Human Cells and Tissues/Sheppard

项目总结（见说明）： 特发性肺纤维化（IPF）占间质性肺病病例的一半以上。 患有IPF的患者发展为肺功能的进行性丧失，并且平均在诊断后3年死亡。肺泡11型细胞在IPF肺中是异常的。然而，它们在疾病发病机制中的确切作用仍不明确。该计划项目资助研究了上皮细胞通过三个病理过程直接参与肺纤维化形成的新假设：ER应激，上皮整合素avB 6激活TGPB，以及上皮向间充质转化。考虑这个 假设，该转化计划项目资助的主要目标集中在开发靶向IPF肺泡11型细胞中这些病理过程的新药。 在支持计划项目赠款的总体目标。 人体组织核心将：1。提供用于处理、编目和储存从IPF患者和正常对照获得的人生物样品的集中资源，2.从正常和纤维化的人肺中获得高度纯化的肺泡II型细胞分离物，用于测试候选药物化合物，和3;提供集中的资源，用于比较分析从正常和IPF肺分离的人II型细胞、肺碎片和切片中的蛋白质和基因表达。 该核心将在诱导经历ER应激、avB 6介导的TGFB激活或EMT的细胞和组织上进行表达微阵列和miRNA阵列，在存在或不存在抑制这些途径的药物的情况下，以鉴定可以由每个项目表征的机械信息生物标志物。来自这个集中资源的细胞和组织将用于改进针对三种上皮细胞依赖性促纤维化途径中的每一种的候选药物。项目1检查在IPF 11型细胞（其表现出增加的UPR）和正常11型细胞（其UPR通过在衣霉素中培养而被激活）中操纵未折叠蛋白质应答的化合物。肺组织的片段将用于项目2中，以测试抑制上皮细胞依赖性TGFB通过整联蛋白avB 6活化的化合物。对于项目3，人类组织核心的领导者开发了使用FACS分选的新方法来分离高纯度的 来自正常和纤维化人肺的肺泡II型细胞群。这些细胞将用于EMT的新型体外模型中，以测试阻断TGPB信号传导和EMT的候选化合物。

PROJECT SUMMARY (See instructions): Idiopathic pulmonary fibrosis (IPF) accounts for more than half of the cases of interstitial lung disease. Patients suffering from IPF develop a progressive loss of lung function and die, on average, 3 years after diagnosis. Alveolar type 11 cells are abormal in IPF lungs. However, their precise role in disease pathogenesis remains undefined. This program project grant examines the novel hypothesis that epithelial cells directly participate in the formation of lung fibrosis via three pathologic processes: ER stress, activation of TGPB by the epithelial integrin avB6, and epithelial to mesenchymal transition. Considering this hypothesis, the primary objective of this translational program project grant centers on developing novel drugs that target these pathological processes in IPF alveolar type 11 cells. In support of the overall objectives of the program project grant. The Human tissue core will: 1. provide a centralized resource for processing, cataloging and storing human biological samples obtained from patients with IPF and normal controls, 2. obtain highly purified isolates of alveolar type II cells from normal and fibrotic human lungs for use in testing candidate drug compounds, and 3; provide a centralized resource forthe comparative analysis of protein and gene expression in human type II cells, lung fragments and slices isolated from normal and IPF lungs. The core will perform expression microarrays and miRNA arrays on cells and tissues induced to undergo ER stress, avB6-mediated TGFB activation or EMT in the presence or absence of drugs to inhibit these pathways, to identify mechanistically informative biomarkers that can be characterized by each project. The cells and tissues from this centralized resource will be used to refine candidate drugs targeting each of the three epithelial cell dependent profibrotic pathways. Project 1 to examine compounds manipulating the unfolded protein response both in IPF type 11 cells, which exhibit an increased UPR, and in normal type 11 cells whose UPR is activated by culture in tunicamycin. Fragments of lung tissue will be used in project 2 to test compounds inhibiting epithelial cell dependent TGFB activation by the integrin avB6. For project 3, the leader of the human tissue core developed novel methods using FACS sorting to isolate highly pure populations of alveolar type II cells from normal and fibrotic human lungs. These cells will be used in a novel in vitro model of EMT to test candidate compounds blocking TGPB signalling and EMT.