Modeling ovarian aging phenotype in mechanically tuned 3D matrices

在机械调谐 3D 矩阵中模拟卵巢衰老表型

• 批准号: 10213599
• 负责人: Emma Gargus
• 金额: $ 3.52万
• 依托单位: NORTHWESTERN UNIVERSITY AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-09-01 至 2022-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10213599
• 关键词: 3-Dimensional; 3D Print; Address; Adhesions; Age; Aging; Animals; Behavior; Biological; Biological Assay; Biology; Biomedical Engineering; Cardiovascular Diseases; Cell model; Cells; Coupled; Critical Thinking; Cues; Cytoskeleton; Data; Diagnosis; Disease; Engineering; Ensure; Environment; Estrogens; Extracellular Matrix; Family; Female; Female infertility; Fibrosis; Functional disorder; Gelatin; Goals; Growth; Health; Hormone secretion; Hormones; Hyperplasia; In Vitro; Infertility; Ink; Integrins; Link; Measures; Mechanics; Mediating; Menopause; Mitogen-Activated Protein Kinases; Modeling; Molecular; Mus; Oligomenorrhea; Oocytes; Oranges; Organ; Osteoporosis; Ovarian; Ovarian Carcinoma; Ovarian Follicle; Ovarian aging; Ovarian hormone; Ovary; Pathway interactions; Phenocopy; Phenotype; Physiological; Play; Polycystic Ovary Syndrome; Premenopause; Process; Production; Progesterone; Reproducibility; Research; Role; Signal Pathway; Signal Transduction; Steroid biosynthesis; Stromal Cells; Structure; Techniques; Testing; Tissues; Woman; Women' s Health; age related; body system; cardiovascular disorder risk; cell type; cohort; density; drug development; drug discovery; egg; experimental study; female fertility; gonad function; granulosa cell; improved; in vitro Model; in vivo; inhibitor/antagonist; mechanical properties; mechanotransduction; mouse model; novel; older women; oocyte quality; ovarian dysfunction; reproductive; response; rho; rho GTP-Binding Proteins; scaffold; skills

PROJECT SUMMARY Ovarian aging is associated with fibrosis, a changing female hormone profile, and a decline in oocyte quality and quantity, resulting in sequelae such as cardiovascular disease, osteoporosis, and infertility. Even before menopause, there are significant age-related changes in hormone production and oocyte quality. Ovarian aging is a fibrotic process involving dramatic extracellular matrix remodeling, resulting in an increasingly rigid microenvironment. While it is known that ECM-derived signals regulate steroidogenesis, the causative relationship between matrix mechanics and function has not previously been demonstrated. Thus, in the studies proposed herein, I will test the hypothesis that mechanical changes in the follicle microenvironment cause a progressive decline in hormone production and egg quality in an aging mouse model. In preliminary studies, we have demonstrated that 3D-printed gelatin scaffolds – which can be experimentally tuned to different rigidities— support ovarian follicle survival, growth, and function. In Aim 1, I will experimentally manipulate the stiffness of the 3D printed gelatin scaffolds in order to define the effects of rigidity on follicle structure and function and oocyte quality. Specifically, I hypothesize that I can phenocopy older follicles (i.e. diminished hormone production and oocyte quality) by culturing follicles isolated from younger mice in more rigid scaffolds. Conversely, I will perform a rescue experiment with follicles from older animals cultured in a softer scaffold. In Aim 2, proposed experiments will test a possible mechanism of follicle mechanotransduction. In many cell types, Rho/ROCK signaling is responsible for converting mechanical cues into biological response. Moreover, in preliminary studies, we demonstrate that Rho, phospho-Rho, and ROCK are present in discrete follicles in the murine ovary, indicating that Rho signaling is an available molecular mechanism for follicle mechanotransduction. I will assay Rho/ROCK signaling in reproductively younger and older mouse cohorts. Additionally, I will test Rho/ROCK signaling in follicles cultured in 3D printed scaffolds of various rigidities and in the presence of pathway inhibitors. These studies will test my overarching hypothesis that the mechanical properties of the ovarian matrix play a role in age-related ovarian dysfunction (decline in hormone production and egg quality), possibly via mechanosensitive Rho/ROCK signaling. Moreover, these mechanically-tunable 3D-printed scaffolds represent novel in vitro models of ovarian aging and provide a platform for drug discovery and development that may revolutionize the treatment of age-related female infertility.

项目概要 卵巢衰老与纤维化、女性激素变化和卵母细胞质量下降有关 和数量，导致心血管疾病、骨质疏松、不孕不育等后遗症。甚至之前 更年期，激素产生和卵母细胞质量会出现与年龄相关的显着变化。卵巢 衰老是一个纤维化过程，涉及细胞外基质的剧烈重塑，导致细胞外基质变得越来越僵硬。 微环境。虽然众所周知 ECM 衍生信号可调节类固醇生成，但其致病原因 矩阵力学和函数之间的关系以前尚未得到证明。因此，在 在本文提出的研究中，我将检验卵泡微环境的机械变化的假设 导致衰老小鼠模型中激素产生和卵子质量逐渐下降。在初步 研究中，我们证明了 3D 打印的明胶支架——可以通过实验调整 不同的硬度——支持卵泡的存活、生长和功能。在目标 1 中，我将通过实验 操纵 3D 打印明胶支架的硬度，以确定硬度对卵泡的影响 结构和功能以及卵母细胞质量。具体来说，我假设我可以对较旧的卵泡进行表型复制（即 通过在更多的环境中培养从年轻小鼠中分离的卵泡来减少激素产生和卵母细胞质量） 刚性脚手架。相反，我将用在一个实验室中培养的老年动物的卵泡进行救援实验。 更软的脚手架。在目标 2 中，拟议的实验将测试卵泡的可能机制 力传导。在许多细胞类型中，Rho/ROCK 信号传导负责转换机械信号 进入生物反应。此外，在初步研究中，我们证明 Rho、磷酸-Rho 和 ROCK 存在于小鼠卵巢的离散卵泡中，表明 Rho 信号传导是一种可用的分子 卵泡机械传导机制。我将在更年轻和更年轻的生殖细胞中检测 Rho/ROCK 信号传导 年长的小鼠群体。此外，我将测试在 3D 打印支架中培养的卵泡中的 Rho/ROCK 信号传导 各种刚性和存在途径抑制剂的情况下。这些研究将检验我的总体假设 卵巢基质的机械特性在与年龄相关的卵巢功能障碍（卵巢功能下降）中发挥作用 激素产生和卵子质量），可能通过机械敏感的 Rho/ROCK 信号传导。而且，这些 机械可调的 3D 打印支架代表了卵巢衰老的新型体外模型，并提供了 药物发现和开发平台可能会彻底改变与年龄相关的女性的治疗 不孕症。