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Reproductive Sciences

, Volume 19, Issue 2, pp 143–151 | Cite as

Cross Talk Between Estradiol and mTOR Kinase in the Regulation of Ovarian Granulosa Proliferation

  • James Yu
  • Travis C. Thomson
  • Joshua JohnsonEmail author
Original Articles

Abstract

Treatment of ovarian granulosa cells and follicles with the mammalian target of rapamycin (mTOR) kinase inhibitor results in biphasic effects where nanomolar rapamycin (RAP) results in reduced proliferation, mitotic anomalies, and attenuated follicle growth, while the picomolar RAP results in accelerated follicle growth. Here, we tested whether such effects are specific to RAP or could be mimicked by 2 alternative mTOR inhibitors, everolimus (EV) and temsirolimus (TEM), and whether these effects were dependent on the presence of estradiol (E2). Spontaneously immortalized rat granulosa cells (SIGCs) were cultured in dose curves of RAP, EV, TEM, or vehicle with or without E2. Proliferation and phosphorylation of mTOR targets p70S6 kinase and 4E-binding protein (BP) were determined. Cell cycle gene array analysis and confirmatory quantitative reverse transcriptase polymerase chain reaction were performed upon cells treated with picomolar RAP versus controls. Nanomolar RAP, EV, and TEM reduced SIGC proliferation and decreased phospho-p70 and 4E-BP. Picomolar concentrations accelerated proliferation without affecting mTOR substrate phosphorylation. Acceleration of growth by picomolar inhibitor required E2. Picomolar drug treatment altered the transcription of cell cycle regulators, increasing Integrin beta 1 and calcineurin expression, and decreasing inhibin alpha, Chek1, p16ARF, p27/Kip1, and Sestrin2 expression. At nanomolar concentrations, mTOR inhibitors attenuated granulosa proliferation. Accelerated growth and alterations in cell cycle gene transcription found with picomolar concentrations required E2 within the intrafollicular concentration range. The low concentrations of inhibitors required to increase granulosa proliferation suggest a novel use to support the growth of ovarian follicles.

Keywords

ovary follicle growth folliculogenesis mTOR estradiol 

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References

  1. 1.
    Jacinto E, Loewith R, Schmidt A, et al. Mammalian TOR complex 2 controls the actin cytoskeleton and is rapamycin insensitive. Nat Cell Biol. 2004;6(11):1122–1128.Google Scholar
  2. 2.
    Wiederrecht GJ, Sabers CJ, Brunn GJ, Martin MM, Dumont FJ, Abraham RT. Mechanism of action of rapamycin: new insights into the regulation of G1-phase progression in eukaryotic cells. Prog Cell Cycle Res. 1995;1:53–71.Google Scholar
  3. 3.
    Wullschleger S, Loewith R, Hall MN. Tor signaling in growth and metabolism. Cell. 2006;124(3):471–484.Google Scholar
  4. 4.
    Kirchner G, Meier-Wiedenbach I, Manns M. Clinical pharmacokinetics of everolimus. Clin Pharmacokinet. 2004;43(2): 83–95.CrossRefGoogle Scholar
  5. 5.
    Pangas SA. Growth factors in ovarian development. Semin Reprod Med. 2007;25(4):225–234.Google Scholar
  6. 6.
    Sato E, Kimura N, Yokoo M, Miyake Y, Ikeda JE. Morphodynamics of ovarian follicles during oogenesis in mice. Microsc Res Tech. 2006;69(6):427–435.Google Scholar
  7. 7.
    Alam H, Maizels E, Park Y, et al. Follicle-stimulating hormone activation of hypoxia-inducible factor-1 by the phosphatidylinositol 3-kinase/AKT/Ras homolog enriched in brain (Rheb)/mammalian target of rapamycin (mTOR) pathway is necessary for induction of select protein markers of follicular differentiation. J Biol Chem 2004;279:19431–19440.Google Scholar
  8. 8.
    Kayampilly PP, Menon KM. Follicle-stimulating hormone increases tuberin phosphorylation and mammalian target of rapamycin signaling through an extracellular signal-regulated kinase-dependent pathway in rat granulosa cells. Endocrinology 2007;148:3950–3957.Google Scholar
  9. 9.
    Ponticelli C. The pleiotropic effects of mTor inhibitors. J Nephrol. 2004;17(6):762–768.Google Scholar
  10. 10.
    Song J, Salek-Ardakani S, So T, Croft M. The kinases aurora B and mTOR regulate the G1-S cell cycle progression of T lymphocytes. Nat Immunol. 2007;8(1):64–73.Google Scholar
  11. 11.
    Yu J, Yaba A, Kasiman C, Thomson T, Johnson J. mTOR controls ovarian follicle growth by regulating granulosa cell proliferation. PLoS ONE. 2011;6, 7:e21415.Google Scholar
  12. 12.
    Yaba A, Bianchi V, Borini A, Johnson J. A putative mitotic checkpoint dependent on mTOR function controls cell proliferation and survival in ovarian granulosa cells. Reprod Sci. 2008; 15(2):128–138.Google Scholar
  13. 13.
    Stewnius Y, Gorunova L, Jonson T, et al. Structural and numerical chromosome changes in colon cancer develop through telomere-mediated anaphase bridges, not through mitotic multipolarity. Proc Natl Acad Sci U S A. 2005;102(15):5541–5546.Google Scholar
  14. 14.
    Stein L, Stoica G, Tilley R, Burghardt R. Rat ovarian granulosa cell culture: a model system for the study of cell-cell communication during multistep transformation. Cancer Res. 1991;51(2): 696–706.Google Scholar
  15. 15.
    Akcakanat A, Singh G, Hung M, Meric-Bernstam F. Rapamycin regulates the phosphorylation of rictor. Biochem Biophys Res Commun. 2007;362(2):330–333.Google Scholar
  16. 16.
    Barilli A, Visigalli R, Sala R, et al. In human endothelial cells rapamycin causes mTORC2 inhibition and impairs cell viability and function. Cardiovasc Res. 2008;78(3):563–571.Google Scholar
  17. 17.
    Sarbassov D, Ali S, Sengupta S, et al. Prolonged rapamycin treatment inhibits mTORC2 assembly and Akt/PKB. Mol Cell. 2006; 22(2):159–168.Google Scholar
  18. 18.
    Gingras AC, Raught B, Sonenberg N. Regulation of translation initiation by FRAP/mTOR. Genes Dev. 2001;15(7):807–826.Google Scholar
  19. 19.
    Dmitriev S, Terenin I, Dunaevsky Y, Merrick W, Shatsky I. Assembly of 48S translation initiation complexes from purified components with mRNAs that have some base pairing within their 5’ untranslated regions. Mol Cell Biol. 2003;23(24):8925–8933.PubMedPubMedCentralCrossRefGoogle Scholar
  20. 20.
    Rousseau D, Gingras A, Pause A, Sonenberg N. The eIF4E-binding proteins 1 and 2 are negative regulators of cell growth. Oncogene. 1996;13(11):2415–2420.Google Scholar
  21. 21.
    Garber K. Rapamycin’s resurrection: a new way to target the cancer cell cycle. J Natl Cancer Inst. 2001;93(20):1517–1519.Google Scholar
  22. 22.
    Schuler W, Sedrani R, Cottens S, et al. SDZ RAD, a new rapamycin derivative: pharmacological properties in vitro and in vivo. Transplantation. 1997;64(1):36–42.Google Scholar
  23. 23.
    Atkins M, Hidalgo M, Stadler W, et al. Randomized phase II study of multiple dose levels of CCI-779, a novel mammalian target of rapamycin kinase inhibitor, in patients with advanced refractory renal cell carcinoma. J Clin Oncol. 2004;22(5):909–918.Google Scholar
  24. 24.
    Bhatia S, Thompson JA. Temsirolimus in patients with advanced renal cell carcinoma: an overview. Adv Ther. 2009;26(1):55–67.Google Scholar
  25. 25.
    Telfer E, McLaughlin M, Ding C, Thong K. A two-step serum-free culture system supports development of human oocytes from primordial follicles in the presence of activin. Hum Reprod. 2008; 23(5):1151–1158.Google Scholar
  26. 26.
    Xu M, Kreeger PK, Shea LD, Woodruff TK. Tissue-engineered follicles produce live, fertile offspring. Tissue Eng. 2006;12(10): 2739–2746.Google Scholar
  27. 27.
    Eppig J, Pendola F, Wigglesworth K, Pendola J. Mouse oocytes regulate metabolic cooperativity between granulosa cells and oocytes: amino acid transport. Biol Reprod. 2005;73(2):351–357.Google Scholar
  28. 28.
    Eppig J, Wigglesworth K, Pendola F. The mammalian oocyte orchestrates the rate of ovarian follicular development. Proc Natl Acad Sci U S A. 2002;99(5):2890–2894.Google Scholar
  29. 29.
    Matzuk M, Burns K, Viveiros M, Eppig J. Intercellular communication in the mammalian ovary: oocytes carry the conversation. Science. 2002;296(5576):2178–2180.Google Scholar
  30. 30.
    Cameron JL. Stress and behaviorally induced reproductive dysfunction in primates. Semin Reprod Endocrinol. 1997;15(1): 37–45.Google Scholar
  31. 31.
    Schenker JG, Meirow D, Schenker E. Stress and human reproduction. Eur J Obstet Gynecol Reprod Biol. 1992;45(1):1–8.Google Scholar
  32. 32.
    Szoltys M, Sakiewicz A, Galas J, Jaworska-Caudelier M, Kula E. Changes in follicular fluid concentrations of steroids, the pattern of steroid secretion and aromatization capability of incubated preovulatory follicular wall of rats. Reprod Biol. 2001; 1(1):42–50.Google Scholar
  33. 33.
    Neale D, Demasio K, Illuzi J, Chaiworapongsa T, Romero R, Mor G. Maternal serum of women with pre-eclampsia reduces trophoblast cell viability: evidence for an increased sensitivity to Fasmediated apoptosis. J Matern Fetal Neonatal Med. 2003;13(1): 39–44.Google Scholar
  34. 34.
    Straszewski-Chavez S, Abrahams V, Funai E, Mor G. X-linked inhibitor of apoptosis (XIAP) confers human trophoblast cell resistance to Fas-mediated apoptosis. Mol Hum Reprod. 2004; 10(1):33–41.Google Scholar
  35. 35.
    Walters K, Binnie J, Campbell B, Armstrong D, Telfer E. The effects of IGF-I on bovine follicle development and IGFBP-2 expression are dose and stage dependent. Reproduction. 2006;131(3):515–523.Google Scholar
  36. 36.
    Boulay A, Rudloff J, Ye J, et al. Dual inhibition of mTOR and estrogen receptor signaling in vitro induces cell death in models of breast cancer. Clin Cancer Res. 2005;11(14):5319–5328.Google Scholar
  37. 37.
    Chang S, Miron P, Miron A, Iglehart J. Rapamycin inhibits proliferation of estrogen-receptor-positive breast cancer cells. J Surg Res. 2007;138(1):37–44.Google Scholar
  38. 38.
    Santen R, Song R, Masamura S, et al. Adaptation to estradiol deprivation causes up-regulation of growth factor pathways and hypersensitivity to estradiol in breast cancer cells. Adv Exp Med Biol. 2008;630:19–34.Google Scholar
  39. 39.
    Yin X, Wang G, Khan-Dawood F. Requirements of phosphatidylinositol-3 kinase and mammalian target of rapamycin for estrogen-induced proliferation in uterine leiomyoma-and myometrium-derived cell lines. Am J Obstet Gynecol. 2007; 196(2):1–5.Google Scholar
  40. 40.
    Gisselsson D. Classification of chromosome segregation errors in cancer. Chromosoma. 2008;117(6):511–519.Google Scholar
  41. 41.
    Temime-Smaali N, Guittat L, Wenner T, et al. Topoisomerase IIIalpha is required for normal proliferation and telomere stability in alternative lengthening of telomeres. EMBO J. 2008;27(10): 1513–1524.PubMedPubMedCentralGoogle Scholar
  42. 42.
    Chan K, North P, Hickson I. BLM is required for faithful chromosome segregation and its localization defines a class of ultrafine anaphase bridges. EMBO J. 2007;26(14):3397–3409.Google Scholar

Copyright information

© Society for Reproductive Investigation 2012

Authors and Affiliations

  1. 1.Department of Obstetrics, Gynecology, and Reproductive Sciences, Division of Reproductive EndocrinologyYale School of MedicineNew HavenUSA
  2. 2.Program in Molecular MedicineUniversity of Massachusetts Medical SchoolWorcesterUSA

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