Mechanism of inflammation-induced steroid resistance

炎症诱导类固醇抵抗的机制

• 批准号: 7486327
• 负责人: OMAR TLIBA
• 金额: $ 9万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-08-15 至 2008-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7486327
• 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; Development; Disease; Dominant-Negative Mutation; Dose; Effector Cell; Electrophoretic Mobility Shift Assay; Elements; Enzyme-Linked Immunosorbent Assay; Figs - dietary; Flow Cytometry; Fractalkine; Functional disorder; GRP gene; Genes; Glucocorticoid Receptor; Glucocorticoids; Goals; Growth Factor; Health; Health Care Costs; Hour; IRF1 gene; Immune; Inflammation; Inflammatory; Inflammatory Bowel Diseases; Inflammatory Response; Intercellular adhesion molecule 1; Interferons; Knowledge; Laboratories; Lead; Lung diseases; Mediating; Mentors; Mesenchymal; Molecular; PP5 protein-serine-threonine phosphatase; Pathogenesis; Pathway interactions; Patients; Pennsylvania; Pharmacology; Phase; Phosphoric Monoester Hydrolases; Phosphorylation; Play; Polymerase Chain Reaction; Property; Protein Isoforms; Protein Overexpression; 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; Steroids; Techniques; Technology; Testing; Therapeutic; Therapeutic Agents; Threonine; Time; Tissues; Transactivation; Transfection; Universities; Up-Regulation; Western Blotting; airway obstruction; chemokine; chromatin immunoprecipitation; cytokine; design; experience; glucocorticoid receptor beta; glucocorticoid receptor-interacting protein 1; human TIF2 factor; immunocytochemistry; improved; inhibitor/antagonist; insight; novel; novel therapeutics; nucleocytoplasmic transport; programs; respiratory smooth muscle; skills; therapeutic target; transcription factor; vector

DESCRIPTION (provided by applicant): 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 candidate's current research at the University of Pennsylvania focuses in studying steroid resistance in airway smooth muscle (ASM), a tissue that is relevant for lung diseases. The short term goal of the studies performed currently and during the mentored phase of this award (K99) is to delineate the molecular mechanisms that underlay the diminished efficacy of steroids. This study will be pursued during the independent phase of this award (R00) with the ultimate objective to develop new therapeutic options to treat steroid-resistant asthmatics. The candidate's long term goal is to establish his own independent research program and submit an R01 application. The central hypothesis of this proposal is novel and states that pro-asthmatic cytokines impair steroid function in ASM cells through the coordinated activation of three pathways: (i) GR beta, a steroid receptor beta isoform that can act as an inhibitor of GC actions (will be addressed in Aim 1), (ii) IRF-1, a transcription factor that regulates many inflammatory genes (will be addressed in Aim 2), and (iii) Serine/threonine protein phosphatase 5 (PP5), shown to act as an inhibitor of steroid actions in different cell lines (will be addressed in Aim 3). All three Aims of the present proposal will rely on multiple complementary approaches such as siRNA technology, transfection of reporter vectors as well as over-expression of constitutively active or dominant negative proteins, co-immunoprecipitation and co-localization techniques, chromatin immunoprecipitation and gel shift assays. Subsequently, this award will dramatically enhance the candidate's technical skills and will broaden his expertise in molecular pharmacology and cell biology. We believe that our proposal is relevant to public health since improving the knowledge about the factors that lead to steroid resistance will help design new therapeutic agents or alternatives to treat steroid-resistant asthmatics.

描述（由申请人提供）： 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。由于缺乏治疗替代品，因此类固醇耐药性是一项名副其实的健康挑战，并且由于类固醇耐药性患者占相关医疗费用的50％以上。候选人目前在宾夕法尼亚大学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。目前进行的研究的短期目标以及该奖项的指导阶段（K99）是描述降低类固醇功效降低的分子机制。这项研究将在该奖项的独立阶段（R00）中进行，其最终目标是开发新的治疗选择以治疗耐药哮喘患者。候选人的长期目标是建立自己的独立研究计划并提交R01申请。该提议的中心假设是新颖的，并指出，促侵细胞因子通过三种途径的协调激活损害了ASM细胞中类固醇的功能：（i）GR Beta，一种可以作为GC作用抑制剂的类固醇受体β同工型（I）将在AIM 1），（ii）IRF-1中解决，它是调节许多炎症基因的转录因子（将在AIM 2中解决），以及（iii）丝氨酸/苏氨酸蛋白磷酸酶5（PP5），显示为充当不同细胞系中类固醇作用的抑制剂（AIM 3将解决）。本提案的所有三个目标都将依赖于多种互补方法，例如siRNA技术，记者向量的转染以及组成型活性或显性负蛋白的过度表达，共免疫沉淀和共闭合化技术，染色质免疫沉淀和凝胶移位和凝胶转移测定。随后，该奖项将大大提高候选人的技术技能，并扩大他在分子药理学和细胞生物学方面的专业知识。我们认为，我们的建议与公共卫生有关，因为提高了导致类固醇耐药性的因素的知识将有助于设计新的治疗剂或替代药物以治疗耐药哮喘患者。