In boys and young men, cancer therapeutic agents result in prolonged or permanent azoospermia. Preservation of these patients'fertility requires treatments that maintain endogenous stem cells and their differentiation and, in cases of total stem cell loss, autologous spermatogonial transplantation. In a rat model we proved that in-radiation or chemotherapy produced azoospermia by impairing the testicular somatic function resulting in a failure in germ cell differentiation. However, transient suppression of testosterone (T) and FSH with GnRH-antagonist (-ant) after radiation maintained or restored spermatogenesis and fertility in these rats. Similarly, in irradiated mice and monkeys, GnRH-ant treatment improved recovery of spermatogenesis from both endogenous and transplanted spermatogonial stem cells (SSC). We hypothesize that in irradiated monkeys and mice, T-suppression is most important for enhancing colonization by transplanted SSC but that maintenance of FSH levels would promote their development. Using an irradiated rhesus macaque model relevant to pediatric cancer patients, we will assess the effects of FSH maintenance during hormone suppression on spermatogenic recovery from transplanted cryopreserved germ cells from prepubertal monkeys, as well as from endogenous SSC. To more clearly define the role of T and FSH and extend results to chemotherapy, we will assess the colonization and/or spermatogenic recovery from SSC in prepubertal monkey and mouse testis cells transplanted into irradiated and busulfan- treated mouse recipient testes during the GnRH treatment, independently modulating T and FSH. Since there appears to be progressive loss of putative stem spermatogonia shortly after radiation in these two species, we hypothesize that these are indeed functional stem cells and that hormone suppression does indeed stimulate the recovery of spermatogenesis, partly by preventing the progressive loss of these surviving SSC. We will characterize these surviving putative SSC in irradiated monkeys and mice using kinetics and SSC markers, determine their viability and functionality by transplantation into recipient mouse testes, and test whether hormone suppression of the donor prevents the loss of those cells.

Public Health Relevance

The fulfillment of the aims in this project will determine how hormone suppression can best be used to optimize the strategy for the restoration of male fertility in prepubertal cancer patients by subsequent autologous germ cell transplantation. The increased efficiency of the procedure will help make presentation of the testis tissue before cancer therapy a common and reliable practice for later transplantation.

National Institute of Health (NIH)
Eunice Kennedy Shriver National Institute of Child Health & Human Development (NICHD)
Research Program Projects (P01)
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Special Emphasis Panel (ZHD1)
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Magee-Women's Research Institute and Foundation
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Clark, Amander T; Orwig, Kyle E (2018) Editorial. Stem Cell Res 29:179
Shetty, G; Wu, Z; Lam, T N A et al. (2018) Effect of hormone modulations on donor-derived spermatogenesis or colonization after syngeneic and xenotransplantation in mice. Andrology :
Shetty, Gunapala; Mitchell, Jennifer M; Lam, Truong Nguyen Anh et al. (2018) Donor spermatogenesis in de novo formed seminiferous tubules from transplanted testicular cells in rhesus monkey testis. Hum Reprod 33:2249-2255
Sosa, Enrique; Chen, Di; Rojas, Ernesto J et al. (2018) Differentiation of primate primordial germ cell-like cells following transplantation into the adult gonadal niche. Nat Commun 9:5339
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Gassei, Kathrin; Sheng, Yi; Fayomi, Adetunji et al. (2017) DDX4-EGFP transgenic rat model for the study of germline development and spermatogenesis. Biol Reprod 96:707-719
Sosa, Enrique; Kim, Rachel; Rojas, Ernesto J et al. (2017) An integration-free, virus-free rhesus macaque induced pluripotent stem cell line (riPSC90) from embryonic fibroblasts. Stem Cell Res 21:5-8
Clark, Amander T; Gkountela, Sofia; Chen, Di et al. (2017) Primate Primordial Germ Cells Acquire Transplantation Potential by Carnegie Stage 23. Stem Cell Reports 9:329-341
Chen, Di; Gell, Joanna J; Tao, Yu et al. (2017) Modeling human infertility with pluripotent stem cells. Stem Cell Res 21:187-192

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