FUNCTIONAL GENOMICS AND PROTEOMICS TO REVEAL REPRODUCTIVE-TRACT SPECIFIC PROTEINS

总结 这项计划的总体目标是通过高表达的蛋白质来阐明新的雄性生殖道特异性蛋白质。 通量转录组学和蛋白质组学方法，并专注于五个新的基因分析 表达仅限于男性生殖道的基因。随着世界人口继续急剧增长， 需要有效和可逆的非激素男性生育控制方法，如现有的方法， 对女性来说，这是公认的。到目前为止，荷尔蒙男性避孕药还没有效果。因此，我们认为， 需要在精子和男性生殖道中鉴定新的可药用靶点， 男性避孕药的发展。尽管蛋白质组学取得了重大进展，但一个全面的人类 精子蛋白质组尚未建立。同样地，尽管大量的转录组学数据发表于 小鼠和大鼠附睾段，很少有关于人类附睾的公开数据集。随着精子的发育 沿着附睾管，它们沐浴在一个特殊的管腔流体微环境中， 他们的成熟和生存。考虑到它的重要作用，附睾是治疗的主要目标。 开发男性避孕药具。预计拟议的研究将为未来的研究奠定基础。 开发针对新目标的男性避孕药。活跃成熟的精子是成功的关键 我们的总体假设是，人类男性中存在未发现的药物靶点， 生殖道对精子的成熟和功能至关重要。我们的具体目标如下：1） 对人类附睾段进行转录组学分析以鉴定新型附睾特异性药物 干扰精子成熟的靶点; 2）对人类精子进行蛋白质组学分析，以鉴定新的 用于避孕的精子特异性药物靶点;以及3）在小鼠中应用CRISPR/Cas9以验证关键精子 或精子功能和/或受精所需的生殖道特异性蛋白质。的最终目标 建议的工作是导致确定额外的新目标，一个跨学科的团队 将致力于。我们的机制研究将使我们能够将这些蛋白质置于生殖途径中， 同时确定这些蛋白质中的每一种作为避孕靶点的效用。

SUMMARY The overall goals of this proposal are to elucidate novel male reproductive tract-specific proteins through high throughput transcriptomics and proteomics approaches, and to focus on the genetic analyses of five novel genes with expression limited to the male reproductive tract. With the world population continuing its steep rise, the need for effective and reversible non-hormonal methods of fertility control for men, such as those available to women, is widely recognized. To date, hormonal male contraceptives have not been effective. Therefore, identification of novel druggable targets in sperm and the male reproductive-tract are required to advance development of male contraceptives. Despite significant advances in proteomics, a comprehensive human sperm proteome has not been established. Likewise, despite extensive transcriptomic data published from mouse and rat epididymal segments, few published datasets exist for human epididymis. As sperm progress along the epididymal duct, they are bathed in a specialized luminal fluid microenvironment that is crucial for their maturation and survival. Considering its essential role, the epididymis is a prime target for the development of contraceptives for men. The proposed research is anticipated to lay the groundwork for future development of male contraceptives against novel targets. Motile, mature sperm are essential for successful fertilization and our overall hypothesis is that undiscovered druggable targets exist in the human male reproductive tract that are critical for sperm maturation and function. Our specific aims are as follows: 1) Perform transcriptomic analysis of human epididymal segments to identify novel epididymis-specific druggable targets to interfere with sperm maturation; 2) Perform proteomic analysis of human sperm to identify novel sperm-specific druggable targets for contraception; and 3) Apply CRISPR/Cas9 in mice to validate key sperm or reproductive tract-specific proteins required for sperm function and/or fertilization. The end goal of the proposed to work is to lead to the identification of additional novel targets for which an interdisciplinary team will work toward. Our mechanistic studies will allow us to place these proteins into reproductive pathways and simultaneously determine the utility of each of these proteins as a contraceptive target.