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: 6724469
• 负责人: Barry T. Hinton
• 金额: $ 22.51万
• 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/6724469
• 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，因此被认为是男性避孕药开发的主要目标。我们的策略是针对附睾内对男性生育能力至关重要的受体、激酶和磷酸酶。这些靶标是理想的，因为它们适合小分子量抑制剂，并且血液-附睾屏障不会成为此类抑制剂的巨大障碍。考虑到这些策略，我们的目标是检查孤儿酪氨酸激酶受体 c-Ros 作为男性避孕药开发的靶点。我们还有兴趣确定 c-Ros 上游调节因子的组成部分和下游靶标作为男性避孕药开发的靶标。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）验证调节c-Ros的表达和活性可以调节成年雄性小鼠的生育力的假设。为了实现这一特定目标，我们将在小鼠的初始部分中产生一种新型的条件性 c-Ros 敲除。 (2) 检验以下假设：c-Ros 的活性取决于其激酶结构域与其他结合蛋白（包括新型和已知的激酶和磷酸酶）的关联。与 c-Ros 活性相关的蛋白质将被视为开发男性避孕药的目标。我们将生成 SHP-1 磷酸酶的初始片段特异性条件敲除，该磷酸酶与 c-Ros 的激酶结构域结合。 (3) 检验以下假设：c-Ros 的活性取决于其胞外结构域与膜结合受体的关联。我们将产生代谢型谷氨酸受体样孤儿 G 蛋白偶联受体的初始片段特异性条件敲除，该受体是 c-Ros 的假定配体。在每只推定的不育基因敲除动物的初始片段中发生变化的下游激酶、磷酸酶和受体的表达和活性将被鉴定并进一步考虑作为避孕药开发的目标。因此，根据RFA的研究范围，我们将进行“对精子成熟过程的研究，目的是确定参与控制男性生育能力的具体目标”。