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Cellular and Molecular Bioengineering

, Volume 11, Issue 5, pp 435–450 | Cite as

Synergy of Paracrine Signaling During Early-Stage Mouse Ovarian Follicle Development In Vitro

  • Hong Zhou
  • Joseph T. Decker
  • Melissa M. Lemke
  • Claire E. Tomaszweski
  • Lonnie D. Shea
  • Kelly B. Arnold
  • Ariella ShikanovEmail author
Article

Abstract

Introduction

Paracrine signals, such as soluble cytokines and extracellular matrix cues, are essential for the survival and development of multicellular ovarian follicles. While it is well established that hydrogel-based culture systems successfully support the growth of late-stage follicles for fertility preservation, growing small, early-stage ovarian follicles still proves to be challenging. We hypothesized that paracrine factors secreted from neighboring follicles may be crucial for improving the survival of early-stage follicles in vitro.

Methods

To test our hypothesis, we investigated the bi-directional crosstalk of the paracrine signals, such as cell-secreted cytokines, sex hormones and transcription factors (TFs), in follicles encapsulated and cultured for 12 days in alginate in groups of five (5×) and ten (10×).

Results

The differential profiles of TF activity and secretome during folliculogenesis were analyzed using TRanscriptional Activity CEllular aRray (TRACER) and data-driven multivariate modeling approach. The mechano- and oxygen-responsive TFs, NF-κB and HIF1, exhibited a unique upregulation signature in 10× follicles. Consistently, levels of proangiogenic factors, such as VEGF-A and angiopoietin-2, were significantly higher in 10× follicles than those in 5× follicles, reaching 269.77 and 242.82 pg/mL on the last day of culture. The analysis of TRACER and secreted cytokines also revealed critical early interactions between cytokines and TFs, correlating with the observed phenotypical and functional differences between conditions.

Conclusions

We identified unique signatures of synergism during successful early-stage ovarian follicle development. These findings bring us closer to understanding of mechanisms underlying the downstream effects of interactions between the extracellular microenvironment and early-stage folliculogenesis in vitro.

Keywords

Primary ovarian follicle Synergy Paracrine signaling 

Notes

Acknowledgments

The authors would like to acknowledge the University of Virginia Center for Research in Reproduction Ligand Assay and Analysis Core (Eunice Kennedy Shriver NICHD/NIH (NCTRI) Grant P50-HD28934) for performing the hormone assays. This work was supported by The Graduate Assistance in Areas of National Need (GAANN) Fellowship to CET, R01 CA214384 to LDS and the NSF CAREER Award (#1552580) to AS.

Conflict of interest

Hong Zhou, Joseph T. Decker, Melissa M. Lemke, Claire E. Tomaszweski, Lonnie D. Shea, Kelly B. Arnold, and Ariella Shikanov declare that they have no conflicts of interest.

Ethical Standards

No human studies were carried out by the authors for this article. All animal studies were carried out in accordance with guidelines approved by The Institutional Animal Care and Use Committee (IACUC) at the University of Michigan.

Supplementary material

12195_2018_545_MOESM1_ESM.tif (10.2 mb)
Suppl. Figure 1 A. Transduction efficiency calculation. A. Representative images of broken-down follicles after transduction. B. Quantitative results of total cell numbers and luciferase-expressing cell numbers. C. Normalized luciferase production during in vitro primary follicle cultures. Linear regression was plotted as the green line with the following formula: Y = -0.033*X + 0.3628, R2 = 0.9917. The slope was not significantly different from 0 (p = 0.0582).Supplementary material 1 (TIFF 10479 kb)
12195_2018_545_MOESM2_ESM.tif (2.9 mb)
Suppl. Figure 2 Detected cytokines that showed no significant differences between 5× and 10× follicles. Data presented as mean ± SD. Supplementary material 2 (TIFF 3014 kb)
12195_2018_545_MOESM3_ESM.docx (32 kb)
Supplementary material 3 (DOCX 31.5 kb)

References

  1. 1.
    Abir, R., et al. Vascular endothelial growth factor A and its two receptors in human preantral follicles from fetuses, girls, and women. Fertil. Steril. 93(7):2337–2347, 2010.CrossRefGoogle Scholar
  2. 2.
    Agarwal, S. K., et al. Leptin antagonizes the insulin-like growth factor-I augmentation of steroidogenesis in granulosa and theca cells of the human ovary. J. Clin. Endocrinol. Metab. 84(3):1072–1076, 1999.Google Scholar
  3. 3.
    Aguado, B. A., et al. Secretome identification of immune cell factors mediating metastatic cell homing. Sci. Rep. 5:17566, 2015.CrossRefGoogle Scholar
  4. 4.
    Ahsan, S., M. Lacey, and S. A. Whitehead. Interactions between interleukin-1 beta, nitric oxide and prostaglandin E2 in the rat ovary: effects on steroidogenesis. Eur. J. Endocrinol. 137(3):293–300, 1997.CrossRefGoogle Scholar
  5. 5.
    Arnold, K. B., and A. W. Chung. Prospects from systems serology research. Immunology 153(3):279–289, 2017.CrossRefGoogle Scholar
  6. 6.
    Arnold, K. B., et al. CD4+ T cell-dependent and CD4+ T cell-independent cytokine-chemokine network changes in the immune responses of HIV-infected individuals. Sci. Signal 8(399):104, 2015.CrossRefGoogle Scholar
  7. 7.
    Baka, S., and A. Malamitsi-Puchner. Novel follicular fluid factors influencing oocyte developmental potential in IVF: a review. Reprod. Biomed. 12(4):500–506, 2006.CrossRefGoogle Scholar
  8. 8.
    Barbieri, R. L. Insulin stimulates androgen accumulation in incubations of minced porcine theca. Gynecol. Obstet. Investig. 37(4):265–269, 1994.CrossRefGoogle Scholar
  9. 9.
    Benjamini, Y., and Y. Hochberg. Controlling the false discovery rate: a practical and powerful approach to multiple testing. J. R. Stat. Soc. Ser. B 57(1):289–300, 1995.MathSciNetzbMATHGoogle Scholar
  10. 10.
    Bonnet, A., et al. Spatio-temporal gene expression profiling during in vivo early ovarian folliculogenesis: integrated transcriptomic study and molecular signature of early follicular growth. PLoS ONE 10(11):e0141482, 2015.CrossRefGoogle Scholar
  11. 11.
    Bornstein, S. R., H. Rutkowski, and I. Vrezas. Cytokines and steroidogenesis. Mol. Cell. Endocrinol. 215(1):135–141, 2004.CrossRefGoogle Scholar
  12. 12.
    Chong, I.-G., and C.-H. Jun. Performance of some variable selection methods when multicollinearity is present. Chemom. Intell. Lab. Syst. 78(1):103–112, 2005.CrossRefGoogle Scholar
  13. 13.
    Chou, C. H., and M. J. Chen. The Effect of steroid hormones on ovarian follicle development. Vitam. Horm. 107:155–175, 2018.CrossRefGoogle Scholar
  14. 14.
    Chu, S., et al. Transrepression of estrogen receptor beta signaling by nuclear factor-kappab in ovarian granulosa cells. Mol. Endocrinol. 18(8):1919–1928, 2004.CrossRefGoogle Scholar
  15. 15.
    Conti, M., et al. Role of the epidermal growth factor network in ovarian follicles. Mol. Endocrinol. 20(4):715–723, 2006.CrossRefGoogle Scholar
  16. 16.
    Decker, J. T., et al. Systems analysis of dynamic transcription factor activity identifies targets for treatment in Olaparib resistant cancer cells. Biotechnol. Bioeng. 114(9):2085–2095, 2017.CrossRefGoogle Scholar
  17. 17.
    Dull, T., et al. A third-generation lentivirus vector with a conditional packaging system. J. Virol. 72(11):8463–8471, 1998.Google Scholar
  18. 18.
    Erickson, G. F., et al. The effects of insulin and insulin-like growth factors-I and-II on estradiol production by granulosa cells of polycystic ovaries. J. Clin. Endocrinol. Metab. 70(4):894–902, 1990.CrossRefGoogle Scholar
  19. 19.
    Findlay, J. K. Peripheral and local regulators of folliculogenesis. Reprod. Fertil. Dev. 6(2):127–139, 1994.CrossRefGoogle Scholar
  20. 20.
    Findlay, J. K., et al. Recruitment and development of the follicle; the roles of the transforming growth factor-β superfamily. Mol. Cell. Endocrinol. 191(1):35–43, 2002.CrossRefGoogle Scholar
  21. 21.
    Fisher, T. E., et al. Vascular endothelial growth factor and angiopoietin production by primate follicles during culture is a function of growth rate, gonadotrophin exposure and oxygen milieu. Hum. Reprod. (Oxford, England) 28(12):3263–3270, 2013.CrossRefGoogle Scholar
  22. 22.
    Foster, R., et al. A differential cytokine expression profile is induced by highly purified human menopausal gonadotropin and recombinant follicle-stimulating hormone in a pre- and postovulatory mouse follicle culture model. Fertil. Steril. 93(5):1464–1476, 2010.CrossRefGoogle Scholar
  23. 23.
    Hahn, C., and M. A. Schwartz. Mechanotransduction in vascular physiology and atherogenesis. Nat. Rev. Mol. Cell Biol. 10(1):53–62, 2009.CrossRefGoogle Scholar
  24. 24.
    Hazzard, T. M., and R. L. Stouffer. Angiogenesis in ovarian follicular and luteal development. Baillieres Best Pract. Res. Clin. Obstet. Gynaecol. 14(6):883–900, 2000.CrossRefGoogle Scholar
  25. 25.
    Holt, J. E., et al. CXCR4/SDF1 interaction inhibits the primordial to primary follicle transition in the neonatal mouse ovary. Dev. Biol. 293(2):449–460, 2006.CrossRefGoogle Scholar
  26. 26.
    Hornick, J., et al. Multiple follicle culture supports primary follicle growth through paracrine-acting signals. Reproduction 145(1):19–32, 2013.CrossRefGoogle Scholar
  27. 27.
    Hosaka, T., et al. Disruption of forkhead transcription factor (FOXO) family members in mice reveals their functional diversification. Proc. Natl. Acad. Sci. U.S.A. 101(9):2975–2980, 2004.CrossRefGoogle Scholar
  28. 28.
    Hsueh, A. J., et al. Hormonal regulation of the differentiation of cultured ovarian granulosa cells. Endocr. Rev. 5(1):76–127, 1984.CrossRefGoogle Scholar
  29. 29.
    Ishihara, S., et al. Substrate stiffness regulates temporary NF-kappaB activation via actomyosin contractions. Exp. Cell Res. 319(19):2916–2927, 2013.CrossRefGoogle Scholar
  30. 30.
    Karin, M., and A. Lin. NF-[kappa]B at the crossroads of life and death. Nat. Immunol. 3(3):221–227, 2002.CrossRefGoogle Scholar
  31. 31.
    Kastan, M. B., et al. A mammalian cell cycle checkpoint pathway utilizing p53 and GADD45 is defective in ataxia-telangiectasia. Cell 71(4):587–597, 1992.CrossRefGoogle Scholar
  32. 32.
    Kawamura, K., et al. Paracrine regulation of the resumption of oocyte meiosis by endothelin-1. Dev. Biol. 327(1):62–70, 2009.CrossRefGoogle Scholar
  33. 33.
    Kawasaki, F., et al. The clinical role of interleukin-6 and interleukin-6 soluble receptor in human follicular fluids. Clin. Exp. Med. 3(1):27–31, 2003.CrossRefGoogle Scholar
  34. 34.
    Kim, D., J. Lee, and A. L. Johnson. Vascular endothelial growth factor and angiopoietins during hen ovarian follicle development. Gen. Comp. Endocrinol. 232:25–31, 2016.CrossRefGoogle Scholar
  35. 35.
    Knight, P. G., and C. Glister. TGF-β superfamily members and ovarian follicle development. Reproduction 132(2):191–206, 2006.CrossRefGoogle Scholar
  36. 36.
    Kreeger, P. K., et al. Regulation of mouse follicle development by follicle-stimulating hormone in a three-dimensional in vitro culture system is dependent on follicle stage and dose. Biol. Reprod. 73(5):942–950, 2005.CrossRefGoogle Scholar
  37. 37.
    Lau, K. S., et al. In vivo systems analysis identifies spatial and temporal aspects of the modulation of TNF-alpha-induced apoptosis and proliferation by MAPKs. Sci. Signal 4(165):ra16, 2011.CrossRefGoogle Scholar
  38. 38.
    Lebbe, M., et al. The steroid metabolome in the isolated ovarian follicle and its response to androgen exposure and antagonism. Endocrinology 158(5):1474–1485, 2017.CrossRefGoogle Scholar
  39. 39.
    Liu, Z., et al. Interleukin-6: an autocrine regulator of the mouse cumulus cell-oocyte complex expansion process. Endocrinology 150(7):3360–3368, 2009.CrossRefGoogle Scholar
  40. 40.
    Maisonpierre, P. C., et al. Angiopoietin-2, a natural antagonist for Tie2 that disrupts in vivo angiogenesis. Science 277(5322):55–60, 1997.CrossRefGoogle Scholar
  41. 41.
    Makanji, Y., et al. Hypoxia-mediated carbohydrate metabolism and transport promote early-stage murine follicle growth and survival. Am J Physiol Endocrinol. Metab. 306(8):E893–E903, 2014.CrossRefGoogle Scholar
  42. 42.
    Man, L., et al. Engineered endothelium provides angiogenic and paracrine stimulus to grafted human ovarian tissue. Sci. Rep. 7(1):8203, 2017.CrossRefGoogle Scholar
  43. 43.
    Mendez, U., H. Zhou, and A. Shikanov. Synthetic PEG Hydrogel for Engineering the Environment of Ovarian Follicles. In: Biomaterials for Tissue Engineering. Methods in Molecular Biology, Vol. 1758, edited by K. Chawla. New York: Humana Press, 2018, pp. 115–128.CrossRefGoogle Scholar
  44. 44.
    Molineux, G., et al. The effects on hematopoiesis of recombinant stem cell factor (ligand for c-kit) administered in vivo to mice either alone or in combination with granulocyte colony-stimulating factor. Blood 78(4):961–966, 1991.Google Scholar
  45. 45.
    Motro, B., et al. Pattern of interleukin 6 gene expression in vivo suggests a role for this cytokine in angiogenesis. Proc Natl Acad Sci USA 87(8):3092–3096, 1990.CrossRefGoogle Scholar
  46. 46.
    Murray, S., et al. Regulation of granulosa cell-derived ovarian metalloproteinase inhibitor(s) by prolactin. J. Reprod. Fertil. 107(1):103–108, 1996.CrossRefGoogle Scholar
  47. 47.
    Nilsson, E., and M. K. Skinner. Cellular interactions that control primordial follicle development and folliculogenesis. J. Soc. Gynecol. Investig. 8:S17–S20, 2001.CrossRefGoogle Scholar
  48. 48.
    O’Dwyer, D. N., et al. The peripheral blood proteome signature of idiopathic pulmonary fibrosis is distinct from normal and is associated with novel immunological processes. Sci. Rep. 7:46560, 2017.CrossRefGoogle Scholar
  49. 49.
    Park, J.-Y., et al. EGF-like growth factors as mediators of LH action in the ovulatory follicle. Science 303(5658):682–684, 2004.CrossRefGoogle Scholar
  50. 50.
    Penalver Bernabe, B., et al. Dynamic transcription factor activity networks in response to independently altered mechanical and adhesive microenvironmental cues. Integr. Biol. (Camb) 8(8):844–860, 2016.CrossRefGoogle Scholar
  51. 51.
    Sabbaghi, M., et al. IL-17A concentration of seminal plasma and follicular fluid in infertile men and women with various clinical diagnoses. Immunol. Invest. 43(7):617–626, 2014.MathSciNetCrossRefGoogle Scholar
  52. 52.
    Schneider, C. A., W. S. Rasband, and K. W. Eliceiri. NIH Image to ImageJ: 25 years of image analysis. Nat. Methods 9(7):671–675, 2012.CrossRefGoogle Scholar
  53. 53.
    Shikanov, A., et al. Interpenetrating fibrin-alginate matrices for in vitro ovarian follicle development. Biomaterials 30(29):5476–5485, 2009.CrossRefGoogle Scholar
  54. 54.
    Shikanov, A., et al. A method for ovarian follicle encapsulation and culture in a proteolytically degradable 3 dimensional system. J. Vis. Exp. 49:2695, 2011.Google Scholar
  55. 55.
    Shikanov, A., et al. Hydrogel network design using multifunctional macromers to coordinate tissue maturation in ovarian follicle culture. Biomaterials 32(10):2524–2531, 2011.CrossRefGoogle Scholar
  56. 56.
    Shweiki, D., et al. Patterns of expression of vascular endothelial growth factor (VEGF) and VEGF receptors in mice suggest a role in hormonally regulated angiogenesis. J. Clin. Investig. 91(5):2235–2243, 1993.CrossRefGoogle Scholar
  57. 57.
    Skaznik-Wikiel, M. E., et al. Granulocyte colony-stimulating factor in conjunction with vascular endothelial growth factor maintains primordial follicle numbers in transplanted mouse ovaries. Fertil. Steril. 95(4):1405–1409, 2011.CrossRefGoogle Scholar
  58. 58.
    Smyth, G. K., et al. LIMMA: linear models for microarray data. In: Bioinformatics and Computational Biology Solutions Using R and Bioconductor, edited by R. Gentleman, V. Carey, S. Dudoit, R. Irizarry, and W. Huber. New York: Springer, 2005, pp. 397–420.CrossRefGoogle Scholar
  59. 59.
    Srisuma, S., et al. Identification of genes promoting angiogenesis in mouse lung by transcriptional profiling. Am. J. Respir. Cell Mol. Biol. 29(2):172–179, 2003.CrossRefGoogle Scholar
  60. 60.
    Stouffer, R. L., et al. Regulation and action of angiogenic factors in the primate ovary. Arch. Med. Res. 32(6):567–575, 2001.CrossRefGoogle Scholar
  61. 61.
    Tagler, D., et al. Embryonic fibroblasts enable the culture of primary ovarian follicles within alginate hydrogels. Tissue Eng. Part A 18(11–12):1229–1238, 2012.CrossRefGoogle Scholar
  62. 62.
    Tamadon, A., et al. Efficient biomaterials for tissue engineering of female reproductive organs. Tissue Eng. Regener. Med. 13(5):447–454, 2016.CrossRefGoogle Scholar
  63. 63.
    Tamanini, C., and M. De Ambrogi. Angiogenesis in developing follicle and corpus luteum. Reprod. Domest. Anim. 39(4):206–216, 2004.CrossRefGoogle Scholar
  64. 64.
    Taniguchi, T. Cytokine signaling through nonreceptor protein tyrosine kinases. Science 268(5208):251–255, 1995.CrossRefGoogle Scholar
  65. 65.
    Tingen, C. M., et al. A macrophage and theca cell-enriched stromal cell population influences growth and survival of immature murine follicles in vitro. Reproduction 141(6):809–820, 2011.CrossRefGoogle Scholar
  66. 66.
    Trau, H. A., et al. Prostaglandin E2 and vascular endothelial growth factor A mediate angiogenesis of human ovarian follicular endothelial cells. Hum. Reprod. 31(2):436–444, 2016.Google Scholar
  67. 67.
    van den Hurk, R., and J. Zhao. Formation of mammalian oocytes and their growth, differentiation and maturation within ovarian follicles. Theriogenology 63(6):1717–1751, 2005.CrossRefGoogle Scholar
  68. 68.
    Viedt, C., et al. Monocyte chemoattractant protein-1 induces proliferation and interleukin-6 production in human smooth muscle cells by differential activation of nuclear factor-kappaB and activator protein-1. Arterioscler. Thromb. Vasc. Biol. 22(6):914–920, 2002.CrossRefGoogle Scholar
  69. 69.
    Wang, Y., S. Chan, and B. K. Tsang. Involvement of inhibitory nuclear factor-kappaB (NFkappaB)-independent NFkappaB activation in the gonadotropic regulation of X-linked inhibitor of apoptosis expression during ovarian follicular development in vitro. Endocrinology 143(7):2732–2740, 2002.CrossRefGoogle Scholar
  70. 70.
    Weiss, M. S., et al. Dynamic, large-scale profiling of transcription factor activity from live cells in 3D culture. PLoS ONE 5(11):e14026, 2010.CrossRefGoogle Scholar
  71. 71.
    Weiss, M. S., et al. Dynamic transcription factor activity and networks during ErbB2 breast oncogenesis and targeted therapy. Integr. Biol. (Camb) 6(12):1170–1182, 2014.CrossRefGoogle Scholar
  72. 72.
    West, E. R., et al. Physical properties of alginate hydrogels and their effects on in vitro follicle development. Biomaterials 28(30):4439–4448, 2007.CrossRefGoogle Scholar
  73. 73.
    Wigglesworth, K., et al. Transcriptomic diversification of developing cumulus and mural granulosa cells in mouse ovarian follicles. Biol. Reprod. 92(1):23, 2015.CrossRefGoogle Scholar
  74. 74.
    Xu, M., et al. Tissue-engineered follicles produce live, fertile offspring. Tissue Eng. 12(10):2739–2746, 2006.CrossRefGoogle Scholar
  75. 75.
    Xu, M., et al. Encapsulated three-dimensional culture supports development of nonhuman primate secondary follicles. Biol. Reprod. 81(3):587–594, 2009.CrossRefGoogle Scholar
  76. 76.
    Xu, M., et al. Secondary follicle growth and oocyte maturation by culture in alginate hydrogel following cryopreservation of the ovary or individual follicles. Biotechnol. Bioeng. 103(2):378–386, 2009.CrossRefGoogle Scholar
  77. 77.
    Xu, J., et al. Secondary follicle growth and oocyte maturation during encapsulated three-dimensional culture in rhesus monkeys: effects of gonadotrophins, oxygen and fetuin. Hum. Reprod. 26(5):1061–1072, 2011.CrossRefGoogle Scholar
  78. 78.
    Yeung, F., et al. Modulation of NF-κB-dependent transcription and cell survival by the SIRT1 deacetylase. EMBO J 23(12):2369–2380, 2004.CrossRefGoogle Scholar
  79. 79.
    Young, J. M., and A. S. McNeilly. Theca: the forgotten cell of the ovarian follicle. Reproduction 140(4):489–504, 2010.CrossRefGoogle Scholar
  80. 80.
    Zimmermann, R. C., et al. Vascular endothelial growth factor receptor 2–mediated angiogenesis is essential for gonadotropin-dependent follicle development. J. Clin. Investig. 112(5):659, 2003.CrossRefGoogle Scholar

Copyright information

© Biomedical Engineering Society 2018

Authors and Affiliations

  • Hong Zhou
    • 1
  • Joseph T. Decker
    • 1
  • Melissa M. Lemke
    • 1
  • Claire E. Tomaszweski
    • 1
  • Lonnie D. Shea
    • 1
    • 2
  • Kelly B. Arnold
    • 1
  • Ariella Shikanov
    • 1
    • 3
    • 4
    Email author
  1. 1.Department of Biomedical EngineeringUniversity of MichiganAnn ArborUSA
  2. 2.Department of Chemical EngineeringUniversity of MichiganAnn ArborUSA
  3. 3.Department of Macromolecular Science and EngineeringUniversity of MichiganAnn ArborUSA
  4. 4.Ann ArborUSA

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