Defining Common Molecular Parameters For Onset and Progression of Osteoarthritis

描述（由申请人提供）：主题领域1的RC4申请-应用基因组学和其他高通量技术-是一项综合的多学科战略，旨在实现定义骨关节炎（OA）基础的常见生物分子机制的重大“飞跃”。目前严重缺乏针对确定OA发病和严重程度的特定靶点的合理治疗策略。传统的先前的努力主要集中在单一的调节分子上，从而阻碍了我们破译明显是一种多因素疾病或一组疾病发展背后的谜团的能力。我们的研究团队先前的成就导致了OA疾病的新型小鼠模型的发展，同样重要的是，也揭示了特定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NF-B、Runx2、ESE-1/Elf3、GADD452和DDR2影响软骨细胞中主要的胶原降解酶MMP-13的表达。因此，我们的研究结果提供了一个概念性框架，将用于分析多种小鼠OA疾病模型的无偏倚基因组和磷酸化蛋白质组筛选。我们假设，应激或炎症诱导的信号不仅有助于OA中不可逆的关节损伤（进展），而且重要的是，也有助于起始/发病阶段，其中关节软骨中的软骨细胞离开其自然生长和分化停滞状态。我们将采用3种精心挑选的小鼠OA模型，每种模型都通过不同的软骨细胞功能和生理固有变化影响OA疾病的发展：(1)驱动软骨细胞向肥大分化的关键转录因子Runx2的表达减少，(2)规范的NF-:B信号通路的缺失，这是协调细胞内和细胞外应激的多种反应的主要途径，(3)cho/+突变小鼠XI型胶原的单倍不足。这3种小鼠OA模型中的每一种都将首次综合使用microRNA （miRNA）和基因表达阵列、磷酸化蛋白蛋白质组学（最初专注于NF-:B、MAPK/ERK、p38、JNK、JAK/STAT和Tak1/Smad级联）和系统级生物信息学等先进技术，以确定遗传和创伤后小鼠OA中最常见的关键网络和靶点。重要的是，后者的新数据将用于询问现有的来自人类OA软骨、滑膜和滑膜液的基因组或蛋白质组数据集，以揭示OA疾病发展和进展中“全关节”背景下保守的、临床相关的靶点。

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.