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

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。我们最近的研究表明，两种蛋白酪氨酸磷酸酶（PTP），SHP-2和PTP的重要性（在调节IL-1诱导的FA成熟和IL-1诱导的信号中，我们发现SHP-2介导了SHP-2介导的功能相互作用焦点粘连和内质网络对于ER Ca2+释放至关重要； -1。在这些多分子信号平台中，PTP（与SHP-2相互作用并去磷酸化，从而导致募集和激活其他信号传导和脚手架分子，这对于Ca2+从内质网释放至关重要，这对于从内质网释放，信号传导到ERK和MMP------ 1和3分泌。在特定目标中，我们将确定PTP（调节FA依赖性IL-1信号传导，导致ERK激活以及MMP-1和3分泌。我们将使用培养的人类肺成纤维细胞。从SHP-2或PTP（-Null Embryos，用野生型或突变蛋白重构为体外模型，可以研究FA限制的信号传导。 ERK激活以及MMP1和3释放将用作IL-1信号传导的结果来评估SHP-2和PTP的影响（。在特定的AIM 2中，我们将确定PTP的作用（在通过局灶性调节IL-1信号方面粘连和ER。和mmp3在博来霉素诱导的肺纤维化的鼠模型中，我们还将检查PTP的潜在变化（以及SHP-2表达和/或激活的肺纤维化患者的样品。为了确定SHP2和PTP的表达是否有改变（这些患者的肺中的mRNA和蛋白质与肺纤维化严重程度相关。由于IL-1是慢性炎性疾病（例如肺纤维化）的关键介体，例如肺纤维化，因此IL-1信号传导途径的鉴定是有效抗炎剂的特定靶标。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。

项目成果

Interactions of the protein-tyrosine phosphatase-α with the focal adhesion targeting domain of focal adhesion kinase are involved in interleukin-1 signaling in fibroblasts.

蛋白酪氨酸磷酸酶-α 与粘着斑激酶的粘着斑靶向域的相互作用参与成纤维细胞中的白细胞介素-1 信号传导。

• DOI: 10.1074/jbc.m113.540294
• 发表时间: 2014
• 期刊: The Journal of biological chemistry
• 作者: Wang,Qin;Wang,Yongqiang;Fritz,Dominik;Rajshankar,Dhaarmini;Downey,GregoryP;McCulloch,ChristopherA
• 通讯作者: McCulloch,ChristopherA

Role of PTPα in the destruction of periodontal connective tissues.

• DOI: 10.1371/journal.pone.0070659
• 发表时间: 2013
• 期刊: PloS one
• 影响因子: 3.7
• 作者: Rajshankar D;Sima C;Wang Q;Goldberg SR;Kazembe M;Wang Y;Glogauer M;Downey GP;McCulloch CA
• 通讯作者: McCulloch CA

Reply: defining lung injury in animals.

答复：定义动物肺损伤。

• DOI: 10.1165/rcmb.2012-0074le
• 发表时间: 2013
• 期刊: American journal of respiratory cell and molecular biology
• 影响因子: 6.4
• 作者: Matute-Bello,Gustavo;Downey,GregoryP
• 通讯作者: Downey,GregoryP

Human neutrophil peptides and phagocytic deficiency in bronchiectatic lungs.

• DOI: 10.1164/rccm.200808-1250oc
• 发表时间: 2009-07
• 期刊: American journal of respiratory and critical care medicine
• 影响因子: 24.7
• 作者: Stefanos Voglis;K. Quinn;E. Tullis;Mingyao Liu;Melanie Henriques;C. Zubrinich;Ó. Peñuelas;Holman Chan;F. Silverman;V. Cherepanov;N. Orzech;A. Khine;A. Cantin;Arthur S Slutsky;G. Downey;Haibo Zhang

The Leucine-Rich Repeat Region of CARMIL1 Regulates IL-1-Mediated ERK Activation, MMP Expression, and Collagen Degradation.

• DOI: 10.1016/j.celrep.2020.107781
• 发表时间: 2020-06-30
• 期刊: Cell reports
• 影响因子: 8.8
• 作者: Wang Q;Notay K;Downey GP;McCulloch CA
• 通讯作者: McCulloch CA