Targeting sperm-specific proteins during meiosis and sperm morphogenesis

项目1总结(针对减数分裂和精子形态发生过程中的精子特定蛋白) 项目1的总体目标是使用CRISPR/Cas9来了解精子的形成- 特定的顶体和识别药物样探针和临床前候选对象来靶向生精- 体内具有避孕效果的特定可药物蛋白质。人口数量正在达到令人震惊的水平 数字，导致我们的气候和生态系统的变化，以及可能的全球食物、水和 为我们的子孙提供的其他资源。遏制猖獗的人口增长及其可怕的局面 后果，我们要求专门针对男性生殖系的避孕药具。针对目标的避孕药 睾丸特异的蛋白质应该会消除不必要的副作用。作为避孕目标，我们将重点关注两个方面 减数分裂特异蛋白和四种精子必需的顶体相关跨膜蛋白。这六个人 基于对小鼠的原理验证研究，选择了进化上保守的蛋白质 它们是男性生育所必需的，我们成功地提供了可逆的小分子 体内避孕，以及这些蛋白质作为男性避孕药物靶点的潜力。七十 FDA批准的药物中有1%针对分泌型或跨膜蛋白。此外，顶体是如何 蛋白质在顶体形成过程中的相互作用尚不清楚。为…的形成和功能定义互动体 顶体，我们将与Ikawa博士(项目2)合作，使用CRISPR/Cas9将标签序列插入到 这四个基因。为了识别小分子探针和临床前候选分子，我们将与DNA- 编码化学技术(DEC-Tec)核心。我们的总体假设是CRISPR/CAS9和DEC-Tec 将帮助我们定义精子发生过程中的蛋白质相互作用组，快速识别多个小分子 它们针对这些基本的生精蛋白，并创造出各种口服和植入的 男性和女性的避孕药。项目1的具体目标是：1)在功能上使用CRISPR/CAS9 了解顶体形成和四种精子必需跨膜蛋白的特殊关系； 2)使用DEC-Tec来识别小分子探针和临床前候选对象，以靶向生精- 3)评价小分子药物的体内潜在避孕效果。 抑制剂。P01和项目1的成功有赖于与项目2和项目3以及DEC-Tec的互动 核心。池川博士和他的同事们创建了几个关键模型，并将为AIM 1生成标记的等位基因。 Sonnenburg博士和他的Project 3团队将帮助为我们的Aim 2 DEC-Tec生产重组蛋白质 屏幕。Young，Huang，Sonnenburg，Simmons，Yu，Lee和Li博士是使用DEC-Tec，Medical 化学、X射线结晶学和药物代谢研究，以发现铅化合物。项目1将 对所有三个项目进行体内概念验证研究(目标3)。因此，我们的多学科团队将 解决这一紧迫问题，为男性和女性创造新的、可逆的非激素避孕药。

PROJECT 1 SUMMARY (Targeting sperm-specific proteins during meiosis and sperm morphogenesis) The overall goals of Project 1 are to use CRISPR/Cas9 to understand the formation of the sperm- specific acrosome and identify drug-like probes and preclinical candidates to target spermatogenic- specific druggable proteins for a contraceptive effect in vivo. The human population is reaching alarming numbers, resulting in changes in our climates and ecosystems and likely global shortages of food, water, and other resources for our children and grandchildren. To curb rampant population growth and its dire consequences, we require contraceptives that specifically target the male germline. Contraceptives that target testis-specific proteins should eliminate unwanted side effects. As contraceptive targets, we will focus on two meiosis-specific proteins and four sperm-essential, acrosome-associated transmembrane proteins. These six evolutionarily-conserved proteins were chosen based on proof-of-principle studies in the mouse showing that they are specifically required for fertility in males, our success at delivering small molecules for reversible contraception in vivo, and the potential of these proteins as druggable targets for male contraception. Seventy percent of FDA-approved drugs target either secreted or transmembrane proteins. Further, how the acrosomal proteins interact during acrosome formation is unclear. To define the interactome for formation and function of the acrosome, we will collaborate with Dr. Ikawa (Project 2) to use CRISPR/Cas9 to insert tag sequences into these four genes. To identify small-molecule probes and preclinical candidates, we will partner with the DNA- Encoded Chemistry Technology (DEC-Tec) Core. Our overall hypothesis is that CRISPR/Cas9 and DEC-Tec will help us to define the protein interactome during spermatogenesis, rapidly identify multiple small-molecules that are directed at these essential spermatogenic proteins, and create an assortment of oral and implantable contraceptives for men and women. The Specific Aims of Project 1 are: 1) Use CRISPR/Cas9 to functionally understand acrosome formation and the specific relationship of four sperm-essential transmembrane proteins; 2) Employ DEC-Tec to identify small-molecule probes and preclinical candidates to target spermatogenic- specific “druggable” proteins; and 3) Evaluate potential in vivo contraceptive effects of small-molecule inhibitors. The success of this P01 and Project 1 relies on interactions with Projects 2 and 3 and the DEC-Tec Core. Dr. Ikawa and his colleagues created several key models and will generate the tagged alleles for Aim 1. Dr. Sonnenburg and his Project 3 team will help to produce recombinant proteins for our Aim 2 DEC-Tec screens. Drs. Young, Huang, Sonnenburg, Simmons, Yu, Lee, and Li are experts at using DEC-Tec, medicinal chemistry, X-ray crystallography, and drug metabolism studies for uncovering lead compounds. Project 1 will perform the in vivo proof-of-concept studies for all three projects (Aim 3). Thus, our multidisciplinary team will tackle this urgent problem and create novel and reversible non-hormonal contraceptives for men and women.