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

基本信息

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
项目类别：
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

项目摘要

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靶向精子特异性蛋白）项目1的总体目标是使用CRISPR/CAS9了解精子的形成。特定的Adrosom体并确定类似药物的问题和临床前候选者，以靶向精子 - 特异性药物蛋白质可在体内产生避孕作用。人口令人震惊数字，导致我们的气候和生态系统的变化，以及全球食物，水和可能的短缺我们的孩子和孙子的其他资源。遏制人口猖ramp的增长及其可怕后果，我们需要专门针对男性种系的避孕药。针对目标的避孕药睾丸特异性蛋白质应消除不良的副作用。作为避孕目标，我们将专注于两个减数分裂特异性蛋白质和四个精子的，与邻域相关的跨膜蛋白。这六个根据小鼠的原理研究选择进化保存的蛋白质，表明该蛋白它们是在男性中专门需要的，我们成功地传递了小分子以进行可逆的分子体内避孕，这些蛋白质作为雄性避孕的可药靶标的潜力。七十 FDA批准的药物的百分比靶向分泌或跨膜蛋白。此外，如何访问蛋白质在高质体形成过程中相互作用尚不清楚。定义互动组以形成和功能 Acrosos，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。为了确定小分子问题和临床前候选者，我们将与DNA合作编码化学技术（DEC-TEC）核心。我们的总体假设是CRISPR/CAS9和DEC-TEC 将帮助我们在精子发生过程中定义蛋白质相互作用，并迅速识别多种小分子针对这些必需的精子蛋白，并创建各种口服和植入男女避孕药具。项目1的具体目的是：1）使用CRISPR/CAS9在功能上在形成之间了解四个精子跨膜蛋白的特定关系； 2）采用DEC-TEC识别小分子问题和临床前候选者，以靶向精子特定的“可药”蛋白； 3）评估小分子的体内避孕作用抑制剂。该P01和项目1的成功依赖于与项目2和3的互动以及DEC-TEC 核。 Ikawa博士和他的同事创建了多个关键模型，并将为AIM 1生成标签等位基因。 Sonnenburg博士和他的项目3团队将帮助我们为我们的目标生产重组蛋白2 dec-tec 屏幕。博士。 Young，Huang，Sonnenburg，Simmons，Yu，Lee和Li是使用DEC-TEC的专家用于发现铅化合物的化学，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（AIM 3）。那，我们的多学科团队将解决这个紧急问题，并为男性和女人创建新颖而可逆的非荷尔蒙避孕药。