Defining Common Molecular Parameters For Onset and Progression of Osteoarthritis

描述(由申请人提供)：这项RC4在主题领域1--应用基因组学和其他高通量技术--的申请是一项综合的、多学科的战略，旨在实现在确定支撑骨关节炎(OA)的常见生物分子机制方面的重大“飞跃”。针对特定靶点的合理治疗策略严重缺乏，这些靶点决定了OA疾病的发病和严重程度。传统的先前努力主要集中在单个调节分子上，从而阻碍了我们破解显然是多因素疾病或疾病组的发展背后的谜团的能力。我们的研究团队之前的成就导致了新的小鼠OA疾病模型的开发，同样重要的是，还发现了特定的转录因子和上游信号分子在软骨细胞应激和分化中的作用。其中包括核因子-B、Runx2、ESE-1/Elf3、GADD452和DDR2，它们中的每一个都会影响软骨细胞中主要的胶原降解酶--基质金属蛋白酶-13的表达。因此，我们的发现提供了一个概念性的框架，将被用来分析多个小鼠OA疾病模型中基于基因组和磷酸蛋白质组的无偏见筛选。我们假设，应激或炎症诱导的信号不仅有助于骨关节炎不可逆转的关节损伤(进展)，而且重要的是，在启动/发病阶段，关节软骨中的软骨细胞离开其自然生长和分化停滞状态。我们将使用三种精心挑选的小鼠OA模型，每一种模型都通过软骨细胞功能和生理上的不同固有变化影响OA疾病的发展：(1)推动软骨细胞向肥大分化的关键转录因子Runx2的表达减少；(2)典型的NF-：B信号的丢失，这是协调对细胞内和细胞外应激做出不同反应的主要途径；以及(3)CHO/突变小鼠的XI型胶原单倍性不足。这3个小鼠OA模型中的每一个都将首次综合使用microRNA(MiRNA)和基因表达阵列、磷酸蛋白蛋白质组学(最初专注于NF-：B、MAPK/ERK、p38、JNK、JAK/STAT和Tak1/Smad级联)以及系统水平的生物信息学，以确定在遗传和创伤后小鼠OA中最常激活的关键网络和靶点。重要的是，后一种新的数据将被用来询问现有的来自人类骨性关节炎软骨、滑膜和滑液的基因组或蛋白质组数据集，以揭示在骨性关节炎疾病发生和发展过程中保守的、临床上相关的靶点。 公共卫生相关性：这份RC4提案(主题领域1)涉及一种多学科方法，使用强大的基因表达、miRNA和单细胞磷酸蛋白质组筛选，将给出影响骨性关节炎疾病发生和发展的软骨重要调控网络的全面和综合图景。四个不同机构的多个PI的创新和协调努力将在3个创伤后或遗传性骨关节炎小鼠模型中发现新的“共同效应器”和关键网络。此外，对现有的人类骨性关节炎数据集的询问将在“整个关节”的背景下产生临床相关的影响，并发现新的治疗靶点。

DESCRIPTION (provided by applicant): This RC4 application in thematic area 1-- Applying Genomics and Other High Throughput Technologies-is an integrated, multi-disciplinary strategy to achieve a major "leap forward" in defining common biomolecular mechanisms underpinning osteoarthritis (OA). Rational therapeutic strategies against specific targets that determine the onset and severity of OA disease are severely lacking. Traditional prior efforts have primarily focused on single regulatory molecules, thereby hindering our ability to decipher the puzzle underlying the development of what is clearly a multifactorial disease or group of diseases. Prior achievements of our team of investigators have led to the development of novel murine models of OA disease and, of equal importance, also uncovered roles of specific transcription factors and upstream signaling molecules contributing to chondrocyte stress and differentiation. Among these are NF-B, Runx2, ESE-1/Elf3, GADD452, and DDR2, each of which impact on the expression of MMP-13, the major collagen-degrading enzyme in chondrocytes. Thus, our findings provide a conceptual framework that will be exploited to analyze unbiased genome-based and phospho-proteome screens across multiple mouse OA disease models. We hypothesize that stress- or inflammation-induced signals not only contribute to IRREVERSIBLE joint damage (progression) in OA, but importantly, also to the initiation/onset phase, wherein chondrocytes in articular cartilage leave their natural growth- and differentiation-arrested state. We will employ 3 carefully chosen murine OA models, each of which affect OA disease development by different inherent changes in chondrocyte function and physiology: (1) diminished expression of Runx2, a pivotal transcription factor that drives chondrocyte differentiation towards hypertrophy, and (2) loss of canonical NF-:B signaling, the major pathway orchestrating diverse responses to intracellular and extracellular stress, and (3) haploinsufficiency of type XI collagen in the cho/+ mutant mouse. Each of these 3 murine OA models will be subjected to the first integrated use of vanguard technologies of microRNA (miRNA) and gene expression arrays, phospho-protein proteomics (focusing initially on NF-:B, MAPK/ERK, p38, JNK, JAK/STAT, and Tak1/Smad cascades), and systems-level bioinformatics to define key networks and targets most commonly activated in genetic and post-traumatic murine OA. Importantly, the latter novel data will then be used to interrogate existing genome or proteome datasets from human OA cartilage, synovia and synovial fluids to uncover the conserved, clinically relevant targets in the context of the "whole joint" in OA disease development and progression. PUBLIC HEALTH RELEVANCE: This RC4 proposal (Thematic Area 1) involves a multi-disciplinary approach using powerful gene expression, miRNA, and single cell phospho-proteome screens that will give a comprehensive and integrated picture of the important regulatory networks in cartilage that impact on OA disease onset and progression. The innovative and coordinated efforts of multiple PIs at four different institutions will uncover novel 'common effectors' and critical networks across 3 murine models with post-traumatic or genetic OA. Moreover, interrogation against available human OA datasets will have clinically relevant impact in the context of the "whole joint" and uncover new therapeutic targets.