Identification of Novel Markers for Systemic Sclerosis Employing Proteomics

利用蛋白质组学鉴定系统性硬化症的新标记物

• 批准号: 8125088
• 负责人: SERGIO A JIMENEZ
• 金额: $ 20.09万
• 依托单位: THOMAS JEFFERSON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-08-09 至 2012-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8125088
• 关键词: Affect; Age; Antibodies; Biological Markers; Biometry; Biopsy; Clinical; Clinical Management; Clinical Trials; Collagen; Connective Tissue; Coupled; Custom; Cutaneous; Data; Deposition; Dermal; Development; Diagnosis; Diffuse; Disease; Early Diagnosis; Employee Strikes; Enzyme-Linked Immunosorbent Assay; Fibroblasts; Fibrosis; Fluorescence; Gender; Immunoassay; Indirect Immunofluorescence; Individual; Lead; Mass Spectrum Analysis; Measures; Methods; Organ; Outcome; Outcome Measure; Patients; Phenotype; Process; Progressive Disease; Protein Analysis; Proteins; Proteomics; Questionnaires; Race; Research Personnel; Rheumatoid Arthritis; SF-36; Sampling; Serum; Set protein; Severities; Signal Transduction; Skin; Specificity; Systemic Lupus Erythematosus; Systemic Scleroderma; Testing; Therapeutic Agents; Tissues; Validation; Western Blotting; base; design; macromolecule; multiple reaction monitoring; novel; novel marker; outcome forecast; prognostic; public health relevance; response; therapeutic evaluation

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.

描述（由申请人提供）：系统性硬化症（Systemic Sclerosis, 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最明显和最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。可用于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无法用于SSc。开发反映组织纤维化程度和严重程度的客观可靠的标志物，将为确定临床试验中特定治疗的疗效提供宝贵的帮助，提供客观的方法，提供无偏见的信息，从而减少获得统计显著性所需的患者数量。在SSc患者的临床管理中，这些标志物的可用性也将对评估疾病调节剂的治疗反应具有重要价值。目的/假设：我们的初步数据有力地支持了这样一个前提，即通过对SSc成纤维细胞分泌组的蛋白质组学分析可以识别出反映成纤维细胞活化及其纤维化生物合成表型的程度和严重程度的蛋白质，并且这些蛋白质在SSc患者的皮肤活检和血清中可以检测到。基于此前提，我们建议在本申请中检验假设：“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中表达差异最大的蛋白质，然后在SSc患者的血清中验证结果，将导致鉴定高度可靠和特异性的生物标志物，可用于早期诊断和评估SSc组织纤维化的程度和严重程度”。我们提出以下具体目标来验证这一假设：具体目标1：通过蛋白质组学分析，与正常真皮成纤维细胞的分泌组相比，鉴定出SSc成纤维细胞分泌组中表达差异最大的蛋白质。目的2：通过ELISA或多反应监测（MRM）验证蛋白质组学分析鉴定的蛋白作为SSc患者血清中SSc生物标志物的特异性。特异性目的3：通过建立与相关临床参数的相关性来确定鉴定蛋白的临床价值。该项目的成功结果将导致确定SSc的新结果措施，这将对SSc临床试验的设计和分析以及受这种无法治愈、致残且往往致命的疾病影响的患者的临床管理产生变革性影响。

项目成果

Altered MCM protein levels and autophagic flux in aged and systemic sclerosis dermal fibroblasts.

• DOI: 10.1038/jid.2014.69
• 发表时间: 2014-09
• 期刊: JOURNAL OF INVESTIGATIVE DERMATOLOGY
• 影响因子: 6.5
• 作者: Dumit, Veronica I.;Kuettner, Victoria;Kaeppler, Jakob;Piera-Velazquez, Sonsoles;Jimenez, Sergio A.;Bruckner-Tuderman, Leena;Uitto, Jouni;Dengjel, Joern
• 通讯作者: Dengjel, Joern