Focal Adhesion Modulation of IL-1 Signaling: Importance in Pulmonary Fibrosis

IL-1 信号传导的局部粘附调节：在肺纤维化中的重要性

• 批准号: 8089546
• 负责人: Gregory Paul Downey
• 金额: $ 39万
• 依托单位: NATIONAL JEWISH HEALTH
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-07-08 至 2013-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8089546
• 关键词: Address; Animal Model; Anti-Inflammatory Agents; Attenuated; Binding; Biochemical; Biological Assay; Biopsy Specimen; Bleomycin; Cell Line; Cells; Chronic; Cicatrix; Co-Immunoprecipitations; Collagen; Complex; Disease; Embryo; Endoplasmic Reticulum; Environmental Pollutants; Epithelium; Event; Extracellular Matrix; Extracellular Matrix Degradation; Extracellular Matrix Proteins; Extracellular Signal Regulated Kinases; Fibroblasts; Fibronectins; Fibrosis; Focal Adhesions; Genetic Polymorphism; Hamman-Rich syndrome; Host Defense; Human; Immunohistochemistry; In Vitro; Inflammatory; Injury; Integrins; Interleukin-1; Interleukin-1 Receptors; Interstitial Collagenase; Lung; Lung Inflammation; MMP3 gene; Matrix Metalloproteinases; Measures; Mediating; Mediator of activation protein; Membrane; Messenger RNA; Mitogen-Activated Protein Kinases; Modeling; Molecular; Mus; Mutation; Organ; Outcome; PTK2 gene; PTPN11 gene; Pathogenesis; Pathway interactions; Patients; Peptide Hydrolases; Peptides; Phosphorylation; Pneumonia; Process; Production; Protein Dephosphorylation; Protein Tyrosine Phosphatase; Proteins; Pulmonary Fibrosis; Rattus; Recombinant Proteins; Regulation; Reporting; Respiratory physiology; Risk; Role; Sampling; Severities; Signal Pathway; Signal Transduction; Signaling Molecule; Small Interfering RNA; Stromelysin 1; Structure of parenchyma of lung; Therapeutic; Tissues; Tyrosine Phosphorylation; Virus Diseases; alveolar destruction; base; cytokine; design; effective therapy; in vitro Model; in vivo; inhibitor/antagonist; interstitial; knock-down; laser capture microdissection; link protein; mutant; receptor; reconstitution; repaired; response; scaffold; small molecule; treatment strategy; Address; Animal Model; Anti-Inflammatory Agents; Attenuated; Binding; Biochemical; Biological Assay; Biopsy Specimen; Bleomycin; Cell Line; Cells; Chronic; Cicatrix; Co-Immunoprecipitations; Collagen; Complex; Disease; Embryo; Endoplasmic Reticulum; Environmental Pollutants; Epithelium; Event; Extracellular Matrix; Extracellular Matrix Degradation; Extracellular Matrix Proteins; Extracellular Signal Regulated Kinases; Fibroblasts; Fibronectins; Fibrosis; Focal Adhesions; Genetic Polymorphism; Hamman-Rich syndrome; Host Defense; Human; Immunohistochemistry; In Vitro; Inflammatory; Injury; Integrins; Interleukin-1; Interleukin-1 Receptors; Interstitial Collagenase; Lung; Lung Inflammation; MMP3 gene; Matrix Metalloproteinases; Measures; Mediating; Mediator of activation protein; Membrane; Messenger RNA; Mitogen-Activated Protein Kinases; Modeling; Molecular; Mus; Mutation; Organ; Outcome; PTK2 gene; PTPN11 gene; Pathogenesis; Pathway interactions; Patients; Peptide Hydrolases; Peptides; Phosphorylation; Pneumonia; Process; Production; Protein Dephosphorylation; Protein Tyrosine Phosphatase; Proteins; Pulmonary Fibrosis; Rattus; Recombinant Proteins; Regulation; Reporting; Respiratory physiology; Risk; Role; Sampling; Severities; Signal Pathway; Signal Transduction; Signaling Molecule; Small Interfering RNA; Stromelysin 1; Structure of parenchyma of lung; Therapeutic; Tissues; Tyrosine Phosphorylation; Virus Diseases; alveolar destruction; base; cytokine; design; effective therapy; in vitro Model; in vivo; inhibitor/antagonist; interstitial; knock-down; laser capture microdissection; link protein; mutant; receptor; reconstitution; repaired; response; scaffold; small molecule; treatment strategy

DESCRIPTION (provided by applicant): Idiopathic pulmonary fibrosis (IPF) is a progressive and frequently fatal disorder for which there are currently no effective treatment strategies. The current proposal focuses on Interleukin-1 (IL-1), a potent, pro-inflammatory cytokine that induces multiple signaling cascades in fibroblasts. These signals serve in host defense but, paradoxically, may contribute to inflammatory tissue injury and to fibrosis of the lung and other organs. IL-1 stimulates Ca2+ release and expression of multiple cytokines and inflammatory factors such as matrix metalloproteinases (MMPs) that drive extracellular matrix degradation via mitogen activated protein (MAP) kinase pathways. Currently, the mechanisms by which IL-1-induced signals are moderated or terminated are incompletely understood. Our studies to date have determined that in fibroblasts, IL-1-induced signaling requires focal adhesions (FA) and that IL-1 signals are dissipated by FA dispersing peptides that we have developed. Our recent studies indicate the importance of two protein tyrosine phosphatases (PTP), SHP- 2 and PTP(, in the regulation of IL-1-induced FA maturation and IL-1 induced signals. We discovered that SHP-2 mediates functional interactions between focal adhesions and the endoplasmic reticulum that are crucial for ER Ca2+ release; these interactions, together with focal adhesions, are central determinants of IL-1 signaling. Our hypothesis is that the PTP( in FA mediates maturation and remodeling of FA in response to IL- 1. In these multi-molecular signaling platforms, PTP( interacts with and dephosphorylates SHP-2, leading to recruitment and activation of additional signaling and scaffold molecules that are essential for Ca2+ release from the endoplasmic reticulum, signaling to ERK, and MMP-1 and 3 secretion. In Specific Aim 1 we will determine how PTP( regulates FA-dependent IL-1 signaling leading to ERK activation and MMP-1 and 3 secretion. We will use cultured human lung fibroblasts from normals and IPF lungs and murine fibroblasts from SHP-2 or PTP(-null embryos, reconstituted with wild type or mutant proteins, as in vitro models to study FA-restricted signaling. ERK activation and MMP1 and 3 release will be used as outcomes of IL-1 signaling to assess the impact of SHP-2 and PTP(. In Specific Aim 2, we will determine the role of PTP( in regulating IL-1 signaling through focal adhesions and the ER. We will examine the molecular determinants of PTP( and SHP-2 interactions and how they regulate the IP3 receptor and other FA-dependent signals leading to MMP expression. In Specific Aim 3, we will assess the roles of PTP( and MMP3 in vivo in a murine model of bleomycin-induced pulmonary fibrosis. We will also examine potential alterations in PTP( and SHP-2 expression and/or activation in samples of banked lung tissue from patients with pulmonary fibrosis. These samples will be used to determine if there are alterations in the expression of SHP2 and PTP( mRNA and protein in the lungs of these patients that are associated with severity of pulmonary fibrosis. As IL-1 is a critical mediator of chronic inflammatory conditions such as pulmonary fibrosis, elucidation of IL-1 signaling pathways is fundamental in the identification of specific targets for effective anti-inflammatory agents. This is exemplified by small molecule inhibitors of specific PTP and peptides designed to selectively interfere with pro-fibrotic pathways as potential therapeutics. PROJECT NARRATIVE: Pulmonary fibrosis (scarring of the lung) is a progressive and usually fatal disorder for which there is currently no effective therapy. It is currently believed that the scarring process is started by damage to the lining cells of the lung (epithelium), perhaps as the result of a viral infection or from environmental pollutants. We have discovered that Interleukin-1, a small molecule secreted by the cells lining the lung, strongly promotes pathways leading to scarring in the lung. We propose to characterize these pathways with the aim of identifying specific targets for therapies to curtail this devastating process.

描述（由申请人提供）：特发性肺纤维化（IPF）是一种进行性且经常致命的疾病，目前尚无有效的治疗策略。当前的建议着重于白介素-1（IL-1），这是一种有效的促炎细胞因子，可诱导成纤维细胞中的多个信号传导级联反应。这些信号用于宿主防御，但自相矛盾的是，可能导致炎症组织损伤以及肺部和其他器官的纤维化。 IL-1刺激CA2+释放和多种细胞因子和炎症因子的表达，例如基质金属蛋白酶（MMP），这些因子（MMP）通过有丝分裂原活化蛋白（MAP）激酶途径驱动细胞外基质降解。当前，尚不完全了解IL-1诱导的信号被调节或终止的机制。迄今为止，我们的研究确定，在成纤维细胞中，IL-1诱导的信号传导需要局灶性粘连（FA），并且IL-1信号通过我们开发的FA分散肽来消除IL-1信号。 Our recent studies indicate the importance of two protein tyrosine phosphatases (PTP), SHP- 2 and PTP(, in the regulation of IL-1-induced FA maturation and IL-1 induced signals. We discovered that SHP-2 mediates functional interactions between focal adhesions and the endoplasmic reticulum that are crucial for ER Ca2+ release; these interactions, together with focal adhesions, are central IL-1信号传导的决定因素。对ERK的信号传导，MMP-1和3个分泌。蛋白质，作为研究FA限制信号传导的体外模型。 ERK activation and MMP1 and 3 release will be used as outcomes of IL-1 signaling to assess the impact of SHP-2 and PTP(. In Specific Aim 2, we will determine the role of PTP( in regulating IL-1 signaling through focal adhesions and the ER. We will examine the molecular determinants of PTP( and SHP-2 interactions and how they regulate the IP3 receptor and other FA-dependent signals leading to MMP表达在特定的目标3中，我们将评估PTP的作用（在体内MMP3在博霉素诱导的肺纤维化模型中与肺纤维化的严重程度相关的这些患者的肺部蛋白质是慢性炎症状况的关键介体，例如肺纤维化，阐明IL-1信号传导途径是鉴定有效抗炎药的特定靶标的IL-1信号通路。这是特定PTP和肽的小分子抑制剂的例证，旨在选择性地干扰促纤维化途径作为潜在的疗法。项目叙事：肺纤维化（肺部疤痕）是一种进行性且通常是致命的疾病，目前尚无有效的治疗。目前认为，疤痕过程是由于病毒感染或环境污染物的结果而造成了对肺部内膜（上皮）的损害。我们已经发现，白细胞介素-1是肺内壁细胞分泌的小分子，强烈促进导致肺部疤痕的途径。我们建议对这些途径进行表征，目的是确定用于减少这种毁灭性过程的疗法的特定靶标。