Structure-function analysis of Mullerian Inhibiting Substance (MIS)

苗勒管抑制物质（MIS）的结构-功能分析

• 批准号: 10280107
• 负责人: THOMAS B THOMPSON
• 金额: $ 41.99万
• 依托单位: UNIVERSITY OF CINCINNATI
• 依托单位国家: 美国
• 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/10280107
• 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; base; bone morphogenetic protein receptor type I; chemotherapy; design; 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; receptor; reproductive; reproductive system disorder; small molecule; structural biology; substance use

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.

项目概述：最初确定的苗勒氏抑制物质(MIS)或抗苗勒氏激素(AMH) 由于它在男性发育过程中的性别分化中的作用，它已经成为女性体内的一个重要分子 繁殖。例如，管理信息系统在调节卵泡发育和血清水平方面发挥着重要作用 现在被用作衡量卵巢储备的指标。男性和女性都与MI基因突变有关 生殖障碍，包括男性持续性苗勒管综合征(PMDS)和多囊卵巢 女性多囊卵巢综合征(PCOS)。作为转化生长因子β家族的一员，MISI通过I型和II型受体传递信号。 独一无二的是，管理信息系统通过它自己的II型受体MISRII和I型受体Alk2发出信号，后者被 多个配体。虽然之前的研究已经详细说明了转化生长因子β家族的配体相互作用，但在 分子水平上，与MISRII和ALK2是未知的。本提案的目标是了解管理信息系统是如何 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。这里, 我们将结合结构和功能方法，包括X射线结晶学、基于细胞的信号传递 生育力和卵巢保护的分析、基于结构的工程学和生物学模型。我们会继续推行 根据以下三个具体目标(1)用MISRII和ALK2确定了管理信息系统的晶体结构，(2)表征了 并确定如何使用细胞外和细胞外信号产生管理信息系统特异性信号 细胞间域和(3)产生将在体外结合和基于细胞的两者中进行测试的管理信息系统类似物 包括AAV9卵泡抑制模型在内的体内试验。总体而言，这项提议将 揭示管理信息系统如何与其受体相互作用，为开发修改管理信息系统活性的试剂提供平台 具有未来在生殖治疗中应用的潜力。