Menopausal Knee-ds: Elucidating mechanisms and treatments for knee osteoarthritis

项目摘要/摘要 背景：截至2020年，估计有6.541亿成年人患有膝骨性关节炎(KOA)，女性 绝经后女性患膝关节骨性关节炎的可能性几乎是男性的两倍。尽管如此，大多数动物 关于膝关节骨性关节炎的研究只包括男性，在少数利用女性的研究中，更年期通常不包括在内， 因为在非灵长类动物中这不是一个自然发生的过程。因此，针对这一问题的文献很少 了解与绝经相关的膝骨性关节炎的机制和相应的治疗缺失 专门针对绝经后膝关节骨性关节炎患者的干预措施。到目前为止，作为我博士工作的一部分，我 通过给中年雌性小鼠注射4-氨基丁酸建立了一种化学诱导的更年期模型。 乙烯基环己烯二环氧化物(VCD)。我已经确认这个模型显示了更年期的表型，包括 这些小鼠比年龄匹配的非更年期小鼠表现出更严重的膝骨性关节炎。 本研究会的研究目标是探讨绝经后膝关节骨性关节炎的发病机制。 我们的VCD模型(特定目标1)，并采用合成生物学技术为新的 绝经引起的膝骨性关节炎的治疗方式和(具体目标2)。 具体目标1：E3泛素连接酶的一个组分最近在GWAS荟萃分析中被确定为唯一的 女性骨性关节炎的贡献者。因此，我将询问雌激素调节的泛素蛋白水解酶在 使用我们的VCD绝经模型调节绝经期诱发的KOA。泛素蛋白分解的变化 围绝经期、绝经期和雌激素治疗的信号将在体内和体内被量化。 体外培养。我们假设(1)绝经诱导会扰乱泛素蛋白分解活性和(2) 更年期早期开始的雌激素治疗将恢复泛素蛋白分解信号，并最终 绝经期的膝骨性关节炎。 具体目标2：我将设计一个雌激素调节的控制电路，在体外调节感兴趣基因(GOI)。 雌激素的显著下降是与更年期有关的主要生理变化，而雌激素- 被抑制的启动子将被设计成开启电路。GOI候选人将从以前的 研究并将进行系统测试，以确定调节软骨生成的理想性。我们假设 雌激素调节的遗传控制电路将在体外减弱软骨细胞的健康。 影响：为了支持我的奖学金，我组建了一个多学科团队，拥有KOA、更年期、 衰老和合成生物学。我已经和我的指导团队制定了战略，以设计一项严格的培训计划， 将利用我的赞助商的新机构提供的非凡的研究环境 (哈佛医学院斯波尔丁康复学院)和匹兹堡大学，在那里我仍然是一名全职 学生。这笔奖学金将推动我朝着成为一名实践的内科科学家的长期目标前进 在女性关节疾病和损伤方面的整形外科和研究领先者。

PROJECT SUMMARY/ABSTRACT Background: As of 2020, an estimated 654.1 million adults live with knee osteoarthritis (KOA), and women who are post-menopausal are nearly twice as likely to develop KOA compared to men. Despite this, most animal studies on KOA include only males, and, of the few studies utilizing females, menopause is typically not included, as it is not a naturally occurring process in non-primates. As such, there is a paucity of literature aimed at understanding mechanisms of menopause associated KOA and a corresponding absence of treatment interventions specifically for post-menopausal people with KOA. As a part of my PhD work thus far, I have developed a chemically-induced menopause model by injecting middle-aged female mice with 4- vinylcyclohexene diepoxide (VCD). I have confirmed this model displays a menopausal phenotype, including perimenopause, and that these mice display more severe KOA than age-matched, non-menopausal mice. The research goals of this fellowship are to interrogate mechanisms of menopause-induced KOA using our VCD model (Specific Aim 1) and employ synthetic biology techniques to lay the groundwork for new treatment modalities for menopause-indued KOA and (Specific Aim 2). Specific Aim 1: A component of E3 ubiquitin ligase was recently identified in a GWAS meta-analysis as a unique contributor to OA in women. As such, I will interrogate the role of estradiol-regulated, ubiquitin proteolysis in mediating menopause-induced KOA using our VCD menopause model. Changes in ubiquitin proteolysis signaling across perimenopause, menopause, and with estradiol treatment will be quantified both in vivo and in vitro. We hypothesize that (1) menopause induction will disrupt ubiquitin proteolysis activity and (2) estradiol treatment started early in menopause will restore ubiquitin proteolysis signaling and ultimately quinch menopausal KOA. Specific Aim 2: I will design an estradiol-regulated controls circuit to modulate a gene of interest (GOI) in vitro. A significant drop in estradiol is a principal physiological change associated with menopause, and an estradiol- repressed promoter will be designed to turn the circuit on. GOI candidates will be generated from previous studies and will be systematically tested to determine ideality for mediating chondrogenicity. We hypothesize that a genetic controls circuit modulated by estradiol will attenuate chondrocyte health in vitro. Impact: To support my fellowship, I have assembled a multidisciplinary team with expertise in KOA, menopause, aging, and synthetic biology. I have strategized with my mentoring team to design a rigorous training plan that will take advantage of the extraordinary research environments offered at my sponsor’s new institution (Spaulding Rehabilitation, Harvard Medical School) and the University of Pittsburgh, where I remain a full-time student. This fellowship will propel me towards my long-term goals of being a physician-scientist who practices orthopedic surgery and research leader in joint diseases and injuries presenting in women.