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.

项目摘要/摘要 在整个物种中，性发育和生育能力取决于细胞死亡事件。在发展中 哺乳动物的卵巢和睾丸，生殖细胞的大量死亡（精子和卵的前体）发生在 胎儿流失事件被认为是为了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回归过程中，在男性中也观察到 垂体的形态发生，可调节性腺性激素的产生。令人惊讶的是，抑制作用 凋亡导致小鼠男性特异性不育症，但尚不理解为什么会发生或细胞死亡 在整个男性性发展中受到监管。我的研究的目的是了解 沿着男性性发育中的下丘脑 - 垂体 - 基达（HPG）轴凋亡，以及为什么 凋亡的破坏会导致不育。具体而言，该建议旨在绘制正常的凋亡灵敏度 睾丸生殖细胞，并找到不育男性生殖组织中凋亡的疾病的证据。在 AIM 1，在不同的发育和分化阶段，生殖细胞的凋亡脆弱性将是 使用功能性细胞死亡预测性测定和睾丸辐照来诱导生殖细胞死亡。在 AIM 2，将通过分析胚胎来研究男性性别差异化凋亡的要求 凋亡功能不全的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建立种系与体细胞细胞的贡献 对男性不育症的凋亡失调。这些发现将支持确定针对性治疗的 性发育的疾病或影响发展中儿童的细胞毒性暴露，这导致贫困 成年人的生殖结果。 拟议奖学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不经验 生殖科学，包括内分泌学和流行病学。它还将提供从 斑马鱼对小鼠作为疾病和发育模型。这将通过继续教育 生物统计学，与共同发起人的培训，并利用跨学科的科学咨询委员会来监督 实验设计和数据分析。负责任的研究培训也将在一年中进行 1。哈佛公共卫生学院，致力于多方面的生殖健康研究 不育，将通过成像和分析设施以及 监督该领域已建立的教师。克里斯托弗·萨罗西克（Kristopher Sarosiek）的实验室，在功能上擅长 细胞凋亡的测定将使能够开发一个独特而严格的研究计划，以弥合性 开发和细胞死亡的分子生物学，以发现驱动男性不育症的机制。