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: 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
• 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

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 生殖。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 多个配体。虽然之前的研究详细介绍了 TGFβ 家族配体相互作用，但 MIS 是如何相互作用的， 分子水平上，MISRII 和 Alk2 未知。本提案的目的是了解 MIS 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 信号传导。这里， 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 分析、基于结构的工程以及生育和卵巢保护的生物模型。我们将追求 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 测定，以及体内测定，包括卵泡抑制的 AAV9 模型。总的来说，该提案将 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。