Oocyte-derived R-spondin2 as a Follicle Stimulating Hormone


Principal Investigator: AARON JW HSUEH
Abstract: DESCRIPTION (provided by applicant): Oocyte-derived R-spondin2 as a Follicle Stimulating Hormone Abstract: The Wingless (Wnt) signaling pathway is essential for cell proliferation from flies to mammals. Although multiple Wnt ligands and their cognate Frizzled receptors are expressed in ovarian somatic cells, there is no report demonstrating the ability of Wnt ligands in the promotion of ovarian follicle growth. Based on bioinformatic and in situ hybridization analyses, we demonstrated the exclusive expression of R-spondin2 in oocytes of primary and more advanced follicles but not in oocytes of dormant primordial follicles. There are four R-spondin paralogous genes (R-spondin1-4) in the human genome and these secreted proteins have been found to be potent growth factors for adult stem cells in gastrointestinal organs and hair follicles by serving as co-ligands for the Wnt signaling pathway essential for cell proliferation. Recent studies further demonstrated the role of LGR4/5/6 proteins as the cognate receptors for R-spondin ligands. Our preliminary data demonstrated the ability of R-spondin2 to synergize with Wnt ligands in the activation of the Wnt signaling pathway in cultured ovarian somatic cells. R-spondin2 treatment also promoted ovarian follicle development in cultured ovarian explants. Unlike FSH, R-spondin2 did not activate the cAMP-protein kinase A pathway and showed additive stimulation of ovarian cell proliferation together with FSH. We further generated a secreted R- spondin agonist, R-spondin1-Fc, by fusing R-spondin1 cDNA with that for the Fc domain of IgG. We showed the ability of R-spondin1-Fc to promote the growth of primary follicles to the secondary stage in neonatal mice in vivo. Pre-treatment with R-spondin1-Fc led to the induction of early antral follicles capable of responding to sequential eCG and hCG treatment, leading to the generation of mature oocytes. These oocytes could be fertilized in vitro and developed into blastocysts. Although FSH has been used extensively for the treatment of female infertility, a sub-population of patients showed low FSH responses and has no alternative therapeutic options. The present R21 proposal will first demonstrate the ovarian expression of R-spondin2 ligand and LGR4/5/6 receptor proteins, together with the mediatory role of ovarian LGR4/5/6 receptors. This will be followed by the generation of recombinant R-spondin2 to stimulate early follicle growth in neonatal wild type mice. Following the promotion of early ovarian follicle development using R-spondin2, mice will be treated sequentially with eCG and hCG to stimulate the final phase of follicle development and to derive mature oocytes for fertilization and the derivation of pups. We will then use a rodent model of low FSH responsiveness (FSH receptor haploinsufficient heterozygous mutants) to test the ability of R-spondin2 in the promotion of early follicle development for subsequent gonadotropin stimulation and the generation of mature oocytes. We will evaluate epigenetic changes, mitochondrial integrity and early embryonic development of mature oocytes to insure the efficacy and safety of the present treatment regimen. This will be followed by the derivation of healthy pups as the basis for future clinical application using R-spondin2 to treat infertile patients with low FSH responses.
Funding Period: 2012-08-10 - 2014-12-30
more information: NIH RePORT

Top Publications

  1. pmc Oocyte-derived R-spondin2 promotes ovarian follicle development
    Yuan Cheng
    Program of Reproductive and Stem Cell Biology, Stanford University School of Medicine, Stanford, CA 94305 5317, USA
    FASEB J 27:2175-84. 2013

Detail Information


  1. pmc Oocyte-derived R-spondin2 promotes ovarian follicle development
    Yuan Cheng
    Program of Reproductive and Stem Cell Biology, Stanford University School of Medicine, Stanford, CA 94305 5317, USA
    FASEB J 27:2175-84. 2013

Research Grants30

  1. Molecular and Clinical Pharmacology of Retinopathy of Prematurity
    Jacob V Aranda; Fiscal Year: 2013
    ..The NYPD-PRC will surely elevate the level of scientific inquiry on molecular and clinical pharmacology of ROP to hasten its prevention and avert life-long blindness. ..
    John H Nilson; Fiscal Year: 2013
    ..Understanding how FSH signals to direct follicular maturation can translate into safer and more effective treatments of infertility and early pregnancy loss as well as new approaches for contraceptive drugs. ..
  3. The Aging Pituitary-Gonadal Axis
    George R Bousfield; Fiscal Year: 2013
    ..While currently available preparations work well in young women, they become increasingly ineffective in older women, requiring higher doses and prolonged administration yet producing fewer oocytes. ..
  4. Stanford University Center for Reproductive and Stem Cell biology
    Margaret T Fuller; Fiscal Year: 2013
    ..abstract_text> ..
  5. Harvard Reproductive Endocrine Sciences Center
    William Francis Crowley; Fiscal Year: 2013
    ..abstract_text> ..
  6. Rocky Mountain Regional Center of Excellence or Biodefense and Emerging Infectiou
    John T Belisle; Fiscal Year: 2013
    ..abstract_text> ..
  7. Center for the Study of Reproductive Biology and Women's Health
    Jeffrey W Pollard; Fiscal Year: 2013
    ..He holds several senior administrative appointments in the College of Medicine and is well able to administer the proposed SCCPIR internally and to enable effective interactions with other SCCPIRs. ..
  8. Developmental Gene-Environment Interactions and Premature Ovarian Failure
    Ulrike Luderer; Fiscal Year: 2013
    ..Understanding these mechanisms is important to developing interventions for the prevention of premature ovarian failure. ..
  9. Lineage tracing of germline stem cell differentiation in adult ovaries
    Ning Wang; Fiscal Year: 2013
  10. Baylor Intellectual and Developmental Disabilities Research Center
    Huda Y Zoghbi; Fiscal Year: 2013
    ..abstract_text> ..
  11. Cellular/Molecular Pathophysiology of Intellectual and Developmental Disabilities
    Stuart A Lipton; Fiscal Year: 2013
    ..The CORE supports administration, statistics, tissue culture, and crystallography/modeling of NMDAR subunits and functional sites, all critical to the proposed Projects. ..
    Martin M Matzuk; Fiscal Year: 2013
  13. Center for Reproductive Science and Medicine
    Pamela L Mellon; Fiscal Year: 2013
    ..The SCCPIR Human Ovary Tissue Bank provides tissue to NIH-funded investigators nation-wide. ..
  14. Mechanisms of phthalate-induced ovarian follicle toxicity
    ZELIEANN RIVERA CRAIG; Fiscal Year: 2013
    ..Finally, results from this project will also contribute to research aimed at the identification of new treatments to reverse female infertility caused by environmental chemical exposure. ..
  15. ApoE Receptor Biology and Neurodegeneration
    Mary Jo Ladu; Fiscal Year: 2013
    ..This Program will thus provide new and valuable information about how apoE and apoE receptors affect the pathogenesis of Alzheimer's disease. ..
  16. Origins and Biological Consequences of Human Infertility
    Linda C Giudice; Fiscal Year: 2013
    ..abstract_text> ..