Development of a High-Throughput Screen for Pre-therapeutic Discovery in Idiopathic Pulmonary Fibrosis

特发性肺纤维化治疗前发现的高通量筛选的开发

基本信息

批准号：
8864277
负责人：
Robert Damoiseaux
金额：
$ 33.33万
依托单位：
UNIVERSITY OF CALIFORNIA LOS ANGELES
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-03-01 至 2018-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8864277
关键词：
Apoptosis Atomic Force Microscopy Biochemical Biological Assay Biological Models Book Chapters Cell Culture Techniques Cell model Cells Cellular Morphology Cessation of life Chemicals Chronic Cicatrix Clinic Clinical Trials Collagen Complementary DNA Complex Data Collection Deposition Development Diagnosis Disease Disease model Employee Strikes Equipment Etiology Exhibits Extracellular Matrix Fibroblasts Fibrosis Foundations Gases Gene Expression Profiling Genes Goals Hamman-Rich syndrome Human Imaging Techniques In Vitro Libraries Lung Lung diseases Mesenchymal Modeling Molecular Myofibroblast National Institute of General Medical Sciences Pathogenesis Patients Pharmaceutical Preparations Phenotype Preclinical Drug Evaluation Prevention Process Rare Diseases Resistance Resource Sharing Resources Respiratory Failure Running Sampling Survival Rate System Testing Therapeutic Therapeutic Index Time Tissues Toxicity Tests Triage Validation Western Blotting Work Writing authority base cell type clinically relevant counterscreen disease phenotype disorder prevention drug development drug discovery drug efficacy functional genomics high throughput screening improved in vitro Model induced pluripotent stem cell interstitial killings novel prevent public health relevance screening small hairpin RNA small molecule stem cell biology stem cell technology tool

项目摘要

DESCRIPTION (provided by applicant): Idiopathic Pulmonary Fibrosis (IPF) is a scarring lung disease of unknown etiology killing over 40,000 patients each year in the US. The disease is characterized by aggregation of myofibroblasts in fibrotic foci and this lung remodeling results in poor gas exchange which is progressive and invariably fatal due to lack of treatment options. The understanding of the pathogenesis of IPF is very limited due to lack of good assay systems replicating the disease as well as lack of good model systems to study drug efficacy. We have generated a novel, robust, clinically relevant human model for IPF using induced pluripotent stem cells (iPSCs) from IPF patients. Specifically, this model lends itself to drug discovery as the patient derived iPSCs develop progressive fibrotic foci in the dish when redifferentiated to mesenchymal-type cells, thereby recapitulating the disease phenotype in a dish. Our Specific Aims are: 1. We will develop a high throughput screening (HTS) assay to identify small molecule probes for target discovery and validation, test already approved drugs for repurposing as IPF treatment and add new compounds for IPF drug development utilizing our "disease in a dish" model. Our in vitro model displays a striking resemblance to the fibrotic foci seen in the IPF fibrotic phenotype in the clinic - including apoptosis-resistant, hyperproliferative fibroblasts an exaggerated extracellular matrix accumulation. The assay will be validated with tool compounds and functional genomics probes available at the Molecular Screening Shared Resource at UCLA. 2. We will subject our disease model to a phenotype-based small molecule primary pilot HTS screen with high content screening (HCS) as readout. HCS is the ideal tool for assessing the activity of small molecules in this context as the disease modifying targets of IPF are unknown but the changes in cell morphology from the diseased to healthy state are very obvious. 3. We will perform hit validation, secondary assays/hit triaging and characterization through counterscreening, toxicity testing and functional genomics such as lentiviral shRNA gene knockdown and cDNA complementation strategies. We aim to identify novel small molecule probes, approved drugs and new small molecules that target the abnormal tissue remodeling of IPF, which is characterized by excessive extracellular matrix accumulation. Thus this proposal aims to advance the diagnosis, treatment and prevention of the disease of IPF and is therefore fully aligned with the goals of the NIGMS.

描述（由适用提供）：特发性肺纤维化（IPF）是一种未知病因的疤痕疾病，每年在美国杀死40,000多名患者。该疾病的特征是纤维化灶中的肌纤维细胞聚集，这种肺重塑导致由于缺乏治疗选择，气体交换不足，这是渐进性且总是致命的。由于缺乏良好的测定系统复制疾病以及缺乏良好的模型系统来研究药物效率，因此对IPF发病机理的理解非常有限。我们使用IPF患者的诱导多能干细胞（IPSC）为IPF生成了一种新颖的，可靠的临床相关的人类模型。具体而言，随着患者衍生的IPSC衍生成菜肴的纤维化灶，当将其重新分化为间充质型细胞时，该模型将自己置于药物发现，从而在菜肴中进行了促进性纤维化灶，从而在菜肴中概括了该疾病表型。我们的具体目的是：1。我们将开发高吞吐量筛选（HTS）测定法，以识别针对目标发现和验证的小分子问题，测试已经批准为重新利用IPF治疗的药物，并添加了利用我们的“疾病”模型的IPF药物开发的新化合物。我们的体外模型表现出与临床中IPF纤维化表型中看到的纤维化焦点的显着相似之处 - 包括抗凋亡的，超增强的成纤维细胞，形成了夸张的细胞外基质积累。该测定法将通过UCLA分子筛选共享资源可用的工具化合物和功能基因组学项目进行验证。 2。我们将使疾病模型接受基于表型的小分子初级试验HTS筛选，其中高含量筛选（HCS）作为读数。 HCS是在这种情况下评估小分子活性的理想工具，因为修改IPF的疾病靶标是未知的，但是从解散到健康状态的细胞形态变化非常明显。 3。我们将通过截面，毒性测试和功能基因组学（例如慢病毒SHRNA基因敲低和cDNA完成策略）进行命中验证，次要测定/命中分三位生和表征。我们旨在确定针对IPF异常组织重塑的新型小分子问题，认可的药物和新的小分子，其特征是细胞外基质积累过多。该建议旨在提高IPF疾病的诊断，治疗和预防，因此与Nigms的目标完全一致。