The Impact of COL6A3 Mutation on Inflammatory Susceptibility in Osteoarthritis

COL6A3 突变对骨关节炎炎症易感性的影响

基本信息

批准号：
10387150
负责人：
Zainab Harissa
金额：
$ 4.68万
依托单位：
WASHINGTON UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-04-01 至 2025-03-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10387150
关键词：
3-Dimensional Affect Area Atomic Force Microscopy Biochemical Biological Biomechanics COL6A3 Cartilage Cartilage Matrix Catabolism Cell Culture Techniques Chemicals Chondrocytes Collagen Type VI DNA Sequence Alteration Degenerative polyarthritis Diffuse Disease Disease Progression Disease model Environment Environmental Risk Factor Equilibrium Exposure to Extracellular Matrix Fluorescence Recovery After Photobleaching Gene Expression Genes Genetic Homeostasis Human IL6 gene In Situ In Vitro Inflammation Inflammation Mediators Inflammatory Interleukin-1 Interleukin-1 beta Ion Channel Knock-out Lead Link Measures Mechanics Mediating Mediator of activation protein Metabolic Modeling Modulus Molecular Molecular Biology Mutate Mutation Onset of illness Pain Pathogenesis Pathway interactions Perception Persons Physiological Predisposition Production Property Proteins Research Risk Risk Factors Role Signal Transduction Testing Therapeutic Uses Tissues Training United States Weight-Bearing state Work antagonist arthropathies cartilage degradation cell type cytokine disability disease-in-a-dish disorder prevention disorder risk early onset environmental stressor exome sequencing exposed human population human model induced pluripotent stem cell inflammatory marker joint inflammation mechanical drive mechanical load mechanical properties mechanical signal mechanical stimulus mechanotransduction novel response symptom management targeted treatment therapeutic development

项目摘要

PROJECT SUMMARY Osteoarthritis (OA) is a painful and debilitating joint disease affecting over 30 million people in the United States alone. OA is characterized by the progressive degeneration of cartilage matrix primarily due to an imbalance of chondrocyte catabolic and anabolic activities arising from genetic, chemical, and mechanical risk factors. Current non-surgical treatments for OA rely simply on managing symptoms primarily owing to a lack of a full understanding of the molecular mechanisms leading to disease onset and pathogenesis. In recent exome sequencing studies, genetic mutation in collagen type VI alpha three (COL6A3) has been identified to increase risk of developing early-onset OA. COL6A3 encodes for one of the proteins that forms collagen type VI (COL6), a critical component of the chondrocyte’s pericellular matrix (PCM). COL6 is a key mediator of the chondrocyte’s transduction of chemical and mechanical cues by maintaining the structural integrity of the PCM. It is likely that mutation in COL6A3 may exacerbate OA by altering PCM properties; however, the mechanisms leading to disease remain unknown. In this proposal, we will determine the molecular link between COL6A3 mutation and OA etiopathogenesis. We hypothesize that COL6A3 mutation increases the effect of inflammation on chondrocyte catabolism and PCM degradation, leading to alterations in the normal chondrocyte metabolic response to mechanical loading thereby initiating a cycle of cartilage degeneration. To complete the proposed studies, we have developed a novel in vitro “OA-in-a-dish” disease model using human induced pluripotent stem cell (hiPSC)-derived chondrocytes gene-edited to introduce the COL6A3 mutation. In specific aim 1, we will assess the effects of proinflammatory cytokines on the biological and mechanical properties of hiPSC-derived cartilage with and without the COL6A3 mutation. Then, in specific aim 2, we will determine the effect of this mutation on chondrocyte mechanotransduction following proinflammatory cytokine treatment. This proposal aims to determine a mechanistic cause for increased predisposition to early on-set OA due to mutation in COL6A3, that can be used for the identification of targets for therapeutic development. Completion of these studies will better define the mechanisms that mechanical, genetic, and biochemical factors act in concert and lead to OA.

项目概要骨关节炎 (OA) 是一种痛苦且令人衰弱的关节疾病，在美国影响着超过 3000 万人单独的 OA 的特点是软骨基质进行性退化，主要是由于软骨基质的不平衡。由遗传、化学和机械风险因素引起的软骨细胞分解代谢和合成代谢活动。 OA 的非手术治疗仅依赖于控制症状，主要依赖于缺乏充分的治疗在最近的外显子组中了解导致疾病发作和发病机制的分子机制。测序研究表明，VI 型胶原蛋白 α 3 (COL6A3) 的基因突变已被确定会增加 COL6A3 编码形成 VI 型胶原蛋白 (COL6) 的一种蛋白质，软骨细胞周基质 (PCM) 的重要组成部分是软骨细胞的关键介质。通过保持 PCM 的结构完整性来传导化学和机械信号。 COL6A3 突变可能通过改变 PCM 特性而加剧 OA；然而，导致 OA 的机制如下：在此提议中，我们将确定 COL6A3 突变与疾病之间的分子联系。我们勇敢地认为 COL6A3 突变会增加炎症对 OA 的影响。软骨细胞分解代谢和 PCM 降解，导致正常软骨细胞发生变化对机械负荷的代谢反应，从而启动软骨退化的循环。完成拟议的研究，我们利用人类开发了一种新型体外“培养皿中的 OA”疾病模型对诱导多能干细胞 (hiPSC) 衍生的软骨细胞进行基因编辑，引入 COL6A3 突变。具体目标1，我们将评估促炎细胞因子对生物和力学的影响具有和不具有 COL6A3 突变的 hiPSC 衍生软骨的特性然后，在具体目标 2 中，我们将进行研究。确定该突变对促炎细胞因子后软骨细胞机械转导的影响该治疗方案旨在确定早发 OA 易感性增加的机制原因。由于 COL6A3 突变，可用于鉴定治疗开发的靶点。完成这些研究将更好地确定机械、遗传和生化因素的机制一致行动并导致OA。