Apoptotic dysregulation in male infertility

男性不育中的细胞凋亡失调

• 批准号: 10391711
• 负责人: Kaitlyn Ann Webster
• 金额: $ 3.43万
• 依托单位: HARVARD MEDICAL SCHOOL
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-01 至 2022-01-02
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10391711
• 关键词: Address; Adolescent; Adult; Advisory Committees; Affect; Age; Apoptosis; Apoptotic; Automobile Driving; BCL2 gene; Bax protein; Biological Assay; Biometry; Cell Cycle Stage; Cell Death; Cell Differentiation process; Cell Survival; Cell Transplantation; Cells; Cessation of life; Child; Continuing Education; Coupled; Couples; Cues; Data Analyses; Dependence; Development; Developmental Biology; Discipline; Disease; Disease model; Embryo; Embryonic Development; Endocrinology; Environmental Exposure; Epidemiology; Equilibrium; Etiology; Event; Excision; Experimental Designs; Exposure to; Faculty; Failure; Fellowship; Female; Fertility; Flow Cytometry; Genetic; Genetic Diseases; Genetic Models; Genitourinary system; Germ Cells; Goals; Gonadal Steroid Hormones; Gonadal structure; Histologic; Hormonal; Human; Image; Immunohistochemistry; Impairment; Incidence; Infertility; Inhibition of Apoptosis; Knock-out; Laboratories; Lead; Long-Term Effects; Maintenance; Male Infertility; Male Sterility; Mammals; Maps; Measures; Mediating; Mexican; Mitochondria; Modeling; Molecular Biology; Morphogenesis; Mus; Ovary; Pathway interactions; Phase; Physiological; Pituitary Gland; Process; Production; Protein Analysis; Protein Family; Proteins; Public Health; Public Health Schools; Quantitative Trait Loci; Recovery; Regulation; Reproductive Health; Reproductive Sciences; Research; Research Support; Research Training; Resistance; Sex Differentiation; Sexual Development; Signal Transduction; Spermatocytes; Spermatogenesis; Sterility; Structure; Structure of paramesonephric duct; Testing; Testis; Time; Tissues; Toxic Environmental Substances; Toxicant exposure; Training; VDAC1 gene; Zebrafish; beta catenin; bridge program; cancer therapy; cost; cytochrome c; cytotoxic; egg; environment related cancer; fertility preservation; fetal; hormonal signals; hypothalamic pituitary gonadal axis; insight; irradiation; male; male fertility; men; mullerian-inhibiting hormone; novel; predictive test; prepuberty; prevent; progenitor; regenerative; reproductive; reproductive outcome; reproductive tract; response; response to injury; responsible research conduct; sex; sex development disorder; spatiotemporal; sperm cell; stressor; toxicant

Project Summary/Abstract Across species, sexual development and fertility are dependent on cell death events. In developing ovaries and testes in mammals, massive death of germ cells (precursors of sperm and eggs) occurs during a fetal attrition event that is presumed to prevent overproliferation and select for highest quality cells. Cell death is also observed in males during embryonic regression of structures that form the female reproductive tract, and morphogenesis of the pituitary gland, which regulates gonadal sex hormone production. Surprisingly, inhibition of apoptosis results in male-specific infertility in mice, but it is not understood why this occurs or how cell death is regulated throughout male sexual development. The goal of my research is to understand regulation of apoptosis along the hypothalamic-pituitary-gonadal (HPG) axis throughout male sexual development, and why disruption of apoptosis causes infertility. Specifically, this proposal aims to map apoptotic sensitivity in normal testicular germ cells, and to find evidence of apoptotic misregulation in reproductive tissues in infertile males. In Aim 1, the apoptotic vulnerability of germ cells at different stages of development and differentiation will be determined using functional cell death-predictive assays and testicular irradiation to induce germ cell death. In Aim 2, the requirement of apoptosis in male sex differentiation will be investigated by analyzing embryonic reproductive tract development and adult HPG axis signaling in genetic models of apoptosis insufficiency. Testicular germ cell transplantation will be used to establish contribution of germline vs. somatic testicular cell apoptosis dysregulation to male infertility. These findings will support identification of targeted treatments for disorders of sexual development or cytotoxic exposures affecting developing children, which lead to poor reproductive outcomes as adults. The training plan for the proposed fellowship will further develop expertise in molecular and developmental biology approaches, and address conceptual and technical inexperience in other disciplines of reproductive science, including endocrinology and epidemiology. It will also provide the opportunity to move from zebrafish to mice as a model of disease and development. This will be achieved by continuing education in biostatistics, training with a co-sponsor, and utilizing an interdisciplinary scientific advisory committee to oversee experimental design and data analysis. Responsible conduct of research training will also be undertaken in Year 1. The Harvard School of Public Health, which is committed to the multi-faceted study of reproductive health and infertility, will strongly support this research and training plan with imaging and analytical facilities as well as oversight of established faculty in the field. The laboratory of Kristopher Sarosiek, which excels at functional assays of apoptosis, will enable development of a unique and rigorous research program that bridges sexual development and the molecular biology of cell death in order to uncover mechanisms driving male infertility.

项目概要/摘要 在物种中，性发育和生育能力取决于细胞死亡事件。开发中 在哺乳动物的卵巢和睾丸中，生殖细胞（精子和卵子的前体）大量死亡 胎儿消耗事件被认为可以防止过度增殖并选择最高质量的细胞。细胞死亡是 在形成女性生殖道的结构的胚胎退化期间也在男性中观察到，并且 垂体的形态发生，调节性激素的产生。令人惊讶的是，抑制 细胞凋亡导致小鼠雄性特异性不育，但尚不清楚为什么会发生这种情况或细胞死亡是如何发生的 在整个男性性发育过程中受到调节。我的研究目标是了解监管 整个男性性发育过程中下丘脑-垂体-性腺 (HPG) 轴的细胞凋亡及其原因 细胞凋亡的破坏会导致不孕。具体来说，该提案旨在绘制正常细胞中的细胞凋亡敏感性 睾丸生殖细胞，并寻找不育男性生殖组织中细胞凋亡失调的证据。在 目标1，生殖细胞在不同发育和分化阶段的凋亡脆弱性 使用功能性细胞死亡预测分析和睾丸辐射诱导生殖细胞死亡来确定。在 目标2，通过分析胚胎来研究细胞凋亡在男性性别分化中的要求 细胞凋亡不足遗传模型中的生殖道发育和成人 HPG 轴信号传导。 睾丸生殖细胞移植将用于确定生殖细胞与体细胞睾丸细胞的贡献 细胞凋亡失调导致男性不育。这些发现将支持确定针对性的治疗方法 影响发育中儿童的性发育障碍或细胞毒性暴露，导致贫困 成年后的生殖结果。 拟议研究金的培训计划将进一步发展分子和 发育生物学方法，并解决其他学科的概念和技术缺乏经验 生殖科学，包括内分泌学和流行病学。它还将提供从 将斑马鱼作为小鼠的疾病和发育模型。这将通过继续教育来实现 生物统计学，与共同赞助者一起进行培训，并利用跨学科科学咨询委员会进行监督 实验设计和数据分析。今年还将开展负责任的研究培训 1.哈佛大学公共卫生学院，致力于生殖健康和生殖健康的多方面研究 不孕不育症，将通过成像和分析设施以及 对该领域现有教师的监督。 Kristopher Sarosiek 的实验室擅长功能性 细胞凋亡的测定，将有助于开发一个独特而严格的研究计划，以桥接性 发育和细胞死亡的分子生物学，以揭示导致男性不育的机制。