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Pre-Clinical Models of VILI /ARDS Core

VILI /ARDS Core 的临床前模型

基本信息

批准号：
10094244
负责人：
Jason X J Yuan
金额：
$ 24.66万
依托单位：
UNIVERSITY OF ARIZONA
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-02-05 至 2021-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10094244
关键词：
Acute Lung Injury Adult Respiratory Distress Syndrome Agonist Animal Model Animals Arizona Basic Science Biochemical Biological Biological Assay Blood Vessels Breeding Caring Cell model Cells Cellular Assay Chemicals Chemistry Clinical Complementary DNA Complication Data Analyses Data Storage and Retrieval Development Endothelial Cells Endothelium Epigenetic Process Epithelial Equipment Escherichia coli Exposure to Functional disorder Genes Genetic Genetically Engineered Mouse Genotype Goals Histologic Housing Individual Infection Inflammation Intervention Knockout Mice Leadership Life Lipopolysaccharides Liposomes Lung Lung Inflammation Maintenance Measurement Mechanical Stress Mediating Microscopy Modeling Molecular Mus Mutate PBEF Gene Pathogenicity Patients Performance Pharmacology Phenotype Plasmids Post-Translational Protein Processing Pre-Clinical Model Proteins Protocols documentation Receptor Signaling Reproducibility Research Personnel Resources Role SOX18 gene Sampling Sepsis Signal Transduction Single Nucleotide Polymorphism Site Small Interfering RNA Sphingosine-1-Phosphate Receptor Syndrome Techniques Therapeutic Therapeutic Agents Therapeutic Effect Tissue Sample Tissues Training Transgenic Mice Translations Universities Vascular Endothelial Cell Vascular Permeabilities Ventilator Ventilator-induced lung injury Western Blotting Work animal breeding animal care antibody conjugate clinically relevant conditional knockout data sharing design efficacy testing experience experimental study high standard immunocytochemistry in vivo imaging insight lung injury lung preservation mouse model nitration novel novel therapeutic intervention pneumonia model pre-clinical preclinical efficacy prevent programs targeted agent technique development tool transcription factor vascular inflammation

项目摘要

SUMMARY: Core C, Pre-Clinical Models of ventilator-induced lung injury (VILI)/acute respiratory distress syndrome (ARDS), is designed to provide PPG investigators with rigorously defined and reproducible murine models of VILI, a two-hit lung injury model induced by exposure to both a ventilator and lipopolysaccharide (LPS) administration (VILI+LPS) that better mimics ARDS in patients, and an E. coli pneumonia model. Core C will comprehensively generate, manage and provide all animal-related experiments, resources, and expertise by accomplishing 6 specific aims. Specific Aim #1 will provide a complete range of expertise, training, equipment, and data analysis tools to extensively study the pathogenic mechanisms in preclinical models of murine lung injury. We will employ state-of-the-art techniques to a) characterize the role of various intracellular signaling cascades in regulating lung endothelial cell (EC) barrier function, b) determine the effects of specific interventions to provide insight into the efficacy and mechanisms of novel therapeutic strategies; and c) facilitate the translation of basic research to clinical interventions. Toward these goals, Core C will first provide validated quantitative measurements of vascular permeability and inflammation. Specific Aim #2 will house and care for the genetically-engineered mice and to generate novel transgenic and knockout mice (e.g., inducible endothelium-specific and lung epithelium-specific conditional knockout mice). Specific Aim #3 will examine selective siRNAs or pharmacological agents for target signaling cascades as potential therapeutic strategies and approaches for preclinical models and ultimately, for ARDS. Specific Aim #4 will be to provide performance of specific experimental strategies involving VILI, VILI+LPS (“two-hit”), E. coli pneumonia models of ARDS as well as gene-specific rescue interventions. Specific Aim #5 will evaluate the function of ARDS- associated single nucleotide polymorphisms (SNPs) and sites of functional protein post-translational modification (PTM), utilizing mutated cDNA (high efficiency expression plasmids) targeting the lung endothelium (with ACE antibody-conjugated liposome) in the endothelial conditional knockout mice. Specific Aim #6 will provide high quality shared data for data storage and well-preserved lung tissue samples to individual projects for more sophisticated molecular and cellular assays. Core C will centralize all mice-related work across all three projects of this program, including generating new strains, breeding and housing of mice, generating preclinical VILI/ARDS and VILI+LPS models, accessing therapeutic effects of siRNAs and chemicals, performing lung inflammation assessment, and providing tissue samples and freshly dissociated murine lung vascular endothelial cells to each project for specific assays (including immunohisto- and immunocytochemistry, western blot analysis, fluorescent microscopy). In addition to its own space and equipment, Core C will have full access to and will utilize resources available at the University of Arizona shared facilities including the Genetically Engineered Mouse Models Core (GEMM).

总结：核心C，呼吸机诱导的肺损伤（VILI）/急性呼吸窘迫综合征的临床前模型（ARDS），旨在为PPG研究者提供严格定义的和可重复的小鼠模型， VILI，由暴露于呼吸机和脂多糖（LPS）诱导的二次打击肺损伤模型给予（VILI + LPS）更好地模拟患者中的ARDS，以及E.大肠杆菌肺炎模型。核心C将全面生成，管理和提供所有与动物相关的实验，资源和专业知识，实现6个具体目标。具体目标#1将提供全面的专业知识，培训，设备和数据分析工具，以广泛研究临床前模型中的致病机制，小鼠肺损伤我们将采用最先进的技术来a）表征各种细胞内的作用，信号级联在调节肺内皮细胞（EC）屏障功能中的作用，B）确定特异性干预措施，以深入了解新的治疗策略的疗效和机制;以及c）促进基础研究转化为临床干预措施。为了实现这些目标，Core C将首先提供经验证的血管渗透性和炎症的定量测量。具体目标#2将容纳并照顾基因工程小鼠和产生新的转基因和敲除小鼠（例如，可诱导内皮特异性和肺上皮特异性条件性敲除小鼠）。具体目标#3 检查选择性siRNA或药理学试剂作为潜在的治疗靶向信号级联反应临床前模型的策略和方法，并最终用于ARDS。具体目标#4将提供包括VILI、VILI + LPS（"两次打击"）、E.大肠杆菌肺炎模型以及基因特异性抢救干预。具体目标#5将评估ARDS的功能- 相关的单核苷酸多态性（SNP）和功能蛋白翻译后位点修饰（PTM），利用靶向肺的突变cDNA（高效表达质粒在内皮条件性敲除小鼠中的内皮（用ACE抗体缀合的脂质体）。具体目标6将为数据存储和保存良好的肺组织样本提供高质量的共享数据，用于更复杂的分子和细胞分析的单独项目。核心C将集中所有与老鼠有关的在该计划的所有三个项目中开展工作，包括产生新的品系，饲养和饲养小鼠，产生临床前VILI/ARDS和VILI + LPS模型，评估siRNA的治疗效果，化学品，进行肺部炎症评估，并提供组织样本和新鲜分离的小鼠肺血管内皮细胞，以每个项目的特定测定（包括免疫组化和免疫细胞化学、蛋白质印迹分析、荧光显微术）。除了自己的空间和设备，核心C将有充分的机会，并将利用亚利桑那大学的资源共享设施，包括基因工程小鼠模型核心（GEMM）。