Cellular Mechanisms of Chemotherapy-induced Male Infertility: Stem Cell or Niche?

化疗引起的男性不育的细胞机制：干细胞还是利基？

• 批准号: 9085497
• 负责人: Kyle Edwin Orwig
• 金额: $ 6.14万
• 依托单位: MAGEE-WOMEN'S RES INST AND FOUNDATION
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-06-01 至 2019-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9085497
• 关键词: Address; Age; Anatomy; Animal Model; Autologous; Biological Preservation; Biopsy; Bone Marrow Transplantation; Busulfan; Cancer Survivor; Cancer Survivorship; Cell Culture Techniques; Cell Survival; Cell Therapy; Cell physiology; Cell surface; Cells; Chemotherapy-Oncologic Procedure; Clinic; Clinical; Cyclophosphamide; Data; Defect; Diagnosis; Dose; Endocrine; Environment; Exposure to; Fertility; Follicle Stimulating Hormone; Foundations; Freezing; Future; Gene Expression; Gonadal structure; Health; Hematopoietic Stem Cell Transplantation; High Dose Chemotherapy; Human; Hypothalamic structure; Infertility; Knowledge; Lead; Life; Luteinizing Hormone; Macaca mulatta; Male Infertility; Malignant Neoplasms; Mediating; Micromanipulation; Mind; Modeling; Monkeys; Morphology; Natural regeneration; Non-Malignant; Nuclear; Nude Mice; Patients; Physiology; Pituitary Gland; Plants; Population; Precocious Puberty; Pregnancy; Primates; Production; Proliferating; Property; Puberty; RNA Sequences; Radiation; Radiation therapy; Recovery; Regulation; Relative (related person); Research; Research Institute; Rodent; Safety; Schedule; Signal Pathway; Signal Transduction; Spermatogenesis; Spermatogonia; Staining method; Stains; Stem cell transplant; Stem cells; Techniques; Technology; Testicular Tissue; Testing; Testis; Testosterone; Therapeutic; Therapeutic Intervention; Tissues; Toxic effect; Translating; Transplantation; United States; Walkers; Whole-Body Irradiation; Woman; Xenograft procedure; base; boys; cancer therapy; chemotherapy; childhood cancer survivor; clinically relevant; conditioning; cytotoxic; experience; high risk; innovation; insight; leydig interstitial cell; male; men; nonhuman primate; novel; patient population; pre-clinical; preclinical study; prepuberty; programs; regenerative; sertoli cell; sperm cell; stem cell niche; stem cell population; stem cell technology; stem cell therapy; transcriptome sequencing

DESCRIPTION (provided by applicant): Spermatogonial stem cells (SSCs) are at the foundation of spermatogenesis and may have application for treating some cases of male infertility. High dose chemotherapy treatments for cancer and other non-malignant conditions can cause azoospermia, which may be due to a depletion of the SSC pool. If that is the case and the testicular environment (SSC niche) is functionally intact, then SSC transplantation may constitute a cell based therapy for restoring fertility in male cancer survivors. Theoretically, SSCs can be isolated via biopsy and cryopreserved prior to cancer treatment and then reintroduced into the testis after cure to regenerate spermatogenesis. The proof in principle for this approach is already established in several animal models, including recent progress from our lab in primates. Therefore, it is tempting to speculate that SSC transplantation might be translated to the human fertility clinic. This option may be particularly appropriate for prepubert boys who are not yet producing sperm and have no options to preserve their future fertility. With this in mind, several academic centers around the world, including our Fertility Preservation Program in Pittsburgh, are cryopreserving testicular tissues for boys in anticipation that SSCs can be used in the future to restore fertility. Pre-clinical studies are critically needed to demonstrate the feasibility of SSC transplantation in a model that is relevant to human anatomy and physiology as well as the target prepubertal patient population. In this application, we will treat prepubertal rhesus macaques with a clinically relevant high-dose alkylating chemotherapy regimen (BuCy2) that is expected to cause azoospermia. The SSC pool in nonhuman primate and human testes is comprised of Adark and Apale spermatogonia, but the relative regenerative potential of these morphologically distinct spermatogonial subtypes are not known. In Aim 1 we will evaluate and compare the stem cell properties of Adark and Apale spermatogonia, which will impact strategies to manipulate these cells for therapeutic purposes. Aims 2 will examine the effects of BuCy2 on the stem cell pool and somatic/endocrine environment (stem cell niche) of prepubertal testes. Aim 3 will model the clinical scenario of the prepubertal patient that is at high risk for azoospermia due to BuCy2 conditioning prior to hematopoietic stem cell transplantation. In that model, we will perform autologous SSC transplantation from frozen/thawed testicular cells. Fundamental knowledge generated in aims 1 and 2 could lead to novel SSC enrichment and niche therapy strategies that will be applied in Aim 3 to enhance spermatogenesis from endogenous or transplanted SSCs.

描述（由申请人提供）：精原干细胞（SSC）是精子发生的基础，可用于治疗某些男性不育症。针对癌症和其他非恶性疾病的高剂量化疗可能会导致无精子症，这可能是由于 SSC 库耗尽所致。如果情况确实如此，并且睾丸环境（SSC 生态位）功能完整，那么 SSC 移植可能构成一种基于细胞的疗法，用于恢复男性癌症幸存者的生育能力。理论上，SSCs可以在癌症治疗前通过活检分离并冷冻保存，然后在治愈后重新引入睾丸以再生精子发生。这种方法的原则证明已经在几种动物模型中建立，包括我们实验室在灵长类动物身上取得的最新进展。因此，人们很容易推测 SSC 移植可能会转化为人类生育诊所。此选项可能特别适合尚未产生精子且没有选择保留未来生育能力的青春期前男孩。考虑到这一点，世界各地的多个学术中心，包括我们位于匹兹堡的生育力保存项目，正在冷冻保存男孩的睾丸组织，以期将来可以使用 SSC 来恢复生育力。迫切需要临床前研究来证明在与人体解剖学和生理学以及目标青春期前患者群体相关的模型中进行 SSC 移植的可行性。在此应用中，我们将使用临床相关的高剂量烷化化疗方案 (BuCy2) 治疗青春期前的恒河猴，该方案预计会导致无精子症。非人灵长类动物和人类睾丸中的 SSC 池由 Adark 和 Apale 精原细胞组成，但这些形态上不同的精原细胞亚型的相对再生潜力尚不清楚。在目标 1 中，我们将评估和比较 Adark 和 Apale 精原细胞的干细胞特性，这将影响操纵这些细胞用于治疗目的的策略。目标 2 将检查 BuCy2 对青春期前睾丸干细胞池和体细胞/内分泌环境（干细胞生态位）的影响。目标 3 将模拟青春期前患者的临床情况，即 造血干细胞移植前进行 BuCy2 调节导致无精子症的高风险。在该模型中，我们将从冷冻/解冻的睾丸细胞中进行自体 SSC 移植。目标 1 和 2 中产生的基础知识可能会导致新的 SSC 富集和利基治疗策略，这些策略将应用于目标 3，以增强内源性或移植 SSC 的精子发生。