Structure-function analysis of Mullerian Inhibiting Substance (MIS)

苗勒管抑制物质（MIS）的结构-功能分析

基本信息

批准号：
10471277
负责人：
THOMAS B THOMPSON
金额：
$ 38.93万
依托单位：
UNIVERSITY OF CINCINNATI
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-08-18 至 2026-05-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10471277
关键词：
Adopted Adult Affinity Agonist Anti-Mullerian Hormone Receptor Type II Binding Biochemical Biological Biological Assay Biological Markers Biological Models Biology Biophysics Cell Culture Techniques Cells Cervix Uteri Complex Crystallization Data Development Embryonic Development Engineering Extracellular Domain Family Female Fertility Fetus Future Generations Hand Human In Vitro Laboratories Ligand Binding Ligand Binding Domain Ligands Mammalian Oviducts Measures Membrane Proteins Modeling Molecular Mullerian-inhibiting substance receptor Mutation Nature Ovarian Ovarian Ablation Ovary Phosphotransferases Play Polycystic Ovary Syndrome Production Public Health Reagent Receptor Signaling Recombinant Proteins Reproducibility Reproduction Reproductive Biology Research Personnel Roentgen Rays Role Serum Sex Differentiation Signal Transduction Signaling Protein Specificity Structure Structure of paramesonephric duct Subcellular structure Surface Plasmon Resonance Syndrome TGFBR2 gene Testing Transforming Growth Factor beta Transforming Growth Factor beta Receptors Urogenital ridge structure Uterus Woman Work X-Ray Crystallography analog antagonist base bone morphogenetic protein receptor type I chemotherapy extracellular female reproductive system folliculogenesis granulosa cell human female in vivo insight male member mullerian-inhibiting hormone novel novel therapeutics ovarian reserve primary ovarian insufficiency rational design receptor reproductive reproductive system disorder small molecule structural biology substance use

项目摘要

Project Summary: Müllerian Inhibiting Substance (MIS) or Anti- Müllerian Hormone (AMH), originally identified for its role in male sex differentiation during development, has emerged as a significant molecule in female reproduction. For example, MIS plays an important role in regulating follicle development and serum levels and is now used as a measure of ovarian reserve. Mutations in MIS are associated with both male and female reproductive disorders, including Persistent Müllerian Duct Syndrome (PMDS) in males and Polycystic Ovary Syndrome (PCOS) in females. As a member of the TGFβ family, MIS signals through a type I and type II receptor. Uniquely, MIS signals through its own type II receptor MISRII and the type I receptor Alk2, which is utilized by multiple ligands. While previous studies have detailed TGFβ family ligand interactions, how MIS interacts, at the molecular level, with MISRII and Alk2 is unknown. The objective of this proposal is to understand how MIS interacts with its cognate receptors and how specific MIS signaling is generated through these receptors. Here, we will combine both structural and functional approaches, including X-ray crystallography, cell-based signaling assays, structure-based engineering, and biological models of fertility and ovarian protection. We will pursue the following three specific aims (1) determine the crystal structure of MIS with MISRII and Alk2, (2) characterize the MIS receptors and determine how the generate MIS-specific signaling, using both the extracellular and intercellular domains and (3) generate MIS analogs that will be tested in both in vitro binding and cell-based assays, along with in vivo assay including an AAV9 model of follicle suppression. Collectively, this proposal will uncover how MIS interacts with its receptors providing a platform for developing reagents that modify MIS activity with the potential for future application in reproductive therapies.

项目摘要：苗勒管抑制物质 (MIS) 或抗苗勒管激素 (AMH)，最初鉴定为由于其在发育过程中男性性别分化中的作用，已成为女性中的重要分子例如，MIS 在调节卵泡发育和血清水平方面发挥着重要作用。现在用作卵巢储备功能的衡量标准，MIS 突变与男性和女性均相关。生殖疾病，包括男性持续性苗勒氏管综合征 (PMDS) 和多囊卵巢女性综合症 (PCOS) 作为 TGFβ 家族的成员，MIS 通过 I 型和 II 型受体发出信号。独特的是，MIS 通过其自身的 II 型受体 MISRII 和 I 型受体 Alk2 发出信号，后者被虽然之前的研究详细介绍了 TGFβ 家族配体的相互作用，但 MIS 是如何相互作用的。分子水平上，MISRII 和 Alk2 是未知的。该提案的目的是了解 MIS 是如何实现的。与其同源受体相互作用以及如何通过这些受体产生特定的 MIS 信号传导。我们将结合结构和功能方法，包括 X 射线晶体学、基于细胞的信号传导我们将追求生育力和卵巢保护的分析、基于结构的工程以及生物模型。以下三个具体目标 (1) 使用 MISRII 和 Alk2 确定 MIS 的晶体结构，(2) 表征 MIS 受体并确定如何使用细胞外和细胞间结构域并 (3) 生成 MIS 类似物，将在体外结合和基于细胞的结合中进行测试试验，以及包括 AAV9 卵泡抑制模型在内的体内试验。揭示 MIS 如何与其受体相互作用，为开发改变 MIS 活性的试剂提供平台具有未来在生殖治疗中应用的潜力。