Identification of Novel Markers for Systemic Sclerosis Employing Proteomics

描述（申请人提供）：系统性硬化症（SSc）是一种病因不明的疾病，其特征是皮肤和多个内脏器官中胶原蛋白和其他结缔组织大分子过度沉积。SSc最明显和几乎普遍的临床特征与皮肤、微血管和许多内脏器官的进行性纤维化有关。对于SSc，尚不能获得允许早期诊断和评估疾病活动性或具有预测预后价值的经过充分验证的生物标志物。客观和可靠的指标反映组织纤维化的程度和严重程度的发展将是非常宝贵的援助，在确定一个给定的治疗在临床试验中的疗效，通过提供一个客观的方法，提供公正的信息，允许在获得统计学意义所需的患者人数减少。这些标志物的可用性也将在评估SSc患者临床管理中对疾病修饰剂的治疗反应方面具有重要价值。 目的/假设：我们的初步数据强烈支持的前提，即蛋白质组学分析的SSc成纤维细胞分泌组可以识别蛋白质，反映成纤维细胞活化和程度和严重程度的促纤维化生物合成表型，这些蛋白质是可检测的皮肤活检和血清的SSc患者。基于这一前提，我们提出在本申请中测试以下假设："通过蛋白质组学研究鉴定SSc成纤维细胞分泌组中与正常成纤维细胞分泌组相比最差异表达的蛋白质，然后验证SSc患者血清中的结果，将导致鉴定高度可靠和特异的生物标志物，其可用于早期诊断和评估SSc中组织纤维化的程度和严重程度”。 我们建议用以下具体目的来检验这一假设：具体目的1：通过蛋白质组学分析鉴定SSc成纤维细胞分泌组中与正常真皮成纤维细胞分泌组相比差异表达最高的蛋白质。具体目标2：通过ELISA或多反应监测（MRM）验证通过蛋白质组学分析鉴定的蛋白质作为SSc患者血清中SSc生物标志物的特异性。具体目的3：通过建立与相关临床参数的相关性来确定所鉴定蛋白质的临床价值。 该项目的成功结果将导致确定SSc的新结局指标，这将对SSc临床试验的设计和分析以及受这种无法治愈，致残且通常致命的疾病影响的患者的临床管理产生转变性影响。 公共卫生相关性：采用蛋白质组学鉴定系统性硬化症的新型生物标志物将导致系统性硬化症的新型结局指标的鉴定，这将对系统性硬化症临床试验的设计和分析以及受这种不可治愈、致残且通常致命的疾病影响的患者的临床管理产生转化性影响。

DESCRIPTION (provided by applicant): Systemic Sclerosis (SSc) is a disease of unknown origin characterized by excessive deposition of collagen and other connective tissue macromolecules in skin and multiple internal organs. The most apparent and almost universal clinical features of SSc are related to the progressive fibrosis of the skin, the microvasculature, and numerous internal organs. Well validated biomarkers that allow early diagnosis and assessment of disease activity or that carry a predictive prognostic value are not available for SSc. The development of objective and reliable markers reflecting the extent and severity of tissue fibrosis would be of invaluable assistance in determining the efficacy of a given treatment in clinical trials by offering an objective method that provides unbiased information which allows a reduction in the number of patients required to obtain statistical significance. Availability of such markers will also be of substantial value in the evaluation of therapeutic responses to disease modifying agents in the clinical management of SSc patients. Objective/Hypothesis: Our preliminary data strongly support the premise that proteomic analysis of the SSc fibroblast secretome can identify proteins that reflect fibroblast activation and the extent and severity of their profibrotic biosynthetic phenotype, and that these proteins are detectable in skin biopsies and sera of SSc patients. Based on this premise, we propose to test in this application the hypothesis that: "the identification by proteomic studies of the most differentially expressed proteins in the SSc fibroblast secretome compared to the secretome of normal fibroblasts followed by the validation of the results in serum from SSc patients will lead to the identification of highly reliable and specific biomarkers that can be used for early diagnosis and for assessment of extent and severity of tissue fibrosis in SSc". We propose to test the hypothesis with the following Specific Aims: SPECIFIC AIM 1: To identify by proteomic analysis the most highly differentially expressed proteins in the SSc fibroblast secretome compared to the secretome of normal dermal fibroblasts. SPECIFIC AIM 2: To validate the specificity of the proteins identified by proteomic analysis as a biomarker for SSc in sera from SSc patients by ELISA or Multiple Reaction Monitoring (MRM). SPECIFIC AIM 3: To determine the clinical value of the identified proteins by establishing correlations with relevant clinical parameters. The successful outcome of this project will lead to the identification of novel outcome measures for SSc that will have a transforming impact in the design and analysis of clinical trials for SSc and in the clinical management of patients affected by this incurable, disabling, and often fatal disease. PUBLIC HEALTH RELEVANCE: The identification of novel biomarkers for Systemic Sclerosis employing proteomics will lead to the identification of novel outcome measures for Systemic Sclerosis that will have a transforming impact in the design and analysis of clinical trials for Systemic Sclerosis and in the clinical management of patients affected by this incurable, disabling, and often fatal disease.