Targeting PTPalpha to Prevent Lung Fibrosis

靶向 PTPα 预防肺纤维化

• 批准号: 8682312
• 负责人: Gregory Paul Downey
• 金额: $ 23.78万
• 依托单位: NATIONAL JEWISH HEALTH
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-05-05 至 2016-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8682312
• 关键词: Acute; Adenoviruses; Alleles; Alveolar; Antibodies; Asbestos; Automobile Driving; Biological; Bleomycin; Bone Marrow; Cells; Chimera organism; Cicatrix; Communities; Complex; Data; Disease; Drug Targeting; Dust; ES Cell Line; Environmental Exposure; Enzymes; Epithelial Cells; Exposure to; Extracellular Domain; Fibroblasts; Fibrosis; Fire - disasters; Gene Targeting; Genes; Genetic; Goals; Growth Factor; Hamman-Rich syndrome; Histologic; Human; In Vitro; Individual; Inflammatory Response; Integral Membrane Protein; Internal Ribosome Entry Site; Knock-out; Knowledge; Laboratories; LacZ Genes; Lung; Modeling; Molecular; Mus; Occupational Exposure; Partner in relationship; Pathway interactions; Phenotype; Phosphoric Monoester Hydrolases; Process; Production; Protein Tyrosine Phosphatase; Pulmonary Fibrosis; Resolution; Resources; Role; Severities; Signal Pathway; Signal Transduction; Silicon Dioxide; Smoke; Specificity; Therapeutic; Transgenic Mice; Western Blotting; alveolar type II cell; cell type; effective therapy; embryonic stem cell; emergency service responder; fibrogenesis; lung injury; macrophage; prevent; promoter; public health relevance; recombinase; response

DESCRIPTION (provided by applicant): Pulmonary fibrosis may occur after occupational exposure to silica and asbestos or without an identifiable cause, as in Idiopathic Pulmonary Fibrosis (IPF). Unfortunately, there is no effective therapy for these disorders. It is thought tha nonresolving lung injury results in excessive production of fibrogenic growth factors, such as TGF-¿, which drive progressive fibrosis. Exciting preliminary studies from my laboratory have revealed that genetic deficiency of PTP¿ prevents pulmonary fibrosis in a murine model without altering the genesis or resolution of the acute inflammatory response. This specificity suggests that PTP¿, a transmembrane PTP with a targetable extracellular domain, could be a promising drug target for fibrosis and highlights the importance of determining how PTP¿ promotes lung fibrosis. We have also discovered that PTP¿ enhances TGF-¿ responsiveness in fibrogenic signaling pathways in the lung. Using reciprocal bone marrow chimeras, we determined that PTP¿-/- lung resident cells and not bone marrow-derived cells, confer the protective phenotype. However, the limitations imposed by the currently available globally gene-deficient mice prevent us from determining which lung resident cells are responsible for this phenotype. This information is critical to delineate the molecular mechanisms by which PTP¿ selectively drives fibrosis in the lung. We hypothesize that PTP¿ promotes pulmonary fibrosis by augmenting profibrotic TGF-¿ signals in lung fibroblasts. Aim 1 is to generate mice in which PTP¿ is selectively deleted in either fibroblasts or alveolar type (AT)II epithelial cells. We have purchased B6/N ES cell lines with a targeted PTP¿ allele from EUCOMM and used these to generate chimeric mice that will be mated to generate mice with a floxed PTP¿ allele (PTP¿f/f). To generate a fibroblast-specific deletion of PTP¿, we will cross PTP¿f/f mice with Col1¿2-Cre mice. To generate ATII-specific deletion of PTP¿, we will cross PTP¿f/f mice with Sftpc-Cre mice. We will determine whether PTP¿ promotes TGF-¿-induced profibrotic signaling in cultured fibroblasts and ATII cells isolated from these mice. Aim 2 is to determine the importance of PTP¿ in lung fibroblasts or ATII cells in promoting pulmonary fibrosis using cell type-specific gene-targeted mice. We will compare the severity of pulmonary fibrosis in three models: intratracheal instillation of (i) bleomycin, (ii) silica, or (iii) adenovirus expressing active TGF¿ in mice that are genetically deficient in PTP¿ in either fibroblasts (Col1¿2-Cre/PTP¿f/f) or ATII cells (Sftpc-Cre/PTP¿f/f). This high impact approach will enable us to determine whether PTP¿ controls TGF-¿ dependent profibrotic signaling in fibroblasts or ATII cells and will also provide a valuable resource for the community. Knowledge gained from these studies will ultimately be used to develop pharmacological or biological approaches to treat individuals (construction workers, first responders) who are inadvertently exposed to fibrogenic agents such as silica-containing dust to prevent progressive pulmonary fibrosis.

描述（由申请人提供）：肺纤维化可能发生在职业暴露于二氧化硅和石棉后，或没有可识别的原因，如特发性肺纤维化（IPF）。不幸的是，没有有效的治疗这些疾病。据认为，未解决的肺损伤导致过度生产的纤维化生长因子，如TGF-β，这驱动进行性纤维化。我实验室令人兴奋的初步研究表明，PTP的遗传缺陷可以预防小鼠模型中的肺纤维化，而不会改变急性炎症反应的发生或消退。这种特异性表明，PTP是一种具有靶向胞外结构域的跨膜PTP，可能是一种有前途的纤维化药物靶标，并强调了确定PTP如何促进肺纤维化的重要性。我们还发现PTP <$增强肺纤维化信号通路中TGF-<$的反应性。使用骨髓嵌合体，我们确定PTP-/-肺驻留细胞，而不是骨髓来源的细胞，赋予保护性表型。然而，目前全球可用的基因缺陷小鼠所施加的限制使我们无法确定哪些肺驻留细胞负责这种表型。这些信息对于描述PTP选择性驱动肺纤维化的分子机制至关重要。我们假设PTP <$通过增加肺成纤维细胞中促纤维化TGF-<$信号促进肺纤维化。目的1是产生在成纤维细胞或肺泡II型（AT）上皮细胞中选择性缺失PTP的小鼠。我们从EUCOMM购买了具有靶向PTP <$等位基因的B6/N ES细胞系，并使用这些细胞系产生嵌合小鼠，这些嵌合小鼠将交配产生具有floxed PTP <$等位基因（PTP <$f/f）的小鼠。为了产生成纤维细胞特异性的PTP <$缺失，我们将PTP <$f/f小鼠与Col 1 <$2-Cre小鼠杂交。为了产生ATII特异性的PTP ú缺失，我们将PTP ú f/f小鼠与Sftpc-Cre小鼠杂交。我们将确定PTP <$是否促进从这些小鼠分离的培养成纤维细胞和ATII细胞中TGF-<$诱导的促纤维化信号传导。目的2是使用细胞类型特异性基因靶向小鼠来确定肺成纤维细胞或ATII细胞中PTP <$在促进肺纤维化中的重要性。我们将在三种模型中比较肺纤维化的严重程度：在成纤维细胞（Col 1 <$2-Cre/PTP <$f/f）或ATII细胞（Sftpc-Cre/PTP <$f/f）中PTP <$遗传缺陷的小鼠中，气管内滴注（i）博莱霉素，（ii）二氧化硅或（iii）表达活性TGF <$的腺病毒。这种高影响力的方法将使我们能够确定PTP是否控制成纤维细胞或ATII细胞中的TGF-β依赖性促纤维化信号传导，并且还将提供 社区的宝贵资源。从这些研究中获得的知识最终将用于开发药理学或生物学方法，以治疗无意中暴露于纤维化因子（如含二氧化硅粉尘）的个体（建筑工人，第一反应者），以预防进行性肺纤维化。