Targeting Foxm1 in pulmonary fibrosis

靶向 Foxm1 治疗肺纤维化

基本信息

批准号：
9151958
负责人：
Tanya Kalin
金额：
$ 31.2万
依托单位：
CINCINNATI CHILDRENS HOSP MED CTR
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-09-01 至 2020-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9151958
关键词：
Alveolar Attenuated Binding Sites Bleomycin Blocking Antibodies Boxing CCL2 gene CXCL5 gene Cells Chest Chronic Data Epithelial Epithelial Cells Event Family member Fibroblasts Fibrosis Foxes Gene Expression Gene Targeting Genes Genetic Genetic Transcription Goals Hamman-Rich syndrome Health Human Hyperplasia In Vitro Inflammation Mediators Inflammatory Injury Integrins Laboratories Lead Lesion Lung Lung Inflammation MMP2 gene MMP9 gene Mediating Mediator of activation protein Metalloproteases Modeling Molecular Mus Myelogenous Myeloid Cells Outcome Pathogenesis Patients Phenotype Play Production Proliferating Proteins Pulmonary Fibrosis Pulmonary Inflammation Radiation Radiation Pneumonitis Recruitment Activity Refractory Role Signal Pathway Signal Transduction Site-Directed Mutagenesis Stimulus Structure of parenchyma of lung Testing Tissues Transcript Transcriptional Regulation Transgenic Mice Transgenic Organisms Type II Epithelial Receptor Cell Western Blotting Winged Helix alveolar epithelium alveolar type II cell chemokine efficacy testing fibrogenesis gain of function improved indium-bleomycin inhibitor/antagonist irradiation loss of function lung injury lung regeneration macrophage member mouse model new therapeutic target novel novel strategies novel therapeutic intervention osteopontin overexpression prevent programs promoter repaired response small molecule transcription factor transcriptome sequencing

项目摘要

PROJECT SUMMARY. Existing treatments for pulmonary fibrosis have not significantly improved survival; there is a critical need for new approaches. A combination of environmental and genetic factors creates an alveolar epithelium that is susceptible to injury. Deregulated repair of damaged tissue leads to the hyperplastic and proliferating alveolar type II cells (AECII) – the so called “re-programmed” AECII, which are main pathological features in the lungs of patients with fibrosis. Re-programmed AECIIs play a key role in the pathogenesis of pulmonary fibrosis, producing TGF- and pro-inflammatory mediators leading to activation of lung fibroblasts and recruitment of macrophages that further deregulate repair. Our long-term goal is to dissect transcriptional regulation of pulmonary fibrosis. We recently identified a novel pro-fibrotic regulator, Foxm1, a member of the family of Forkhead Box (Fox) transcription factors. Our preliminary data demonstrated that Foxm1 is induced in AECII within fibrotic lesions, but not in normal alveolar region of human and mouse lungs, indicating that Foxm1 can be a marker of re-programmed AECIIs. Transgenic expression of activated Foxm1 transcript in mouse AECII exacerbated radiation-induced pneumonitis and caused severe pulmonary fibrosis. Conditional deletion of Foxm1 from AECII attenuated radiation-induced lung fibrosis. While these data demonstrate that AECII promote pulmonary fibrosis through Foxm1-mediated events, the downstream signaling pathways regulated by Foxm1 in AECII remain to be identified (objective). We will test hypothesis that reprogrammed hyperplastic AECII promote pulmonary fibrosis through Foxm1-mediated activation of fibroblasts and recruitment of myeloid inflammatory cells into fibrotic lesions. By combining genetic and pharmacological approaches in radiation- and bleomycin-induced mouse models of lung fibrosis, the proposed studies will identify molecular mechanisms regulated by Foxm1 in AECII, and determine contribution of hyperplastic Foxm1-positive AECII to ling fibrogenesis. In Aim 1, we will determine the role of Foxm1-positive AECII in activation of fibroblasts. Using transgenic mice with Foxm1 gain-of-function and loss-of-function, and single cell RNA-seq analysis of AECII isolated from human IPF lungs, we will identify Foxm1 target genes. Since our preliminary data show increased expression of pro-fibrotic and inflammatory mediators in Foxm1-overexpressing AECII, we will examine whether Foxm1 activates transcription of Osteopontin and TGFβ1 genes in AECII, and stimulates activation of latent TGFβ1 protein, causing activation of lung fibroblasts. In Aim 2, we will examine whether Foxm1 activates CCL2 and CXCL5 genes in AECII, leading to recruitment of myeloid inflammatory cells into fibrotic lungs. In Aim 3, we will use novel small molecule Foxm1 inhibitor recently discovered in my laboratory to inhibit AECII re-programming, lung inflammation and fibrotic remodeling in murine models of pulmonary fibrosis. Completion of our studies will (1) identify novel molecular mechanisms whereby Foxm1 induces pulmonary fibrosis, and (2) test efficacy of novel Foxm1 inhibitors in lung fibrosis.

项目摘要。现有的肺纤维化治疗方法并未显着提高生存率；因此迫切需要环境和遗传因素的结合创造了一种肺泡上皮细胞受损组织的修复失调会导致肺泡增生和增殖。 II 型细胞 (AECII) – 所谓的“重新编程”AECII，是肺部的主要病理特征重新编程的 AECIIs 在肺纤维化的发病机制中发挥着关键作用，产生 TGF-β 和促炎介质，从而激活肺成纤维细胞并招募我们的长期目标是剖析巨噬细胞的转录调控。我们最近发现了一种新型促纤维化调节因子 Foxm1，它是肺纤维化家族的成员。我们的初步数据表明 Foxm1 在 AECII 中被诱导。在纤维化病变内，但不在人和小鼠肺的正常肺泡区域中，表明 Foxm1 可以是小鼠 AECII 中激活的 Foxm1 转录物转基因表达的标记。加剧了放射性肺炎并导致严重的肺纤维化。 AECII 中的 Foxm1 减弱了辐射引起的肺纤维化，而这些数据表明 AECII 通过 Foxm1 介导的事件促进肺纤维化，下游信号通路受 AECII 中的 Foxm1 仍有待鉴定（客观），我们将检验重编程增生的假设。 AECII 通过 Foxm1 介导的成纤维细胞激活和募集促进肺纤维化通过结合遗传和药理学方法，将骨髓炎症细胞转化为纤维化病变。在辐射和博来霉素诱导的小鼠肺纤维化模型中，拟议的研究将确定分子 Foxm1 在 AECII 中调节的机制，并确定增生性 Foxm1 阳性 AECII 对 AECII 的贡献在目标 1 中，我们将确定 Foxm1 阳性 AECII 在成纤维细胞激活中的作用。 Foxm1功能获得和功能丧失的转基因小鼠，以及AECII的单细胞RNA-seq分析从人类 IPF 肺部分离，我们将鉴定 Foxm1 靶基因，因为我们的初步数据显示增加。 Foxm1 过表达 AECII 中促纤维化和炎症介质的表达，我们将检查 Foxm1是否激活AECII中骨桥蛋白和TGFβ1基因的转录，并刺激潜在的 TGFβ1 蛋白，导致肺成纤维细胞激活在目标 2 中，我们将检查 Foxm1 是否激活。 AECII 中的 CCL2 和 CXCL5 基因，导致骨髓炎症细胞募集到纤维化的肺部。目标3，我们将使用我实验室最近发现的新型小分子Foxm1抑制剂来抑制AECII 肺纤维化小鼠模型中的重编程、肺部炎症和纤维化重塑。完成我们的研究将 (1) 确定 Foxm1 诱导肺损伤的新分子机制 (2) 测试新型 Foxm1 抑制剂对肺纤维化的疗效。