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.

项目总结 转化生长因子β超家族是哺乳动物中最大的分泌型蛋白家族。 这些二聚体配体在几乎所有的发育、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-苏氨酸的异二聚体复合体发出信号 使下游调节蛋白Smad4磷酸化并结合Smad4调节的激酶受体 抄写。NICHD的支持始于医生科学家奖(K11HD00960；1991-93)和 这笔R01赠款(1994年至今)，我们一直是识别和表征 卵母细胞分泌的转化生长因子β家族成员、生长分化因子9和骨形态发生蛋白 15(BMP15)和颗粒细胞分泌的激活素和抑制素。我们在这一领域发表了大量的文章。 收录《自然》、《自然遗传学》、《自然医学》、《PLoS生物学》、《PLoS遗传学》、《美国国家科学院院刊》等30篇论文。 科学。而哺乳动物卵母细胞最初被假设为乘客而不是司机 我们发现了X连锁的BMP15基因，表明GDF9对生育是必不可少的。 研究表明，GDF9：BMP异源二聚体是小鼠和女性卵母细胞分泌最活跃的配体。 这些见解定义了卵泡发生中的卵母细胞-体细胞对话。与此同时，我们展示了 抑制素α基因敲除的小鼠不育，患上卵巢癌，并死于激活素诱导的恶病质 综合症。BMPS、GDF9：BMP15、激活素和肌肉生长抑素共享共同的2型受体[激活素受体类型 2A(ACVR2A)或2B(ACVR2B)或BMPR2]，1型受体(ALK4和ALK5)，以及受体调节的 SMADS(SMAD1、2、3、5)利用小鼠遗传学，我们已经证明了这些蛋白质在脑下垂体中起作用， 卵巢和子宫(例如，颗粒特异性Smad2和Smad3的敲除会导致卵丘缺陷和 GDF9：BMP15信号缺陷导致的不孕不育，而子宫特异的Smad2和Smad3基因敲除 导致继发于子宫内膜增生症的不孕)。尽管Smad2和Smad3在 骨形态发生蛋白15、激活素和转化生长因子β信号，我们对转录复合体或dna知之甚少。 它们结合的序列。此外，也没有ACVR2A/2B和BMPR2的小分子抑制剂。我们的 总体假设为卵母细胞GDF9：BMP15、颗粒细胞激活素，以及子宫BMPs和转化生长因子βS信号 通过独特的SMAD介导的转录复合体调节小鼠卵巢和子宫的生理 还有女人。我们的提案将利用最先进的CRISPR/Cas9战略来操纵 小鼠基因组和DNA编码的化学文库，以创建ACVR2A/2B和BMPR2的新抑制剂和 进行后续的遗传学、蛋白质组学和生化方法，以达到我们的目标。在五年结束的时候， 我们期望在女性体内解锁由转化生长因子β家族配体调控的关键分子事件。 生殖道，从而加速翻译研究，以优化辅助生殖程序 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。