MENTORED PATIENT-ORIENTED RESEARCH CAREEER DEVELOPMENT AWARD

以患者为导向的研究职业发展奖

基本信息

批准号：
7231418
负责人：
Robert J Freishtat
金额：
$ 12.47万
依托单位：
CHILDREN'S RESEARCH INSTITUTE
依托单位国家：
美国
项目类别：
财政年份：
2005
资助国家：
美国
起止时间：
2005-09-23 至 2010-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7231418
关键词：
Accounting Acute Lung Injury Address Admission activity Adult Adult Respiratory Distress Syndrome Alveolar Alveolar wall Anatomy Area Award Biological Markers Blood Blood Cells Blood Platelets Blood specimen CD4 Positive T Lymphocytes CD40 Ligand CD8-Positive T-Lymphocytes Cell Communication Cell Nucleus Cells Child Childhood Chronic Disease Chronic Obstructive Airway Disease Clinical Data Clinical Research Coagulation Process Code Collection Critical Care Data Data Collection Data Set Development Diffuse Disease Disease Progression Disruption Endopeptidases Endothelium Enzyme-Linked Immunosorbent Assay Etiology Evaluation Evolution Fibrosis Flow Cytometry Fluorescence Foundations Functional disorder Future Gene Expression Gene Proteins Genes Genetic Genetic Polymorphism Genetic Transcription Genome Goals Heart Histocompatibility Antigens Class I Histologic Hour Human Human Genome Hypoxemia ICD-9 IL6 gene Immunity Individual Infiltration Inflammatory Inflammatory Response Institutes Intensive Care Units Intervention Investigation Knowledge Lead Light Liquid substance Literature Localized Longitudinal Studies Lung Lung diseases Lymphocyte Maps Mechanical ventilation Mediating Mediator of activation protein Membrane Mentors Modeling Molecular Molecular Profiling Mononuclear Morbidity - disease rate Natural Immunity Organ Outcome Pathogenesis Pathway interactions Patients Pattern Pediatric Intensive Care Units Peptide Hydrolases Perception Peripheral Phenotype Play Pneumonia Polyribosomes Principal Investigator Process Promoter Regions Proteins Pulmonary Edema Pulmonary Surfactant-Associated Protein B RNA RNA Stability Randomized Clinical Trials Rate Research Research Design Respiratory Failure Reverse Transcriptase Polymerase Chain Reaction Risk Role Sampling Score Sepsis Series Severities Severity of illness Single Nucleotide Polymorphism Staging Stress Supportive care Survivors System T-Lymphocyte Techniques Text Therapeutic Thrombosis Time Tobacco smoking Trauma Western Blotting Whole Blood Work base cell type cohort design experience human subject improved insight monocyte mortality neutrophil outcome forecast patient oriented research peripheral blood programs prospective protein expression research study tertiary care

项目摘要

Based on new insights into the molecular processes of AL! we hypothesize that a longitudinal analysis of cells of innate immunity and thrombosis will demonstrate changes in RNA and protein expression that are significantly associated with progression of All from sepsis. Hypothesis 1:The progression of sepsis- induced ALI is significantly associated with critical interactions between Th1 and Th2 lymphocytes and platelets. Specific Aim 1. We will collect total RNA from peripheral blood neutrophils, monocytes, TH1 and TH2 and CD8+ T lymphocytes and platelets from 20 mechanically ventilated pediatric patients with early ALI from sepsis. Blood sampling will take place at 6 individual time points over 72 hours along with severity of illness scoring to determine degree of disease progression. The temporal series of all 6 cell types banked for 3 retrospectively identified patients exemplary of ALI progressionwill be expression profiled and compared to profiles in 3 non-progressive ALI patients and 3 normal controls in order to generate a map of potential cell-cell interactions. Hypothesis 2: The pattern of peripheral blood cell expression of genes and proteins functionally important to the progression of ALI from sepsis will reflect the pattern of gene and protein expression in pulmonary edema cells and fluid. Specific Aim 2. Using the prioritization model from Aim 1,we will select the 2 cell types that show promise as "indicator" cell types. From the prospective cell collections in Aim 1 and more focused collections in this aim, we will choose, at random, 15 progressive and 15 non-progressive ALI subjects as determined by criteria outlined in the text of this proposal. We will verify select indicator gene expression changes found in Aim 1 by QMF RT-PCR in all 30 subjects (e.g. in the 20 genes with the most significant fold changes or most functional relevance.) Functionally important changes in gene expression will be confirmed in the entire cohort on the protein level by flow cytometry, and/or ELISA of proteins in peripheral blood and pulmonary edema fluid.

基于对Al分子过程的新见解！我们假设对先天免疫和血栓形成细胞将显示RNA和蛋白质表达的变化与败血症的所有进展显着相关。假设1：败血症的进展诱导的ALI与Th1和Th2淋巴细胞之间的关键相互作用显着相关血小板。具体目标1。我们将从外周血中性粒细胞，单核细胞，TH1和来自20名机械通风的小儿患者的Th2和CD8+ T淋巴细胞和血小板来自败血症的阿里。血液采样将在72小时内的6个单个时间点以及严重性进行疾病评分以确定疾病进展程度。所有6种细胞类型的时间系列为3个回顾性确定的Ali进展的患者示范，将表达表达，并与3名非促进ALI患者和3种正常对照的轮廓相比，为了生成一张地图潜在的细胞 - 细胞相互作用。假设2：基因外周血细胞表达的模式和蛋白质在功能上对ALI从败血症进展的功能很重要，将反映基因的模式和肺水肿细胞和液体中的蛋白质表达。特定目标2。使用来自的优先级模型 AIM 1，我们将选择2种显示有望为“指示”单元格类型的单元格。从前瞻性细胞 AIM 1中的集合和更集中的集合中的收集，我们将随机选择15个进步和15个非促进性ALI受试者，由本提案文本中概述的标准确定。我们将验证所有30个受试者在QMF RT-PCR中发现AIM 1中的某些指标基因表达变化（例如，在具有最显着倍数变化或功能最相关的20个基因中。）基因表达的重要变化将在整个队列中通过流量确认外周血和肺水肿流体中蛋白质的细胞仪和/或ELISA。