Epigenetics and Genetics of Infertility and Associated Comorbidities

项目总结 尽管对配子发生所需的基因有广泛的了解，但大多数人类不育的原因是 未知。据认为，大约有一半的病例有遗传基础，但要确定它仍然是有问题的。 确定确切的遗传损伤(S)，即使是基因组序列，也可能对给定的人负责 信息。鉴于有迹象表明某些基于基因的不孕症原因与 并存，以及人口中不孕不育的高发病率，这将对临床产生重大影响 在可靠地确定不孕不育的分子原因方面取得进展。非遗传原因的根源是 甚至更难确定，但表观遗传变化可能是某些类型不孕症的基础，特别是在 精原干细胞(SSCs)中可能发生这种变化的男性。项目II有3个具体目标 寻求解决这一领域的这些重要问题。目标1是从功能上验证候选基因 从项目1中的不孕症患者队列和家庭中识别出的变异。这将通过对 在CRISPR修饰的小鼠中进行变异，然后不仅对小鼠的生殖表型进行表型鉴定，而且还 相应患者可能存在的合并症，如肥胖、癌症易感性或 心血管疾病。目标2是表征导致不孕症或亚生育表型的突变。 小鼠，可能有尚未认识到的共病。重点将放在 除睾丸外，在体细胞组织中也有表达。AIM 3使用一种新的方法来识别表观遗传学 影响SSCs的改变，可能导致仅支持细胞综合征(SCOS)和非生殖 表型。这种方法涉及CRISPR抑制和激活技术，以进行 已知的影响体细胞重编程和SSC效率的表观遗传修饰基因 维修。总而言之，这个项目使用了多样化的尖端策略来提高准确性和发现能力 人类男性不育及相关并存的遗传和表观遗传原因。

PROJECT SUMMARY Despite extensive knowledge of genes required for gametogenesis, the causes of most human infertilities are unknown. It is thought that about half of cases have a genetic basis, but it remains problematic to definitively identify the exact genetic lesion(s) that may be responsible in a given person, even with genome sequence information. Given the indications that certain genetically-based causes of infertility are associated with comorbidities, and the high incidence of infertility in the population, it would be of substantial clinical impact to make progress towards reliably identifying molecular causes of infertility. The roots of non-genetic causes are even more difficult to ascertain, but epigenetic alterations may underlie certain types of infertility, particularly in males where such alterations may occur in spermatogonial stem cells (SSCs). Project II has 3 Specific Aims that seek to address these important issues in the field. Aim 1 is to functionally validate candidate genetic variants identified from infertility patient cohorts and families in Project 1. This will be done by modeling the variants in CRISPR-modified mice, then phenotyping the mice for not only reproductive phenotypes, but also comorbidities that may exist in the corresponding patients, such as obesity, cancer susceptibility, or cardiovascular disease. Aim 2 is to characterize mutations that cause infertility or subfertility phenotypes in mice, and which may have comorbidities that have yet to be recognized. An emphasis will be on genes that are expressed in somatic tissues in addition to the testis. Aim 3 employs a novel approach to identify epigenetic alterations that impact SSCs, potentially leading to Sertoli Cell Only syndrome (SCOS) and non-reproductive phenotypes. This approach involves CRISPR inhibition and activation technologies to conduct a screen of known epigenetic modification genes that impact the efficiency somatic cell reprogramming and SSC maintenance. In sum, this project uses diverse, cutting-edge strategies to improve the accuracy and discovery of genetic and epigenetic causes of human male infertility and related comorbidities.