Transforming growth factor β family signaling pathways in ovarian and uterine biology

卵巢和子宫生物学中转化生长因子β家族信号通路

• 批准号: 10611376
• 负责人: MARTIN M. MATZUK
• 金额: $ 47.56万
• 依托单位: BAYLOR COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 1994
• 资助国家: 美国
• 起止时间: 1994-08-17 至 2025-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10611376
• 关键词: Acceleration; Activin A type II receptor; Activin Receptor; Activins; Address; Advanced Development; Affinity; Award; BMP15 gene; Binding; Binding Proteins; Biochemical; Biology; CRISPR/Cas technology; Cachexia; Cell Nucleus; Cell secretion; Cells; Chemicals; Chemistry; Clinic; Clinical Treatment; Complex; Contraceptive Agents; Contraceptive methods; DNA; DNA Sequence; Defect; Development; Diagnosis; Disease; Endometrial Hyperplasia; Event; Family; Family member; Female; Fertility; Fetal Growth Retardation; Follistatin; Functional disorder; Funding; GDF11 gene; GDF8 gene; Genes; Genetic; Genetic Transcription; Genomics; Germ Cells; Goals; Grant; Growth Differentiation Factor 9; Human; INHA gene; Infertility; Inhibin A; Investigation; Knock-out; Knockout Mice; Libraries; Ligands; Link; MADH2 gene; MADH3 gene; MADH4 gene; Malignant neoplasm of ovary; Mammals; Mediating; Medicine; Mission; Molecular; Molecular Mechanisms of Action; Mus; Muscle; Muscular Atrophy; National Institute of Child Health and Human Development; Nature; Oocytes; Ovarian; Ovary; Paper; Pathway interactions; Pharmaceutical Preparations; Phosphorylation; Phosphotransferases; Physicians; Physiological; Physiology; Pituitary Gland; Play; Population Control; Pre-Eclampsia; Pregnancy; Pregnancy loss; Procedures; Process; Productivity; Protein Secretion; Proteomics; Publishing; Pulmonary Hypertension; Quality of life; Receptor Serine/Threonine Kinase; Receptor Signaling; Recurrence; Regulation; Reproduction; Reproductive Health; Research; Role; Science; Scientist; Secondary to; Signal Pathway; Signal Transduction; Smad Proteins; Somatic Cell; Specificity; Study models; Syndrome; TGF-beta type I receptor; Technology; Time; Transforming Growth Factor beta; Translational Research; Tumor Suppressor Proteins; Uterus; Woman; Women' s Health; antagonist; bone loss; bone morphogenetic protein receptors; cell type; dimer; endometriosis; female fertility; field study; folliculogenesis; follow-up; granulosa cell; implantation; improved; in vivo; inhibin; inhibitor; innovation; insight; kinase inhibitor; mouse genetics; mouse genome; mouse model; novel; novel therapeutics; offspring; protein function; receptor; reproductive; reproductive system disorder; reproductive tract; secondary infertility; small molecule; small molecule inhibitor; small molecule libraries; tool

PROJECT SUMMARY The transforming growth factor β (TGFβ) superfamily is the largest family of secreted proteins in mammals. These dimeric ligands, which function in nearly every developmental, physiologic, and pathophysiologic process, including infertility, signal through a heterodimeric complex of type 2 and type 1 serine-threonine kinase receptors that phosphorylate downstream regulatory SMAD proteins and bind SMAD4 to regulate transcription. With NICHD support that started with a physician scientists award (K11HD00960; 1991-93) and this R01 grant (1994-present), we have been productive leaders in the identification and characterization of the oocyte-secreted TGFβ family members, growth differentiation factor 9 (GDF9) and bone morphogenetic protein 15 (BMP15), and granulosa cell-secreted activins and inhibins. We have published extensively in this field including >30 papers in Nature, Nature Genetics, Nature Medicine, PLoS Biology, PLoS Genetics, PNAS, and Science. Whereas mammalian oocytes were initially hypothesized to be passengers rather than drivers in ovarian folliculogenesis, we showed that GDF9 is essential for fertility, discovered the X-linked BMP15 gene, and showed that GDF9:BMP heterodimers are the most active oocyte-secreted ligand in mice and women. These insights have defined the oocyte-somatic cell dialogue in ovarian folliculogenesis. In parallel, we showed that inhibin α-knockout mice are infertile, develop ovarian cancers and die due to an activin-induced cachexia syndrome. BMPs, GDF9:BMP15, activins, and myostatin share common type 2 receptors [activin receptor type 2A (ACVR2A) or type 2B (ACVR2B) or BMPR2], type 1 receptors (ALK4 and ALK5), and receptor-regulated SMADs (SMAD1,2,3,5). Using mouse genetics, we have shown that these proteins function in the pituitary, ovaries, and uterus (e.g., granulosa-specific knockout of SMAD2 and SMAD3 leads to cumulus defects and infertility due to defective GDF9:BMP15 signaling, whereas uterine-specific knockout of SMAD2 and SMAD3 leads to infertility secondary to endometrial hyperplasia). Although SMAD2 and SMAD3 play redundant roles in GDF9:BMP15, activin, and TGFβ signaling, we know little about the transcriptional complexes or DNA sequences that they bind. In addition, there are no small molecule inhibitors of ACVR2A/2B and BMPR2. Our overall hypothesis is that oocyte GDF9:BMP15, granulosa cell activins, and uterine BMPs and TGFβs signal through unique SMAD-mediated transcriptional complexes to regulate ovarian and uterine physiology in mice and women. Our proposal will take advantage of state-of-the-art CRISPR/Cas9 strategies to manipulate the mouse genome and DNA-encoded chemical libraries to create novel inhibitors of ACVR2A/2B and BMPR2 and perform follow-up genetic, proteomic, and biochemical approaches to reach our goals. At the end of 5 years, we expect to have unlocked key molecular events that are orchestrated by TGFβ family ligands in the female reproductive tract, thereby accelerating translational research to optimize assisted reproductive procedures for women and to create the first ACVR2A/2B and BMPR2 specific small molecules to regulate female fertility.

项目概要 转化生长因子 β (TGFβ) 超家族是哺乳动物中最大的分泌蛋白家族。 这些二聚体配体几乎在所有发育、生理和病理生理学中发挥作用 过程，包括不孕症，通过 2 型和 1 型丝氨酸-苏氨酸异二聚体复合物发出信号 磷酸化下游 SMAD 调节蛋白并结合 SMAD4 进行调节的激酶受体 NICHD 的支持始于医师科学家奖（K11HD00960；1991-93）和 在这项 R01 资助（1994 年至今）中，我们在识别和描述 卵母细胞分泌的 TGFβ 家族成员、生长分化因子 9 (GDF9) 和骨形态发生蛋白 15 (BMP15)，以及颗粒细胞分泌的激活素和抑制剂，我们主要在这个领域发表文章。 包括超过 30 篇发表在 Nature、Nature Genetics、Nature Medicine、PLoS Biology、PLoS Genetics、PNAS 和 科学。 哺乳动物卵母细胞最初被限制为乘客而不是司机。 卵巢卵泡发生，我们证明 GDF9 对于生育至关重要，发现了 X 连锁 BMP15 基因， 并表明 GDF9:BMP 异二聚体是小鼠和女性中最活跃的卵母细胞分泌配体。 这些见解同时定义了卵巢卵泡发生中的卵母细胞-体细胞对话。 抑制素 α 基因敲除小鼠不育，发展为卵巢癌，并因激活素诱导的恶病质而死亡 BMP、GDF9:BMP15、激活素和肌肉生长抑制素具有共同的 2 型受体 [激活素受体类型]。 2A (ACVR2A) 或 2B 型 (ACVR2B) 或 BMPR2]、1 型受体（ALK4 和 ALK5）以及受体调节型 SMAD (SMAD1,2,3,5) 使用小鼠遗传学，我们已经证明这些蛋白质在垂体中发挥作用， 卵巢和子宫（例如，SMAD2 和 SMAD3 的颗粒特异性敲除会导致卵丘缺陷和 GDF9:BMP15 信号传导缺陷导致不孕，而子宫特异性敲除 SMAD2 和 SMAD3 尽管 SMAD2 和 SMAD3 在其中发挥着冗余作用，但会导致继发于子宫内膜增生的不孕。 GDF9：BMP15、激活素和 TGFβ 信号传导，我们对转录复合物或 DNA 知之甚少 此外，我们的 ACVR2A/2B 和 BMPR2 没有小分子抑制剂。 总体假设是卵母细胞 GDF9:BMP15、颗粒细胞激活素、子宫 BMP 和 TGFβ 信号 通过独特的 SMAD 介导的转录复合物来调节小鼠卵巢和子宫生理机能 我们的建议将利用最先进的 CRISPR/Cas9 策略来操纵 小鼠基因组和 DNA 编码的化学文库，用于创建 ACVR2A/2B 和 BMPR2 的新型抑制剂 执行后续遗传、蛋白质组学和生化方法以实现我们的目标 5 年后， 我们期望解开女性中由 TGFβ 家族配体精心策划的关键分子事件 生殖道，从而加速转化研究，优化辅助生殖程序 女性并创造了第一个 ACVR2A/2B 和 BMPR2 特异性小分子来调节女性生育能力。