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，进行性且通常是致命的肺部疾病 病因，以肺成纤维细胞激活和增殖为特征，细胞外过量沉积 矩阵（ECM）和普通肺建筑的破坏。 IPF患者的长期生存率很差， 5年生存率仅为20％。从历史上看，抗炎剂一直是治疗的主要手段，但是 事实证明，在降低疾病的严重程度或进展方面无效。因此，有 鉴定出新的可药物靶标，以开发有效的新疗法以识别用于治疗的新疗法的需求 IPF患者。由于肺成纤维细胞是IPF中的主要效应细胞，因此可以鉴定出新的可药物 控制IPF肺成纤维细胞生物学的分子或途径可能会导致更具体，更有效 IPF的治疗干预。在此提案中，我们试图确定是否系列/苏氨酸蛋白是 激酶D（PKD）家族激酶在IPF成纤维细胞激活，ECM的产生和表达中起关键作用 纤维纤维生长因子受体。 PKD家族激酶包括PKD1，PKD2和PKD3。在这里，我们发现 第一次在IPF的标志性病变的成纤维细胞灶中很容易检测到所有PKD同工型。我们 进一步表明，PKD1和PKD2在主要IPF成纤维细胞中始终激活，但不能控制 正常的人肺成纤维细胞。此外，PKD1和PKD2可以通过纤维化生长进一步激活 主要IPF成纤维细胞中的因素。有趣的是，PKD抑制明显下调了蛋白质水平 Procollagen和纤连蛋白没有显着影响其IPF成纤维细胞中的mRNA水平。而PKD 抑制明显抑制了mRNA和蛋白质水平的纤维化血小板衍生生长因子 成纤维细胞中的接收器 - （pDGFR¿）。根据这些新颖的发现，此重新提交R21建议是 旨在测试PKD家族激酶可能在IPF成纤维细胞生物学中起关键作用的假设， 包括成纤维细胞激活，ECM产生和纤维化生长因子受体的表达 可能会提供新的可吸毒目标，以阻止IPF进展。该重新提议的具体目的 为：目标1）表征构成性和纤维性激动剂引起的PKD家族激酶的激活 主要IPF成纤维细胞；目标2）确定PKD家族激酶的作用和机制 主要IPF成纤维细胞中的纤维化基质的产生；目标3）确定PKD家族激酶的作用 以及纤维细胞表达的纤维细胞表达的机制。我们预料到了 建议将识别PKD家族激酶是涉及肺成纤维细胞生物学的新颖和重要调节剂 在IPF的发病机理和进展中，靶向抑制PKD家族激酶或特定的PKD 同工型可以为IPF患者提供新颖有效的热干预。