CTGF-interacting proteins in scleroderma lung fibrosis

硬皮病肺纤维化中的 CTGF 相互作用蛋白

• 批准号: 7616777
• 负责人: GALINA S BOGATKEVICH
• 金额: $ 12.54万
• 依托单位: MEDICAL UNIVERSITY OF SOUTH CAROLINA
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-05-01 至 2010-10-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7616777
• 关键词: Amino Acid Sequence; Area; Award; Binding; Binding Proteins; Biochemical; Biology; Cause of Death; Cell Surface Proteins; Cessation of life; Complex; Data; Detection; Development; Disease; Doctor of Philosophy; Employee Strikes; Evaluation; Fibroblasts; Fibrosis; Goals; Laboratories; Lung; Lung diseases; Mass Spectrum Analysis; Pathogenesis; Patients; Peptide Sequence Determination; Pharmacology; Play; Process; Progressive Disease; Proteins; Proteomics; Pulmonary Fibrosis; Receptor Signaling; Research; Research Project Grants; Rheumatology; Role; Scleroderma; Signal Transduction; Spottings; Techniques; Technology; Testing; Tissues; Training; Two-Dimensional Gel Electrophoresis; base; career; connective tissue growth factor; crosslink; cytokine; design; effective therapy; experience; extracellular; gel electrophoresis; instrument; instrumentation; interest; interstitial; molecular marker; new therapeutic target; novel; protein complex; protein crosslink; protein protein interaction; receptor; reconstruction; sensor; skills

DESCRIPTION (provided by applicant): This proposal provides a means of laying the groundwork for the candidate's career in scleroderma research. Her formal Ph.D. training was primarily in pharmacology and receptor signaling. She has developed an interest in proteomics technologies and wants to apply them to scleroderma research. In the course of this award period, the candidate expects to develop skills in the areas of rheumatology, biology of scleroderma fibroblasts, mass spectrometry, and proteomics that are much needed, through both didactic and laboratory experience. Then, using these new skills in combination with her receptor pharmacology and signal transduction background, she will develop independent research projects investigating the fundamental mechanisms of scleroderma fibrosis. Toward these ends, a proposal has been prepared which should provide ample opportunity to develop new skills and to obtain the theoretical background in scleroderma research required for the candidate's long-term goals. The research plan of the proposal is based on the observations that connective tissue growth factor (CTGF) Specifically binds numerous proteins in lung fibroblasts derived from scleroderma patients. CTGF is highly expressed in many fibrotic disorders and is considered to be a molecular marker of fibrosis. Interstitial lung fibrosis strikes up to 80% of scleroderma patients and is now the leading cause of death. Since scleroderma lung fibroblasts not only synthesize increased amounts of CTGF, but also contain proteins that specifically interact with CTGF, we postulate that such interactions are critical for the pathogenesis and expansion of scleroderma pulmonary fibrosis. Our research plan aims to identify the CTGF-interacting proteins and to define the components of a signal transduction complex for CTGF in scleroderma lung fibroblasts. This project will involve a combination of biochemical and proteomic techniques applied to the detailed reconstruction of CTGF's microenvironment in scleroderma lung fibroblasts. Part of the proposed studies will be carried out with BIAcore 3000, which allows detection of even transient binding of proteins. The use of this commercial instrument in this manner is a novel technique, which is not commonly available. An additional factor that makes this proposal particularly unique is the availability of the well-equipped Mass Spectrometry Facility, which offers instrumentation to sequence proteins at picomolar levels and lower. Cutting-edge proteomics technologies will allow us to identify CTGF-interacting proteins resolved via ID or 2D gel electrophoresis or eluted from the BIAcore sensor chip, as well as to spot the complex of protein cross linked to CTGF. The identified protein(s) might serve then as potential target(s) in the treatment of scleroderma pulmonary fibrosis for which no proven, effective therapy currently exists.

描述（由申请人提供）：该提案提供了一种为候选人在硬皮病研究中的职业生涯奠定基础的方法。她的正式博士学位。培训主要是药理学和受体信号传导。她对蛋白质组学技术产生了兴趣，并希望将其应用于硬皮病研究。在此奖项期间，候选人希望通过教学和实验室经验，培养急需的流变学，硬皮病成纤维细胞生物学，质谱和蛋白质组学领域的技能。然后，利用这些新技能结合她的受体药理学和信号转导背景，她将开发独立的研究项目，调查硬皮病纤维化的基本机制。为此，已经准备了一份提案，该提案应提供充分的机会来发展新技能，并获得候选人长期目标所需的硬皮病研究理论背景。 该提案的研究计划是基于结缔组织生长因子（CTGF）特异性结合来自硬皮病患者的肺成纤维细胞中的许多蛋白质的观察。CTGF在许多纤维化疾病中高度表达，并且被认为是纤维化的分子标志物。间质性肺纤维化侵袭高达80%的硬皮病患者，并且现在是死亡的主要原因。由于硬皮病肺成纤维细胞不仅合成增加量的CTGF，而且还含有与CTGF特异性相互作用的蛋白质，我们假设这种相互作用对硬皮病肺纤维化的发病机制和扩展至关重要。我们的研究计划旨在确定CTGF相互作用的蛋白质，并确定在硬皮病肺成纤维细胞的CTGF信号转导复合物的组成部分。 该项目将涉及生物化学和蛋白质组学技术相结合，应用于硬皮病肺成纤维细胞中CTGF微环境的详细重建。部分拟议研究将使用BIAcore 3000进行，该仪器可检测蛋白质的瞬时结合。以这种方式使用这种商业仪器是一种新颖的技术，这是不常见的。另一个使这项建议特别独特的因素是设备齐全的质谱设施的可用性，该设施提供了在皮摩尔水平和更低水平上对蛋白质进行测序的仪器。尖端的蛋白质组学技术将使我们能够识别通过ID或2D凝胶电泳解析或从BIAcore传感器芯片洗脱的CTGF相互作用蛋白质，以及发现与CTGF交联的蛋白质复合物。然后，所鉴定的蛋白质可以用作治疗硬皮病肺纤维化的潜在靶标，目前还不存在针对硬皮病肺纤维化的经证实的有效疗法。

项目成果

Antiinflammatory and antifibrotic effects of the oral direct thrombin inhibitor dabigatran etexilate in a murine model of interstitial lung disease.

• DOI: 10.1002/art.30255
• 发表时间: 2011-05
• 期刊: ARTHRITIS AND RHEUMATISM
• 作者: Bogatkevich, Galina S.;Ludwicka-Bradley, Anna;Nietert, Paul J.;Akter, Tanjina;van Ryn, Joanne;Silver, Richard M.
• 通讯作者: Silver, Richard M.

Coagulation and autoimmunity in scleroderma interstitial lung disease.

• DOI: 10.1016/j.semarthrit.2010.10.002
• 发表时间: 2011-10
• 期刊: Seminars in arthritis and rheumatism
• 影响因子: 5
• 作者: Ludwicka-Bradley A;Silver RM;Bogatkevich GS
• 通讯作者: Bogatkevich GS