C-Ros pathways as targets for contraceptive development

C-Ros 途径作为避孕药开发的目标

• 批准号: 6831200
• 负责人: Barry T. Hinton
• 金额: $ 22.04万
• 依托单位: UNIVERSITY OF VIRGINIA
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-12-01 至 2008-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6831200
• 关键词: RNA interference; cell surface receptors; contraceptives; cooperative study; drug design /synthesis /production; electroporation; enzyme activity; epididymis; fertility; gene targeting; genetically modified animals; laboratory mouse; light microscopy; male; microinjections; phosphoprotein phosphatase; protein binding; protein tyrosine kinase; receptor expression; western blottings; yeast two hybrid system

DESCRIPTION (provided by applicant): The epididymis plays a crucial role in the maturation of spermatozoa and is, therefore, considered to be a prime target for the development of a male contraceptive. Our strategy is to target receptors, kinases and phosphatases within the epididymis that are crucial for male fertility. These targets are ideal because they are amenable to small molecular weight inhibitors and the blood-epididymis barrier would not be a formidable hurdle for such inhibitors. With these strategies in mind, our goal is to examine the orphan tyrosine kinase receptor c-Ros as a target for male contraceptive development. We are also interested in identifying the components of upstream regulators and downstream targets of c-Ros as targets for male contraceptive development. C-Ros was chosen because c-Ros knockout male mice are infertile. The specific aims of this proposal are: (1) To test the hypothesis that modulating the expression and activity of c-Ros can regulate fertility in adult male mice. To accomplish this specific aim we will generate a novel conditional c-Ros knockout in the initial segment of mice. (2) To test the hypothesis that the activity of c-Ros is dependent upon the association of its kinase domain with other binding proteins including novel and known kinases and phosphatases. Proteins that associate with c-Ros activity will be considered as targets for the development of a male contraceptive. We will generate an initial segment-specific conditional knockout of the SHP-1 phosphatase, which binds to the kinase domain of c-Ros. (3) To test the hypothesis that the activity of c-Ros is dependent upon its association of its extracellular domain with a membrane-bound receptor. We will generate an initial segment-specific conditional knockout of the metabotropic glutamate receptor-like orphan G-protein coupled receptor, which is the putative ligand for c-Ros. Expression and activity of downstream kinases, phosphatases and receptors that change in the initial segment of each putative infertile knockout animal will be identified and considered further as targets for contraceptive development. Therefore, in accordance with the research scope of the RFA, we will perform "research on the processes of sperm maturation with the goal of defining specific targets, involved in the control of male fertility."

描述（由申请人提供）：附睾在精子的成熟中起着至关重要的作用，因此被认为是发展男性避孕药的主要目标。我们的策略是在附睾中靶向受体，激酶和磷酸酶，这对于男性生育至关重要。这些靶标是理想的，因为它们适合小分子量抑制剂，而血液 - 抑制屏障对于此类抑制剂而言并不是巨大的障碍。考虑到这些策略，我们的目标是研究孤儿酪氨酸激酶受体C-ROS，作为男性避孕发育的目标。我们还有兴趣确定上游调节器和C-ROS下游目标的组成部分，作为男性避孕性发展的目标。之所以选择C-ROS，是因为C-ROS敲除雄性小鼠不育。该提案的具体目的是：（1）检验以下假设：调节C-ROS的表达和活性可以调节成年雄性小鼠的生育能力。为了实现这一特定目标，我们将在小鼠的初始段中产生一种新的条件C-ROS敲除。 （2）为了测试C-ROS活性取决于其激酶结构域与其他结合蛋白（包括新型和已知激酶和磷酸酶）的关联的假设。与C-ROS活性相关的蛋白质将被视为开发男性避孕药的靶标。我们将生成SHP-1磷酸酶的初始段特异性条件敲除，该磷酸酶与C-ROS的激酶结构域结合。 （3）测试C-ROS活性取决于其细胞外域与膜结合受体的关联的假设。我们将生成代谢型谷氨酸受体样孤儿G蛋白偶联受体的初始段特异性有条件敲除，这是C-ROS的假定配体。下游激酶，磷酸酶和受体在每个推定的不育敲除动物的初始段都会发生变化的表达和活性，并将进一步视为避孕发育的靶标。因此，根据RFA的研究范围，我们将“对精子成熟过程进行研究，目的是定义涉及男性生育能力的特定靶标”。