Mechanisms of Steroid Resistance in Airway Smooth Muscle Cells

气道平滑肌细胞类固醇抵抗机制

基本信息

批准号：
8010971
负责人：
OMAR TLIBA
金额：
$ 24.8万
依托单位：
THOMAS JEFFERSON UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2007
资助国家：
美国
起止时间：
2007-08-15 至 2012-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8010971
关键词：
Accounting Address Asthma Award Binding Binding Sites Biological Assay Cell Adhesion Molecules Cell Line Cells Cellular biology Co-Immunoprecipitations DNA Binding Data Data Quality Development Dominant-Negative Mutation Dose Effector Cell Electrophoretic Mobility Shift Assay Elements Enzyme-Linked Immunosorbent Assay Flow Cytometry Fractalkine Functional disorder GRP gene Genes Glucocorticoid Receptor Glucocorticoids Goals Growth Factor Health Health Care Costs Hour Immune Inflammation Inflammatory Inflammatory Response Intercellular adhesion molecule 1 Interferons Knowledge Laboratories Lung diseases Mediating Mentors Molecular PP5 protein-serine-threonine phosphatase Pathogenesis Pathway interactions Patients Pharmacology Phase Phosphoric Monoester Hydrolases Phosphorylation Play Principal Investigator Property Protein Isoforms Protein Serine/Threonine Phosphatase Proteins Public Health RANTES RNA Interference Recruitment Activity Regulation Relative (related person)Reporter Research Research Personnel Resistance Role Serine Signal Transduction Smooth Muscle Myocytes Steroid Receptors Steroid Resistance Steroid therapy Steroids Techniques Technology Testing Therapeutic Threonine Time Tissues Transactivation Transfection Up-Regulation Western Blotting airway inflammation airway obstruction chemokine chromatin immunoprecipitation cytokine design experience glucocorticoid receptor beta glucocorticoid receptor-interacting protein 1 human TIF2 factor immunocytochemistry improved inhibitor/antagonist novel novel therapeutics nucleocytoplasmic transport overexpression programs respiratory smooth muscle transcription factor vector

项目摘要

Although steroids are highly effective in the control of asthma, some patients fail to respond even to high doses. Steroid resistance is a veritable health challenge due to the absence of therapeutic alternatives and a financial burden as steroid-resistant patients account for more than 50% of asthma-related healthcare costs. The awardee's research focuses on studying steroid resistance in airway smooth muscle (ASM), a tissue that is relevant for lung diseases. The short-term goal of the studies was achieved during the one-year mentored phase of the K99 award with the identification of some of the molecular mechanisms responsible for mediating IRF-1-induced inhibition of steroid function in ASM cells. Interestingly, decreasing IRF-1 levels restores only partially steroid responsiveness in cytokine-treated cells suggesting that pathways, other than IRF-1, could be involved in cytokine-induced steroid resistance and are the aims of, the studies under the ROD phase. Thus, the main goal of ROO award is to investigate the contribution of other inflammatory molecules in cytokine-induced steroid resistance with the ultimate objective to generate novel potential therapeutic options to treat steroid-resistant asthmatics. The central hypothesis of the ROO proposal is novel and states that pro-asthmatic cytokines impair steroid function in ASM cells through the coordinated activation of two IRF-1-independent pathways: (i) glucocorticoid receptor beta isoform (GRP), a steroid receptor beta isoform that can act as an inhibitor of GC actions (will be addressed in Aim 1), and (ii) Serine/threonine protein phosphatase 5 (PP5), shown to act as an inhibitor of steroid actions in different cell lines (will be addressed in Aim 2). These latter two aims of the present proposal will rely on multiple complementary approaches such as siRNA technology, transfection of reporter vectors as well as overexpression of constitutively active or dominant negative proteins, co-immunoprecipitation and co-locaiization techniques, chromatin immunoprecipitation and gel shift assays. Subsequently, this award will dramatically strenghten the applicant's independence, will broaden his expertise in molecular pharmacology and cell biology, and importantly will yield high quality data necessary for his projected R01 application.

尽管类固醇在控制哮喘方面非常有效，但一些患者甚至对高剂量的反应也没有反应。由于缺乏治疗性替代品，类固醇耐药性是一项名副其实的健康挑战，而且由于抗类固醇的患者的财务负担占与哮喘相关的医疗费用的50％以上。获奖者的研究重点是研究气道平滑肌（ASM）中的类固醇耐药性，这是一种与肺部疾病有关的组织。研究的短期目标是在K99奖励的一年指导阶段实现的，并鉴定了一些分子机制，这些分子机制负责介导IRF-1诱导的ASM细胞中类固醇功能的抑制。有趣的是，降低IRF-1水平仅恢复细胞因子处理的细胞中部分类固醇的反应能力，这表明除IRF-1以外的途径可能与细胞因子诱导的类固醇抗性有关，并且是在研究下的目的，并且是研究的目的。杆相。因此，ROO奖的主要目标是研究其他炎症分子在细胞因子诱导的类固醇耐药中的贡献，并具有产生新型的潜在治疗选择以治疗类固醇哮喘患者的最终目标。 The central hypothesis of the ROO proposal is novel and states that pro-asthmatic cytokines impair steroid function in ASM cells through the coordinated activation of two IRF-1-independent pathways: (i) glucocorticoid receptor beta isoform (GRP), a steroid receptor beta isoform that can act as an inhibitor of GC actions (will be addressed in Aim 1), and (ii) Serine/threonine蛋白质磷酸酶5（PP5），表现为在不同细胞系中的类固醇作用的抑制剂（将在AIM中解决 2）。本提案的后两个目标将依赖于多种互补方法，例如siRNA技术，转染记者向量以及过表达组成型活性或显性负蛋白，共免疫沉淀和共终止技术，染色质免疫沉淀和凝胶转移测定法。随后，该奖项将极大地影响申请人的独立性，将扩大他在分子药理学和细胞生物学方面的专业知识，重要的是，重要的是，他的预计R01应用所需的高质量数据。