Targeting sperm-specific proteins during meiosis and sperm morphogenesis

在减数分裂和精子形态发生过程中靶向精子特异性蛋白

• 批准号: 10164826
• 负责人: MARTIN M. MATZUK
• 金额: $ 27.86万
• 依托单位: BAYLOR COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-05-01 至 2024-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10164826
• 关键词: Acrosome; Affect; Alleles; Binding Proteins; Bioinformatics; Bromodomain; CRISPR/Cas technology; Chemistry; Child; Climate; Complex; Contraceptive Agents; Contraceptive methods; DNA; Development; Drug Targeting; Ecosystem; FDA approved; Fertility; Genes; Genomic DNA; Germ Cells; Goals; Hormones; Human; In Vitro; Integral Membrane Protein; Knock-out; Knockout Mice; Lead; Male Contraceptive Agents; Male Sterility; Meiosis; Membrane Proteins; Modeling; Morphogenesis; Mus; Mutant Strains Mice; Mutation; Names; Oral; Oral Contraceptives; Partner in relationship; Pharmaceutical Chemistry; Pharmaceutical Preparations; Phenotype; Phosphotransferases; Planets; Population; Population Growth; Proteins; Public Health; Recombinant Proteins; Resources; Scientist; Secondary to; Serine; Sperm Count Procedure; Sperm Motility; Spermatocytes; Spermatogenesis; Sterility; Technology; Testis; Water; Woman; X-Ray Crystallography; Zona Pellucida; base; contraceptive target; drug metabolism; druggable target; fly; food shortage; functional genomics; in vivo; inhibitor/antagonist; male; male fertility; men; metabolic abnormality assessment; multidisciplinary; novel; pill; pre-clinical; side effect; small molecule; small molecule inhibitor; sperm cell; sperm function; success

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.

项目 1 摘要（在减数分裂和精子形态发生过程中针对精子特异性蛋白） 项目1的总体目标是利用CRISPR/Cas9来了解精子的形成- 特定顶体并识别药物样探针和临床前候选物以靶向生精- 具有体内避孕作用的特定药物蛋白。人口数量已达到惊人水平 数字，导致我们的气候和生态系统发生变化，并可能导致全球食物、水和 为我们的子孙后代提供其他资源。遏制人口急剧增长及其可怕的后果 因此，我们需要专门针对男性生殖系的避孕药具。避孕药的目标 睾丸特异性蛋白质应该消除不需要的副作用。作为避孕目标，我们将重点关注两个方面 减数分裂特异性蛋白和四种精子必需的顶体相关跨膜蛋白。这六个 根据小鼠的原理验证研究选择了进化保守的蛋白质，表明 它们是男性生育能力所特别需要的，我们在提供小分子以实现可逆性方面取得了成功 体内避孕，以及这些蛋白质作为男性避孕药物靶标的潜力。七十 FDA 批准的药物中有 1% 是针对分泌蛋白或跨膜蛋白。此外，顶体如何 顶体形成过程中蛋白质的相互作用尚不清楚。定义相互作用组的形成和功能 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 这四个基因。为了确定小分子探针和临床前候选物，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 针对这些必需的生精蛋白，并创造出各种口服和植入式 男性和女性的避孕药具。项目1的具体目标是：1）利用CRISPR/Cas9功能化 了解顶体的形成以及四种精子必需跨膜蛋白的具体关系； 2) 采用 DEC-Tec 来识别小分子探针和临床前候选物以靶向生精- 特定的“可成药”蛋白质； 3) 评估小分子潜在的体内避孕作用 抑制剂。 P01 和项目 1 的成功依赖于项目 2 和 3 以及 DEC-Tec 的互动 核。 Ikawa 博士和他的同事创建了几个关键模型，并将生成 Aim 1 的标记等位基因。 Sonnenburg 博士和他的项目 3 团队将帮助为我们的 Aim 2 DEC-Tec 生产重组蛋白 屏幕。博士。 Young、Huang、Sonnenburg、Simmons、Yu、Lee 和 Li 是使用 DEC-Tec、药用药物的专家 用于发现先导化合物的化学、X 射线晶体学和药物代谢研究。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 解决这一紧迫问题并为男性和女性创造新颖且可逆的非激素避孕药。