Corpus luteum mechanical regulation in a tissue-engineered model of ovarian aging

卵巢衰老组织工程模型中黄体的机械调节

• 批准号: 8203279
• 负责人: Robin M. Skory
• 金额: $ 4.68万
• 依托单位: NORTHWESTERN UNIVERSITY AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-07-06 至 2015-07-05
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8203279
• 关键词: Actins; Address; Affect; Age; Age-Years; Aging; Alginates; Animals; Applications Grants; Biomedical Engineering; Cardiovascular Diseases; Cellular Structures; Characteristics; Clinical Trials; Comorbidity; Cytoskeleton; Diagnosis; Environment; Extracellular Matrix; Female; Fertility; Fibrosis; Figs - dietary; Functional disorder; Gland; Goals; Heart Diseases; Hormonal Change; Hormones; Hydrogels; In Vitro; Infertility; Integrin Binding; Integrins; Lead; Link; Longitudinal Studies; Luteal Cells; Luteal Phase; Mechanical Stress; Mechanics; Mediating; Menopause; Menstrual cycle; Modeling; Molecular; Mus; Organ; Osteoporosis; Ovarian; Ovarian hormone; Ovary; Pathway interactions; Phase; Phenocopy; Physical environment; Play; Premenopause; Process; Production; RGD (sequence); Regulation; Role; Signal Pathway; Signal Transduction; Steroid biosynthesis; Stress Fibers; Structure; System; Testing; Tissue Engineering; Tissues; Woman; Women' s Health; age related; base; cardiovascular disorder risk; cell type; cohort; corpus luteum; density; improved; in vivo; inhibitor/antagonist; juvenile animal; mouse model; novel; older women; proliferative phase Menstrual cycle; receptor binding; reproductive; reproductive axis; reproductive hormone; research study; response; rho; rho GTP-Binding Proteins; theories

DESCRIPTION (provided by applicant): Ovarian aging permanently alters the female hormone profile, causing sequellae such as infertility, osteoporosis, and cardiovascular disease. Before menopause, age-related changes in hormone production occur during both phases of the menstrual cycle. When compared to younger counterparts, older women have lower levels of luteal phase hormones, which are produced by the corpus luteum (CL). Unfortunately, the mechanism of age-related CL dysfunction is unknown. Ovarian aging is a fibrotic process involving dramatic stromal remodeling, which increases the microenvironment rigidity. While it is known that ECM- derived signals regulate CL steroidogenesis, the causative link between microenvironment rigidity and hormone production has not previously been shown. Thus, in the studies proposed herein, we will test the hypothesis that changes in the ovarian physical environment cause progressive luteal phase hormone decline in an aging mouse model. In preliminary studies, we have cultured murine CLs in a novel 3D alginate system, which can be tuned to different mechanical rigidities. In Aim 1, proposed experiments will manipulate alginate rigidity and define the effects on CL structure and steroidogenesis. Specifically, experiments will phenocopy older CLs by culturing glands from younger animals in more rigid alginate conditions. Conversely, a rescue experiment will be performed with CLs from older animals cultured in less rigid alginate environments. In addition, the relationship of ECM adhesivity and mechanical force will be explored by competitively inhibiting integrin binding with free RGD peptides. In the second Aim of this proposal, experiments will test a possible mechanism of luteal cell mechanotransduction. In many cell types, Rho/ROCK signaling plays a chief role in sensing the physical milieu and is a known effector of ECM-integrin engagement. Moreover, Rho/ROCK signaling could be the critical link between ECM rigidity, integrin binding, and luteal cell structure-function. Experiments will assess Rho/ROCK signaling in younger and older animal cohorts. Additionally, we will test Rho/ROCK signaling in CLs cultured in various rigidities using our 3D hydrogel system and several pathway inhibitors. By harnessing the ability to bioengineer the ovarian microenvironment, we can identify regulatory mechanisms at the intersection of age, tissue rigidity, and hormone production. In so doing, we will test a novel theory of female reproductive aging and identify mechanisms that may be regulated in vivo to improve luteal function in the aging, premenopausal women. PUBLIC HEALTH RELEVANCE: Changes in ovarian hormones play an important role in women's health as they age, increasing the risk for cardiovascular disease, osteoporosis and infertility. The goal of this project is to understand how age-related ovarian fibrosis alters hormone production. Determining the consequences of ovarian fibrosis will improve the diagnosis and treatment of hormone-related problems in aging, premenopausal women.

描述（由申请人提供）：卵巢衰老会永久改变女性荷尔蒙分布，导致不孕、骨质疏松和心血管疾病等后遗症。绝经前，月经周期的两个阶段都会发生与年龄相关的激素产生变化。与年轻女性相比，老年女性的黄体期激素水平较低，黄体期激素是由黄体 (CL) 产生的。不幸的是，与年龄相关的 CL 功能障碍的机制尚不清楚。卵巢衰老是一个涉及剧烈基质重塑的纤维化过程，这会增加微环境的刚性。虽然众所周知 ECM 衍生信号可调节 CL 类固醇生成，但微环境刚性与激素产生之间的因果关系此前尚未被证实。因此，在本文提出的研究中，我们将检验以下假设：在衰老小鼠模型中，卵巢物理环境的变化导致黄体期激素进行性下降。在初步研究中，我们在新型 3D 海藻酸盐系统中培养了小鼠 CL，该系统可以调整为不同的机械刚度。在目标 1 中，拟议的实验将操纵藻酸盐刚性并确定其对 CL 结构和类固醇生成的影响。具体来说，实验将通过在更严格的藻酸盐条件下培养年轻动物的腺体来复制较老的 CL。相反，救援实验将使用在不太严格的藻酸盐环境中培养的老年动物的 CL 进行。此外，还将通过竞争性抑制整合素与游离RGD肽的结合来探索ECM粘附性和机械力的关系。在该提案的第二个目标中，实验将测试黄体细胞机械转导的可能机制。在许多细胞类型中，Rho/ROCK 信号传导在感知物理环境中发挥着主要作用，并且是 ECM-整合素结合的已知效应器。此外，Rho/ROCK 信号传导可能是 ECM 刚性、整合素结合和黄体细胞结构功能之间的关键联系。实验将评估年轻和年长动物群体中的 Rho/ROCK 信号传导。此外，我们将使用我们的 3D 水凝胶系统和几种途径抑制剂测试以各种硬度培养的 CL 中的 Rho/ROCK 信号传导。通过利用卵巢微环境生物工程的能力，我们可以确定年龄、组织刚性和激素产生交叉点的调节机制。在此过程中，我们将测试一种新的女性生殖衰老理论，并确定可能在体内调节的机制，以改善衰老、绝经前女性的黄体功能。 公共健康相关性：随着年龄的增长，卵巢激素的变化对女性健康起着重要作用，增加了心血管疾病、骨质疏松症和不孕症的风险。该项目的目标是了解与年龄相关的卵巢纤维化如何改变激素的产生。确定卵巢纤维化的后果将改善老年绝经前女性激素相关问题的诊断和治疗。