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）： 肺纤维化可能在职业接触二氧化硅和石棉后发生，或者在没有明确原因的情况下发生，如特发性肺纤维化（IPF），不幸的是，人们认为这些疾病没有有效的治疗方法。损伤导致纤维生长因子过度产生，例如 TGF-¿我的实验室令人兴奋的初步研究表明，PTP 的遗传缺陷会导致进行性纤维化。在小鼠模型中预防肺纤维化，而不改变急性炎症反应的发生或消退，这种特异性表明 PTP¿ ，一种具有可靶向细胞外结构域的跨膜 PTP，可能是纤维化的有希望的药物靶标，并强调了确定 PTP 如何发挥作用的重要性。我们还发现 PTP 会促进肺纤维化。增强 TGF-¿使用相互的骨髓嵌合体，我们确定了 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，目前可用的全球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导致了这种表型。对于描述 PTP 的分子机制至关重要。选择性地驱动肺部纤维化，我们与 PTP 作斗争。通过增强促纤维化 TGF-¿ 促进肺纤维化目标 1 是产生具有 PTP 的小鼠。在成纤维细胞或肺泡 (AT)II 型上皮细胞中选择性删除 我们购买了具有靶向 PTP 的 B6/N ES 细胞系。来自 EUCOMM 的等位基因，并使用它们生成嵌合小鼠，这些小鼠将交配生成具有 floxed PTP 的小鼠。等位基因 (PTP¿f/f) 生成成纤维细胞特异性的 PTP¿ ，我们将跨越PTP¿ f/f 小鼠与 Col1¿2-Cre 小鼠生成 ATII 特异性删除 PTP¿ ，我们将跨越PTP¿ f/f 小鼠与 Sftpc-Cre 小鼠我们将确定是否 PTP¿促进 TGF-¿ - 在培养的成纤维细胞和从这些小鼠中分离的 ATII 细胞中诱导促纤维化信号传导 目的 2 是确定 PTP 的重要性。使用细胞类型特异性基因靶向小鼠在肺成纤维细胞或 ATII 细胞中促进肺纤维化，我们将比较三种模型中肺纤维化的严重程度：(i) 博来霉素、(ii) 二氧化硅或 (iii) 腺病毒的气管内滴注。表达活性 TGF¿在 PTP 基因缺陷的小鼠中在成纤维细胞 (Col1¿2-Cre/PTP¿f/f) 或 ATII 细胞 (Sftpc-Cre/PTP¿f/f) 中，这种高影响力的方法将使我们能够确定 PTP 是否如此。控制 TGF-¿成纤维细胞或 ATII 细胞中依赖的促纤维化信号传导，也将提供 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来治疗无意中接触含二氧化硅粉尘等纤维形成剂的个人（建筑工人、急救人员），以预防进行性肺纤维化。