Derivation of Functional Spermatogonia Stem Cells from Rhesus Macaque iPSCs

摘要 目前，全世界约15%的夫妇和美国约12%的男性不育或不育。一些人 这种不育症的根本原因在于基因缺陷，但也有其他原因是由于暴露在工业和环境中 毒物、伤害或几乎总是导致不孕不育的烷化化疗等医学治疗， 尤其是在男人身上。男性青春期前癌症患者尤其危险，因为他们无法提供 癌症治疗前的精液样本。青春期前儿童癌症患者的生育力保留 一个新兴且不断发展的领域。然而，治疗选择有限。自1980年以来，辅助生殖 技术(ART)已在世界范围内被接受，胞浆内单精子注射(ICSI)自那以来 帮助有严重不孕症的夫妇怀孕。然而，这些先进的技术需要 即使是少量功能性配子的产生。对于那些因非遗传因素而不育的个体 根本原因是无法产生配子，如儿科癌症幸存者，他们的不孕不育没有治愈方法。 随着儿童癌症，包括睾丸癌和生殖细胞癌的发病率持续上升，有一种 需要开发基于干细胞的新疗法来治疗男性因素不育症患者 通过药物治疗不育的。患者特异性多能干细胞是未来的一个潜在来源 通过在体外产生生殖系前体细胞，如精原干细胞(SSCs)，恢复生育能力 移植后体内恢复生育能力。这种治疗方法将允许不育男性产生 无需使用卵胞浆内单精子注射等先进生殖技术，即可与配偶生育后代。我们的团队 首创将自体和供体恒河猴SSCs移植到受体恒河猴睾丸中的能力 移居睾丸并恢复生育能力。此外，我们的团队是第一个从 人诱导多能干细胞(IPSCs)。在这里，我们假设NHP IPSC派生的SSC可以是 移植到无菌的自体受体睾丸中以恢复生育能力。此外，我们假设精子 从移植的接受者中产生的胚胎可以产生健康的胚胎和后代。这项提案低于票面价值- 干细胞再生医学动物模型的改进(R01)将确定，使用 非人灵长类干细胞模型，ipscs能否用于治疗男性因素不育。

Summary Currently, ~15% of couples worldwide and ~12% of men in the United States are subfertile or infertile. Some root causes for this infertility lie in genetic defects, but others are due to exposure to industrial and environmental toxicants, injury, or medical treatments such as alkylating chemotherapies which almost always result in sterility, especially in men. Male pre-pubertal cancer patients are especially at risk since they are unable to provide a semen sample prior to their cancer treatments. Fertility preservation in pre-pubertal pediatric cancer patients is an emerging and ever-evolving field. However treatment options are limited. Since 1980, Assisted Reproductive Technology (ART) has gained worldwide acceptance and Intracytoplasmic Sperm Injection (ICSI) has since aided couples with severe infertility to achieve pregnancies. However, these advanced techniques require the production of even a small amount of functional gametes. For those individuals rendered sterile by non-genetic root causes unable to produce gametes, such as pediatric cancer survivors, there are no cures for their infertility. As the incidences of pediatric cancers, including testicular and germ cell cancers, continue to rise, there is a need for the development of novel stem-cell based therapies to treat male factor infertility in patients rendered sterile by medical treatments. Patient-specific pluripotent stem cells represent one future potential source to restore fertility by generating germline precursors such as spermatogonial stem cells (SSCs) in vitro that can restore fertility in vivo following transplantation. This therapeutic approach would allow infertile men to produce offspring with their partners without having to use advanced reproductive technologies such as ICSI. Our team has pioneered the ability to transplant autologous and donor rhesus SSCs into recipient rhesus testes to re- colonize the testis and restore fertility. Additionally, our group was the first group to derive SSC-like cells from human induced pluripotent stem cells (iPSCs). Here, we hypothesize that NHP iPSC-derived SSCs can be transplanted into sterilized autologous recipient testes to restore fertility. Furthermore, we hypothesize that sperm generated from transplanted recipients can produce healthy embryos and offspring. This proposal under PAR- 16-093 Improvement of Animal Models for Stem Cell-based Regenerative Medicine (R01) will determine, using a non-human primate stem cell model, whether iPSCs can be utilized to treat male factor infertility.