Mesenchymal Cell Dysfunction in Fibroproliferative Lung Disease

纤维增生性肺病中的间充质细胞功能障碍

基本信息

批准号：
10198011
负责人：
Paul Wesley Noble
金额：
$ 51万
依托单位：
CEDARS-SINAI MEDICAL CENTER
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-08-01 至 2023-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10198011
关键词：
Achievement Adhesions Adult Allografting Animal Model Apoptosis Attention Autoimmunity Basement membrane CD44 gene Cell Adhesion Cells Chronic Collaborations Detection Dissection Epithelial Epithelial Cells Extracellular Matrix Fibroblasts Fibrosis Focal Adhesion Kinase 1 Focal Adhesions Functional disorder Funding Genes Genetic Goals HAS2 gene Human Immune Immune Evasion Immune system Immunologic Receptors Immunologic Surveillance In Vitro Inflammation Ligands Light Lung Lung Transplantation Lung diseases Malignant Neoplasms Mediating Mesenchymal Mesenchymal Stem Cells Molecular Morbidity - disease rate Mus Myofibroblast PD-1 pathway PD-1/PD-L1 Pathogenesis Pathologic Pathway interactions Patients Pharmaceutical Preparations Phenotype Play Progressive Disease Pulmonary Fibrosis Regulation Role Sampling Source System TP53 gene Testing Tissues Tyrosine Kinase Inhibitor alveolar destruction alveolar epithelium beta-arrestin cancer cell effective therapy idiopathic pulmonary fibrosis immune checkpoint in vivo injured loss of function lung allograft migration mortality new therapeutic target novel novel strategies prevent programmed cell death ligand 1 programmed cell death protein 1 rho single cell analysis stem cell division transcriptome sequencing

项目摘要

Abstract Progressive fibrosis is a cause of major morbidity and mortality as best exemplified by idiopathic pulmonary fibrosis (IPF). The molecular mechanisms that control unremitting lung fibrosis remain poorly understood. During the funding period of this PO1, we identified an invasive fibroblast phenotype that appears to be an important feature of unremitting pulmonary fibrosis. We have identified invasive fibroblasts that contribute to severe lung fibrosis in mouse and human, and the invasive phenotype requires HAS2, CD44, and beta-arrestins. Fibroblast lineage-tracing studies identified that T box gene 4 (Tbx4)-lineage mesenchymal progenitors are the predominant source of myofibroblasts in injured adult murine lung. To further analyze the genetic regulation of fibroblast invasion, we performed RNA-seq analysis of a set of invasive and noninvasive fibroblast samples from IPF patients. Unexpectedly, we found that several immune molecules including both checkpoint PD1 ligands, PD-L1 (CD274) and PD-L2 (CD273), are significantly upregulated preferentially on invasive human fibroblasts. The Programmed Death-1 (PD-1, CD279) pathway is a homeostatic mechanism of the immune system that prevents autoimmunity and uncontrolled inflammation, and it is also used by cancer cells to escape from immune surveillance. This is an intriguing and novel discovery, particularly in light of the FDA approval of the multi- tyrosine kinase inhibitor nintedanib for IPF, a drug that was initially developed for cancer. A paradigm has evolved that the pathobiology of IPF is more similar to cancer than to chronic inflammation. However, the role of PD- 1/PD-L1 in lung fibrosis is unknown. Our preliminary studies revealed that PD-L1 regulates fibroblast adhesion, migration and invasiveness. We found that the focal adhesion components including focal adhesion kinase (FAK) and Rho/ROCK are significantly up-regulated. More recently, in collaboration with Dr. John Belperio at UCLA, PI on Project 3, we have also identified invasive fibroblasts in patients suffering from chronic lung allograft dysfunction (CLAD), which is the major cause of morbidity and mortality following lung transplantation. We propose to perform a similar approach in these cells with the hope of identifying new targets for therapy of CLAD. We have begun to examine the regulation of PD-L1 and found that PD-L1 was down-regulated by p53 in lung fibroblasts. In contrast to the immune checkpoint genes, p53 is markedly down-regulated in invasive fibroblasts, similar to that in cancer cells. Much attention has been paid to epithelial p53 expression and function in fibrosis, but the role of p53 in fibroblasts has not been thoroughly investigated. It appears that p53 may have a role not only in preventing epithelial apoptosis, but also in fibroblast invasiveness, thereby providing two distinct mechanisms in the pathogenesis of pulmonary fibrosis. Taken together, we hypothesize that loss of p53 up- regulates immune checkpoint genes in lung fibroblasts, leading to the activation of PD-1 ligand-mediated evasion of immune detection, focal adhesion and cell invasiveness.

抽象的进行性纤维化是特发性肺的最佳体现的主要发病率和死亡率的原因纤维化（IPF）。控制无ung纤维化的分子机制仍然鲜为人知。期间在此PO1的资金期间，我们确定了一种侵入性成纤维细胞表型，似乎很重要肺纤维化不懈的特征。我们已经确定了造成严重肺的侵入性成纤维细胞小鼠和人类的纤维化以及侵入性表型需要HAS2，CD44和β-art蛋白。成纤维细胞谱系追踪研究确定t盒基因4（TBX4） - 层间充质祖细胞是受伤的成年鼠肺肌纤维细胞的主要来源。进一步分析成纤维细胞入侵，我们对一组侵入性和非侵入性成纤维细胞样品进行了RNA-Seq分析 IPF患者。出乎意料的是，我们发现包括两个检查点PD1配体在内的几种免疫分子， PD-L1（CD274）和PD-L2（CD273）在侵袭性人成纤维细胞上优先上调。编程的死亡1（PD-1，CD279）途径是免疫系统的稳态机制，防止自身免疫性和不受控制的炎症，并且癌细胞也使用它来逃脱免疫监视。这是一个有趣而新颖的发现，尤其是鉴于FDA的批准酪氨酸激酶抑制剂Nintedanib用于IPF，这是一种最初用于癌症的药物。范式发展了 IPF的病理生物学与癌症更相似，而不是慢性炎症。但是，PD的作用肺纤维化中的1/PD-L1尚不清楚。我们的初步研究表明，PD-L1调节成纤维细胞粘附，移民和侵入性。我们发现焦点粘附成分在内 Rho/Rock明显上调。最近，与加州大学洛杉矶分校的John Belperio博士合作 PI在项目3上，我们还确定了患有慢性肺同种异体患者的侵入性成纤维细胞功能障碍（外壳），这是肺移植后发病和死亡率的主要原因。我们建议在这些细胞中执行类似的方法，以期识别出新靶标的外壳。我们已经开始检查PD-L1的调节，发现PD-L1在肺中被p53下调成纤维细胞。与免疫检查点基因相反，p53在侵入性成纤维细胞中明显下调，与癌细胞相似。对上皮p53表达和纤维化功能的关注很大，但是尚未彻底研究p53在成纤维细胞中的作用。看来p53可能没有角色仅在防止上皮细胞凋亡，但还具有成纤维细胞的侵袭性，从而提供了两个不同的肺纤维化发病机理的机制。综上所述，我们假设p53上的损失 - 调节肺成纤维细胞中的免疫检查点基因，导致PD-1配体介导的逃避激活免疫检测，局灶性粘附和细胞侵袭性。