Targeting Lipogenic and Angiogenic Mediators in Pulmonary Fibrosis

摘要：由于肺纤维化(PF)的诊断和治疗方式都面临重大挑战， 重要的是继续寻找新的和可行的药物靶点，以对抗与 疾病。我们建议结合使用传统的分子生物学技术和高通量 脂质组学和蛋白质组学方法识别新的蛋白质和脂肪生物标记物，并研究其 对爱国阵线的贡献。长期目标是开发一种针对PF的有效治疗策略，而这项研究 与国家心肺血液研究所的使命直接相关。《公约》的总体目标 建议的研究是确定新的生物标志物，并研究脂肪生成和血管生成之间的相互作用 可能成为PF治疗靶点的介体。最近的证据表明，脂肪在人体内具有重要作用 在PF中的中介，但对疾病进展过程中总体脂质成分的放松调节知之甚少。 初步数据表明，脂肪酸合成酶(FASN)是一种重要的脂肪生成酶 与肺损伤相关，在博莱霉素(BLM)诱导的肺纤维化中。AIM 1旨在研究FASN的作用 在PF的发病机制中，确定关键的FASN相关的脂质和蛋白质伙伴。初步蛋白质组学 博莱霉素对小鼠肺组织磷脂酰肌醇-3-激酶/蛋白激酶B的鉴定 和WNT(无翼整合-1)/β(β)-连环蛋白信号通路作为两个最重要的纤维化信号通路 涉及到的路径。PI3K/Akt和Wnt/β-连环蛋白通路已被证明可增强纤维化反应， 我们已经报道了血管内皮生长因子(VEGF)的关键作用，它是血管内皮生长因子的直接靶点 PI3K/Akt通路，在调节纤维化中的作用。已知FASN在其他肺部疾病中调节血管内皮生长因子，如 哮喘，并在乳腺癌和前列腺癌中激活Wnt/β-catenin信号。我们假设FASN 通过调节PI3K/AKT&GT、血管内皮生长因子和WNT/β-连环蛋白通路发挥其在PF中的作用。AIM 2的设计目的是 探讨Wnt/β-catenin和PI3K/Akt&gt；血管内皮生长因子信号在FASN调控中的作用 博莱曼诱导的PF。我们正在进行的体内研究表明，FASN抑制剂奥利司他显著抑制了 博莱曼诱导的PF。此外，FASN、血管内皮生长因子和WNT/β-连环蛋白之间也出现了很强的相互作用 PF中的信令。AIM 3旨在评估奥利司他等抗FASN药物是否介导了肺纤维化 通过调节PI3K/AKT和GT；血管内皮生长因子和WNT/β-连环蛋白信号通路，并研究其潜在的作用 奥利司他与PI3K/Akt&gt、血管内皮生长因子和Wnt/β-连环蛋白抑制剂联合治疗方案的建立 小路。对这种串扰的研究和相关关键生物标志物的识别将对 增加对PF发病机制的分子机制的了解，将有助于 为这一致命疾病制定潜在的治疗和预防策略。

Abstract: Due to the major challenges in both diagnostic and therapeutic modalities in pulmonary fibrosis (PF), it is important to continue identifying novel and viable drug targets to combat pathogenesis associated with the disease. We propose to use a combination of traditional molecular biology techniques and high-throughput lipidomics and proteomics approaches to identify novel protein and lipid biomarkers, and investigate their contribution to PF. The long-term goal is to develop an effective therapeutic strategy against PF, and this study is directly relevant to the mission of National Heart, Lung and Blood Institute. The overall objective of the proposed study is to identify novel biomarkers and study the interplay between lipogenic and angiogenic mediators that may be targeted for treatment of PF. Recent evidence suggests an important role for lipid mediators in PF, but little is known about the deregulation of overall lipid composition in disease progression. Preliminary data suggests a critical role for fatty acid synthase (FASN), an important lipogenic enzyme associated with lung injury, in bleomycin (BLM)-induced PF. Aim 1 is designed to investigate the role of FASN in the pathogenesis of PF and identify key FASN-associated lipid and protein partners. Preliminary proteomic analysis of BLM-treated mouse lung tissue identified Phosphatidylinositol-3-kinase/Protein Kinase B (PI3K/Akt) and Wnt (Wingless Integration-1)/beta(β)-catenin signaling pathways as the two most important fibrotic pathways involved. PI3K/Akt and Wnt/β-catenin pathways have been shown to potentiate fibrotic response, and we have reported on the critical role of vascular endothelial growth factor (VEGF), a direct target of PI3K/Akt pathway, in regulating fibrosis. FASN is known to regulate VEGF in other lung diseases such as asthma, and activate Wnt/β-catenin signaling in breast and prostate cancers. We hypothesize that FASN exerts its effects in PF by regulating PI3K/Akt>VEGF and Wnt/β-catenin pathways. Aim 2 is designed to investigate the mechanistic effect of Wnt/β-catenin and PI3K/Akt>VEGF signaling on regulation of FASN in BLM-induced PF. Our ongoing in vivo studies demonstrates that FASN inhibitor Orlistat significantly inhibited BLM-induced PF. Furthermore, a strong interplay has emerged between FASN, VEGF and Wnt/β-catenin signaling in PF. Aim 3 is designed to evaluate whether anti-FASN agents such as Orlistat mediate lung fibrosis by regulating PI3K/Akt>VEGF and Wnt/β-catenin signaling pathways, and investigate the potential development of a co-therapy regimen involving Orlistat and inhibitors of the PI3K/Akt>VEGF and Wnt/β-catenin pathways. Investigation of this crosstalk and identification of related key biomarkers will be important for the increased understanding of the molecular mechanisms involved in the pathogenesis of PF and will facilitate development of potential therapeutic and preventive strategies for this fatal disease.