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: 8308731
• 负责人: Robin M. Skory
• 金额: $ 3.42万
• 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 2013-07-05
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8308731
• 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; young woman

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.

描述（由申请人提供）：卵巢衰老永久改变雌性激素的特征，引起后遗症，例如不育，骨质疏松症和心血管疾病。在更年期之前，在月经周期的两个阶段，激素产生与年龄有关的变化。与年轻的妇女相比，老年妇女的黄体相激素水平较低，这是由黄体（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戏剧性基质重塑的纤维化过程，可提高微环境的刚度。尽管众所周知，ECM衍生的信号调节霉素生成，但以前尚未显示微环境刚度与激素产生之间的致病联系。因此，在本文提出的研究中，我们将检验以下假设：卵巢物理环境的变化会导致衰老小鼠模型的进行性黄体相激素的下降。在初步研究中，我们已经在新型的3D藻酸盐系统中培养了鼠CL，可以调整为不同的机械刚性。在AIM 1中，提出的实验将操纵藻酸盐的刚度，并定义对CL结构和类固醇生成的影响。具体而言，实验将通过在更刚性藻酸盐条件下培养幼小动物的腺体来培养较老的CL。相反，将使用在不太刚性藻酸盐环境中培养的老年动物的CL进行救援实验。另外，通过竞争性抑制与游离RGD肽的整联蛋白结合，将探索ECM粘附性和机械力的关系。在该提案的第二个目的中，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起着主要作用，并且是ECM-整合蛋白参与的已知效应子。此外，RHO/ROCK信号传导可能是ECM刚度，整合素结合和黄体细胞结构功能之间的关键联系。实验将评估年轻动物队列和年龄较大动物队列中的RHO/岩石信号传导。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在各种刚性中培养的CLS中的Rho/Rock信号传导。通过利用生物工程的卵巢微环境的能力，我们可以在年龄，组织刚性和激素产生的交叉点上确定调节机制。这样一来，我们将测试一种新的女性生殖衰老理论，并鉴定可能在体内调节的机制，以改善衰老的绝经前妇女的黄体功能。