin C, Feron O, Romeu L, Van Langendonckt A, Donnez J, et al. Both host and graft vessels contribute to revascularization of xenografted human ovarian tissue in a murine model. Fertil Steril. 2010;93:1676–85.

    Article  Google Scholar 

  27. Ting AY, Zelinski MB. Characterization of FOXO1, 3 and 4 trascription factors in ovaruies of fetal, prepubertal and adult rhesus macaques. Biol Reprod. 2017;96:1052–9.

    Article  Google Scholar 

  28. Laplante M, Sabatini DM. mTOR signaling. Cold Spring Harb Perspect Biol. 2012;4.

    Google Scholar 

  29. Saxton RA, Sabatini DM. mTOR signaling in growth, metabolism, and disease. Cell. 2017;168:960–76.

    Article  CAS  Google Scholar 

  30. Novella-Maestre E, Herraiz S, Rodriguez-Iglesias B, Diaz-Garcia C, Pellicer A. Short-term PTEN inhibition improves in vitro activation of primordial follicles, preserves follicular viability, and restores amh levels in cryopreserved ovarian tissue from cancer patients. PLoS One. 2015;10:e0127786.

    Article  Google Scholar 

  31. Suzuki N, Yoshioka N, Takae S, Sugishita Y, Tamura M, Hashimoto S, et al. Successful fertility preservation following ovarian tissue vitrification in patients with primary ovarian insufficiency. Hum Reprod. 2015;30:608–15.

    Article  Google Scholar 

  32. Zhai J, Yao G, Dong F, Bu Z, Cheng Y, Sato Y, et al. In vitro activation of follicles and fresh tissue auto-transplantation in primary ovarian insufficiency patients. J Clin Endocrinol Metab. 2016;101:4405–12.

    Article  CAS  Google Scholar 

  33. Dolmans MM, Cordier F, Amorim CA, Donnez J, Vander LC. In vitro activation prior to transplantation of human ovarian tissue: is it truly effective? Front Endocrinol (Lausanne). 2019;10:520.

    Article  Google Scholar 

  34. Pan D. Hippo signaling in organ size control. Genes Dev. 2007;21:886–97.

    Article  CAS  Google Scholar 

  35. Hergovich A. Mammalian Hippo signalling: a kinase network regulated by protein-protein interactions. Biochem Soc Trans. 2012;40:124–8.

    Article  CAS  Google Scholar 

  36. Holbourn KP, Acharya KR, Perbal B. The CCN family of proteins: structure-function relationships. Trends Biochem Sci. 2008;33:461–73.

    Article  CAS  Google Scholar 

  37. Liu-Chittenden Y, Huang B, Shim JS, Chen Q, Lee SJ, Anders RA, et al. Genetic and pharmacological disruption of the TEAD-YAP complex suppresses the oncogenic activity of YAP. Genes Dev. 2012;26:1300–5.

    Article  CAS  Google Scholar 

  38. Roness H, Gavish Z, Cohen Y, Meirow D. Ovarian follicle burnout: a universal phenomenon? Cell Cycle. 2013;12:3245–6.

    Article  CAS  Google Scholar 

  39. Guertin DA, Sabatini DM. Defining the role of mTOR in cancer. Cancer Cell. 2007;12(1):9–22.

    Article  CAS  Google Scholar 

Download references

Acknowledgments

The authors thank Mira Hryniuk, B.A., for reviewing the English language of the article, and Dolores Gonzalez and Olivier Van Kerk for their technical assistance.

Author contribution statement

R.M.: conception and design of the study, experimental procedures, analysis of results, statistical analysis and article preparation; C.H.: experimental procedures; M.C.C.: analysis of results and discussion contribution; D.D.M.: experimental procedures, analysis of results and discussion contribution; C.A.A.: discussion contribution; J.D.: data evaluation, discussion contribution and article revision; M.M.D.: conception of the study, data evaluation, discussion contribution and article revision.

Funding

This study was supported by grants from the Fonds National de la Recherche Scientifique de Belgique (FNRS-PDR Convention T.0077.14, Télévie grant 7.4590.16 awarded to Rossella Masciangelo, EOS grant 30443682 to Maria Costanza Chiti, Télévie grant no. 7.6515.16 F to Diego Daniel Manavella and 5/4/150/5 grant to Marie-Madeleine Dolmans; CAA is an FSR-FNRS research associate), the Fonds Speciaux de Recherche, and the Foundation Against Cancer.

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Authors and Affiliations

  1. Pôle de Recherche en Gynécologie, Institut de Recherche Expérimentale et Clinique, Université Catholique de Louvain, Avenue Mounier 52, bte. B1.52.02, 1200, Brussels, Belgium

    Rossella Masciangelo, Camille Hossay, Maria Costanza Chiti, Diego Daniel Manavella, Christiani Andrade Amorim & Marie-Madeleine Dolmans

  2. Société de Recherche pour l’Infertilité, Avenue Grandchamp 143, 1150, Brussels, Belgium

    Jacques Donnez

  3. Département de Gynécologie, Cliniques Universitaires St. Luc, Avenue Hippocrate 10, 1200, Brussels, Belgium

    Marie-Madeleine Dolmans

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  1. Rossella Masciangelo
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  2. Camille Hossay
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  3. Maria Costanza Chiti
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  4. Diego Daniel Manavella
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  6. Jacques Donnez
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  7. Marie-Madeleine Dolmans
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Correspondence to Marie-Madeleine Dolmans.

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Masciangelo, R., Hossay, C., Chiti, M.C. et al. Role of the PI3K and Hippo pathways in follicle activation after grafting of human ovarian tissue. J Assist Reprod Genet 37, 101–108 (2020). https://doi.org/10.1007/s10815-019-01628-1

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  • DOI: https://doi.org/10.1007/s10815-019-01628-1

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