Protease regulation of ovarian recrudescence

卵巢复发的蛋白酶调节

• 批准号: 7760302
• 负责人: KELLY Ansley YOUNG
• 金额: $ 10.76万
• 依托单位: CALIFORNIA STATE UNIVERSITY LONG BEACH
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-01-01 至 2013-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7760302
• 关键词: Address; Atrophic; Breeding; CD34 gene; Cell Proliferation; Cleaved cell; Clinical; Contraceptive methods; Data; Development; Endocrine; Endopeptidases; Estradiol; Event; Exposure to; Extracellular Matrix; Family; Funding; Gelatinase A; Gelatinases; Goals; Gonadotropins; Growth and Development function; Hormonal; Hour; Hypothalamic structure; Individual; Inhibition of Matrix Metalloproteinases Pathway; Laboratories; Light; Link; Luteinizing Hormone; Matrix Metalloproteinase Inhibitor; Matrix Metalloproteinases; Measurement; Mediating; Messenger RNA; Metalloproteases; Methods; Modeling; Modification; Molecular; Organ; Ovarian; Ovarian Cycles; Ovarian Tissue; Ovary; Ovulation; Parents; Pathway interactions; Peptide Hydrolases; Periodicity; Phodopus sungorus; Photoperiod; Pituitary Gland; Plasma; Play; Premature Menopause; Process; Production; Progesterone; Prostaglandins; Proteins; Proteome; Proteomics; Protocols documentation; Recrudescences; Regulation; Reproduction; Reproductive Techniques; Research Design; Resources; Role; SB 3CT compound; Seasons; Series; Siberian Hamster; Steroid biosynthesis; Steroids; Testing; Tissues; Vascular Endothelial Growth Factors; Vascularization; angiogenesis; assisted reproduction; base; corpus luteum; cytokine; day length; folliculogenesis; functional status; gonad function; granulosa cell; in vivo; indexing; inhibitor/antagonist; insight; novel; offspring; public health relevance; reproductive; research study; response; restoration

DESCRIPTION (provided by applicant): Ovarian function, including follicle development, ovulation, and corpus luteum formation/degradation, is dependent upon tissue remodeling events, many of which are associated with a family of Zn+dependent endopeptidases, the matrix metalloproteinases (MMPs). Extreme remodeling of ovarian tissue is observed in photoperiodic species where seasonal changes in day length can either inhibit or stimulate hypothalamic/ pituitary secretion of GnRH/gonadotropins, leading to atrophy or resumption of ovarian function. Photo- responsive individuals are therefore excellent models for basic ovarian function, because modifications of day length naturally induce and then reverse ovarian atrophy. In photoperiodic Siberian hamsters, ovarian remodeling prompted by changes in day length is associated with differential expression of MMPs. Indeed, photostimulated return to ovarian function (recrudescence) can be impeded following in vivo administration of a broad-spectrum MMP inhibitor, GM6001. While this suggests an important role for these proteases, the action of MMPs during recrudescence and the process of how the quiescent ovary can resume cycling following weeks of atrophy is unknown. This proposal hypothesizes that 1) inhibition of recrudescence in response to GM6001 occurs because cleavage of MMP substrates that normally mediate return to ovarian function is impeded, 2) that the GM6001-treated ovary fails to return to function because key ovarian processes such as angiogenesis, granulosa cell proliferation, and steroidogenesis are dependent on MMP activity, and 3) that because gelatinases (MMPs-2/-9) are key players in ovarian cyclicity, and gelatinase activity is significantly down regulated following GM6001 administration, much of the remodeling can be attributed to gelatinases. Initial experiments will use a hypothesis-driven proteomics approach to identify substrates cleaved by MMPs during recrudescence by comparing the proteome of GM6001-treated vs. control ovaries. Examination of both mRNA and protein for key markers of angiogenesis (e.g., CD34, VEGF-R1), proliferation of granulosa cells (PCNA), and steroidogenesis (e.g., Cyp19, 32HSD) will reveal a mechanism of MMP action during recrudescence, and define a role for MMPs in these processes as GM6001 treated tissue is compared to controls. Finally, a gelatinase-specific inhibitor (SB-3CT) will be administered in vivo during photostimulated recrudescence to provide direct evidence of gelatinase action and function in the recovering ovary. Together, these studies should provide novel insight into the cellular and molecular regulation of recrudescence of ovarian function. In addition, these data will provide the first evidence of the targets of MMP action as ovarian cyclicity returns. Understanding the role that MMPs play in mammalian ovarian function by using a photoperiodic model helps to elucidate critical clinical questions of how to shut down (contraception) and restart (assisted reproduction, premature menopause) ovarian activity with non-hormonal mechanisms. PUBLIC HEALTH RELEVANCE: Ovarian recrudescence is a natural process stimulated by changes in photoperiod (the number of hours of light per day) whereby the atrophied and non-functional ovary returns to a fully functional organ in 2-8 weeks. While the hormonal initiation of recrudescence has been thoroughly examined, the cellular changes that occur at the level of the ovary remain unknown. Understanding the cellular/molecular pathways of mammalian ovarian recrudescence, such as matrix metalloproteinase (MMP) action, is novel and timely, as new non-hormonal methods of contraception and assisted reproduction techniques are clinically sought. Our studies will provide potential non-hormonal targets for clinically- geared studies focused on "turning on" and "turning off" ovarian function.

描述（由申请人提供）：卵巢功能，包括卵泡发育，排卵和叶黄素形成/降解，取决于组织重塑事件，其中许多与Zn+依赖性内肽酶家族有关，基督基质金属蛋白酶（MMPS）。在光周期物种中观察到卵巢组织的极端重塑，在这些物种中，白天长度的季节性变化可以抑制或刺激GNRH/促性腺激素的下丘脑/垂体分泌，从而导致卵巢萎缩或恢复卵巢功能。因此，响应式个体是基本卵巢功能的极好模型，因为白天长度的修饰自然诱导，然后逆向卵巢萎缩。在光周期的西伯利亚仓鼠中，白天长度变化引起的卵巢重塑与MMP的差异表达有关。实际上，在体内给药中宽光谱MMP抑制剂GM6001的体内给药后，光验证恢复到卵巢功能（复发）。尽管这表明这些蛋白酶的作用是重要的作用，但在复发过程中MMP的作用以及静脉内卵巢如何在萎缩周后如何恢复循环的过程。 This proposal hypothesizes that 1) inhibition of recrudescence in response to GM6001 occurs because cleavage of MMP substrates that normally mediate return to ovarian function is impeded, 2) that the GM6001-treated ovary fails to return to function because key ovarian processes such as angiogenesis, granulosa cell proliferation, and steroidogenesis are dependent on MMP activity, and 3）由于明胶酶（MMPS-2/-9）是卵巢循环的关键参与者，并且在GM6001给药后调节明胶活性显着下降，因此大部分重塑可以归因于明胶酶。最初的实验将使用假设驱动的蛋白质组学方法来鉴定MMP在复发过程中通过比较GM6001处理的蛋白质组与对照卵巢的蛋白质组来识别MMP裂解的底物。检查mRNA和蛋白质的血管生成主要标志物（例如CD34，VEGF-R1），颗粒细胞的增殖（PCNA）和类固醇生成（例如CYP19，32HSD）的检查将揭示MMP作用的机制，并在这些过程中与MMPS进行了iSS.611111的作用。最后，在光刺激复发期间，将在体内施用明胶酶特异性抑制剂（SB-3CT），以提供明胶酶作用和功能的直接证据。总之，这些研究应提供对卵巢功能复发的细胞和分子调节的新见解。此外，这些数据将为卵巢周期性回报提供第一个MMP作用目标的证据。了解MMP通过使用光周期模型在哺乳动物卵巢功能中扮演的作用有助于阐明如何通过非激素机制关闭（避孕）和重新启动（避孕）和重新启动（辅助繁殖，过早的更年期）卵巢活性的关键临床问题。 公共卫生相关性：卵巢复发是由光周期变化（每天的光小时数）刺激的自然过程，从而在2-8周内腐败和非功能性卵巢返回到功能齐全的器官。尽管已经彻底检查了复发的激素开始，但卵巢水平上发生的细胞变化仍然未知。在临床上寻求了新的非激素避孕技术和辅助复制技术，因此了解哺乳动物卵巢复发的细胞/分子途径，例如基质金属蛋白酶（MMP）作用是新颖且及时的。我们的研究将为临床上的研究提供潜在的非荷尔蒙靶标，该研究的重点是“打开”和“关闭”卵巢功能。