PKD Family Kinase Function and Signaling in Lung Fibroblasts

肺成纤维细胞中 PKD 家族激酶的功能和信号转导

• 批准号: 8204401
• 负责人: HUA TANG
• 金额: $ 17.63万
• 依托单位: UNIVERSITY OF TEXAS HLTH CTR AT TYLER
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-12-01 至 2013-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8204401
• 关键词: Affect; Agonist; Anti-Inflammatory Agents; Architecture; Biology; Cells; Chronic; Collagen; Deposition; Disease Progression; Effector Cell; Etiology; Extracellular Matrix; Family; Fibroblasts; Fibronectins; Growth Factor; Growth Factor Receptors; Hamman-Rich syndrome; Human; Lead; Lesion; Lung; Lung diseases; Messenger RNA; Molecular; Pathogenesis; Pathway interactions; Patients; Phosphotransferases; Platelet-Derived Growth Factor Receptor; Play; Procollagen; Production; Protein Isoforms; Proteins; Rest; Role; Serine; Severities; Signal Transduction; Survival Rate; Testing; Therapeutic Intervention; Threonine; Time; base; design; effective therapy; novel; protein kinase D; receptor expression; transcription factor

Idiopathic pulmonary fibrosis (IPF) is a chronic, progressive and usually fatal lung disease of unknown etiology, characterized by lung fibroblast activation and proliferation, excessive deposition of extracellular matrix (ECM), and destruction of the normal lung architecture. Long-term survival of IPF patients is poor, with a 5-year survival rate of only 20%. Historically, anti-inflammatory agents have been the mainstay of therapy, but have been proven to be ineffective in reducing either the severity or progression of the disease. Hence, there is a profound need for the identification of novel drugable targets to develop efficacious new therapies for treating IPF patients. Because lung fibroblasts are primary effector cells in IPF, the identification of novel drugable molecules or pathways that control IPF lung fibroblast biology may lead to more specific and efficacious therapeutic intervention in IPF. In this proposal, we seek to determine whether the serine/threonine protein kinase D (PKD) family kinases play critical roles in IPF fibroblast activation, production of ECM, and expression of profibrotic growth factor receptors. PKD family kinases include PKD1, PKD2 and PKD3. Herein, we found for the first time that all the PKD isoforms were readily detected in fibroblastic foci, the hallmark lesions of IPF. We further showed that PKD1 and PKD2 were constitutively activated in primary IPF fibroblasts but not in control normal human lung fibroblasts. Moreover, PKD1 and PKD2 could be further activated by profibrotic growth factors in primary IPF fibroblasts. Interestingly, PKD inhibition markedly downregulated the protein levels of procollagen and fibronectin without significantly affecting their mRNA levels in IPF fibroblasts. Whereas, PKD inhibition markedly suppressed both mRNA and protein levels of profibrotic platelet-derived growth factor receptor-¿ (PDGFR¿) in the fibroblasts. On the basis of these novel findings, this resubmission R21 proposal is designed to test the hypothesis that PKD family kinases may play critical roles in IPF fibroblast biology, including the fibroblast activation, ECM production and expression of profibrotic growth factor receptors, which may offer novel drugable targets for halting IPF progression. The Specific Aims of this resubmission proposal are: Aim 1) To characterize the constitutive and profibrotic agonist-induced activation of PKD family kinases in primary IPF fibroblasts; Aim 2) To determine the roles of PKD family kinases and the mechanisms in the production of profibrotic matrix in primary IPF fibroblasts; Aim 3) To determine the roles of PKD family kinases and the mechanisms in IPF fibroblast expression of profibrotic PDGF receptor-¿. We anticipate that this proposal will identify PKD family kinases as novel and important modulators of lung fibroblast biology involved in the pathogenesis and progression of IPF, therefore targeting to inhibit PKD family kinases or a specific PKD isoform may provide a novel and efficacious therapeutic intervention for IPF patients.

特发性肺纤维化 (IPF) 是一种慢性、进行性且通常致命的肺部疾病，病因不明 病因学，特点是肺成纤维细胞活化和增殖、细胞外沉积过多 IPF 患者的长期生存率较差。 5年生存率仅为20% 历史上，抗炎药物一直是主要治疗方法，但。 已被证明对于减轻疾病的严重程度或进展无效。 广泛需要识别新的药物靶点以开发有效的新疗法 由于肺成纤维细胞是 IPF 的主要效应细胞，因此需要鉴定新型药物。 控制 IPF 肺成纤维细胞生物学的分子或途径可能会导致更特异和有效的治疗 在本提案中，我们试图确定丝氨酸/苏氨酸蛋白是否参与 IPF 的治疗干预。 激酶 D (PKD) 家族激酶在 IPF 成纤维细胞激活、ECM 产生和表达中发挥关键作用 促纤维化生长因子受体激酶包括 PKD1、PKD2 和 PKD3。 首次在成纤维细胞病灶（IPF 的标志性病变）中轻松检测到所有 PKD 亚型。 进一步表明 PKD1 和 PKD2 在原代 IPF 成纤维细胞中被组成性激活，但在对照中未激活 此外，促纤维化生长可进一步激活 PKD1 和 PKD2。 原代 IPF 成纤维细胞中的 PKD 抑制显着下调了 PKD 的蛋白水平。 前胶原和纤连蛋白，而不显着影响 IPF 成纤维细胞中的 mRNA 水平。 抑制显着抑制促纤维化血小板衍生生长因子的 mRNA 和蛋白质水平 受体-¿ (PDGFR¿) 在这些新的发现的基础上，这个重新提交的 R21 提案是。 旨在检验 PKD 家族激酶可能在 IPF 成纤维细胞生物学中发挥关键作用的假设， 包括成纤维细胞活化、ECM 产生和促纤维化生长因子受体表达， 可能会提供新的药物靶标来阻止 IPF 进展。 该重新提交提案的具体目标。 目的 1) 表征 PKD 家族激酶的组成型和促纤维化激动剂诱导的激活 原代 IPF 成纤维细胞；目的 2) 确定 PKD 家族激酶的作用及其机制 原代 IPF 成纤维细胞中促纤维化基质的产生；目的 3) 确定 PKD 家族激酶的作用 以及 IPF 成纤维细胞表达促纤维化 PDGF 受体的机制 -¿ .我们预计这 该提案将确定 PKD 家族激酶作为肺成纤维细胞生物学的新颖且重要的调节剂 参与 IPF 的发病机制和进展，因此靶向抑制 PKD 家族激酶或特定的 PKD 同工型可能为 IPF 患者提供一种新颖有效的治疗干预方法。