FUNCTIONAL GENOMICS AND PROTEOMICS TO REVEAL REPRODUCTIVE-TRACT SPECIFIC PROTEINS

摘要 这项提议的总体目标是通过高密度脂蛋白来阐明新的男性生殖道特异蛋白。 通过转录组学和蛋白质组学的方法，重点研究了五个新的基因分析 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， 需要有效和可逆的男性生育控制的非荷尔蒙方法，如现有的方法 对女性来说，是被广泛认可的。到目前为止，荷尔蒙男性避孕药还没有效果。因此， 在精子和男性生殖道中识别新的可用药靶点需要取得进展 男性避孕药的开发。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，尽管从 小鼠和大鼠附睾节段，很少有已发表的关于人类附睾节的数据集。随着精子的进步 沿着附睾管，它们沐浴在特殊的管腔液体微环境中，这对 他们的成熟和生存。考虑到它的重要作用，附睾会成为 男性避孕药的发展。本研究将为以后的工作奠定基础。 针对新目标的男性避孕药的开发。活泼、成熟的精子是成功的关键 受精和我们的总体假设是在人类男性中存在未发现的可用药靶点 对精子成熟和功能至关重要的生殖道。我们的具体目标如下：1) 对人类附睾节进行转录分析，以确定新的附睾特异性药物 干扰精子成熟的靶点；2)对人类精子进行蛋白质组分析以鉴定新的 精子特异的避孕药物靶点；3)在小鼠身上应用CRISPR/Cas9来验证关键精子 或精子功能和/或受精所需的生殖道特异性蛋白。该计划的最终目标 拟议的工作是导致确定更多的新目标，一个跨学科团队 将致力于。我们的机制研究将使我们能够将这些蛋白质放入生殖途径中，并 同时确定每种蛋白质作为避孕靶标的效用。

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.