Lipidomic Screening For Functional Surfactant Gene Mutations

功能性表面活性剂基因突变的脂质组学筛选

基本信息

批准号：
9021312
负责人：
Francis Sessions Cole
金额：
$ 45.57万
依托单位：
WASHINGTON UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-03-01 至 2019-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9021312
关键词：
1 year old ABCA3 gene Adenovirus Vector Alveolar Animals Award Binding Biological Models Cause of Death Cell Culture System Cell Culture Techniques Cells Complementary DNA Computer Simulation Defect Development Diagnostic Electron Microscopy Electrospray Ionization Epithelial Epitopes Family Fetal Lung Frequencies Gene Mutation Gene Silencing Genes Genetic Genomics Goals Health Heritability Human Individual Infant Kinetics Label Link Lipids Mass Fragmentography Mass Spectrum Analysis Measurement Mediating Methods Modeling Morbidity - disease rate Mutation Phase Phenotype Phospholipids Premature Infant Property RNA Regulation Resistance Risk Sensitivity and Specificity Site-Directed Mutagenesis Surface Surface Tension System Testing Treatment Failure United States Variant abstracting alveolar lamellar body base burden of illness disorder risk endoplasmic reticulum stress fetal gender disparity high throughput screening improved innovation lipid transport member mutation screening neonatal respiratory distress precursor cell racial disparity reconstitution respiratory respiratory distress syndrome screening small hairpin RNA small molecule species difference surfactant surfactant deficiency surfactant function surfactant production surfactant replacement tandem mass spectrometry tissue culture trafficking transduction efficiency treatment strategy

项目摘要

DESCRIPTION (provided by applicant): Genetic regulation of surfactant deficiency has been suggested by heritability of neonatal respiratory distress syndrome, persistence of gender and racial disparities in disease risk, and the frequency of surfactant replacement treatment failures. Individually rare, collectively common (2-4/100 infants) mutations in the ATP- binding cassette sub-family A member 3 gene (ABCA3) disrupt surfactant function through diverse mechanisms including reduced lipid transport, ABCA3 misfolding or altered trafficking, or induction of increased endoplasmic reticulum stress and increase risk for neonatal respiratory distress syndrome in term and late preterm infants. Our goal is to develop and implement a human model system that uses static and dynamic lipidomic signatures for functional screening of ABCA3 mutations and that could be used for testing small molecules to correct mutation-encoded, functional defects in any gene expressed in the human alveolar type 2 cell. In the R21 Phase of this Award, using adenoviral vectors with high transduction efficiency and cargo capacity and highly sensitive and specific mass spectrometry based lipidomic profiling, we will test the hypothesis that ABCA3 gene silencing and rescue with wild-type ABCA3 cDNA reconstitute surfactant phospholipid signatures in human, primary alveolar type 2 cells. In the R33 Phase of this Award, using mass spectrometry-based lipidomic profiling, electron microscopy, and surface activity measurements, we will examine disruption of surfactant lipidomic signatures, lipid turnover rates, and lipid secretion kinetics, lamellar body phenotype, and surfactant function by previously characterized and uncharacterized ABCA3 mutations associated with increased risk for neonatal respiratory distress syndrome. The overall impact of this Award will provide a human model system for functional, lipidomics-based screening of genomic hits associated with surfactant deficiency and for development of small- molecule based strategies to correct mutation-encoded, functional surfactant defects.

描述（由申请人提供）：遗传活性剂缺乏症的遗传调节是由于新生儿呼吸窘迫综合征的遗传力，性别和种族差异的持续性以及疾病风险中的种族差异的持续性以及表面活性剂替代治疗失败的频率。 Individually rare, collectively common (2-4/100 infants) mutations in the ATP- binding cassette sub-family A member 3 gene (ABCA3) disrupt surfactant function through diverse mechanisms including reduced lipid transport, ABCA3 misfolding or altered trafficking, or induction of increased endoplasmic reticulum stress and increase risk for neonatal respiratory distress syndrome in term and late preterm infants.我们的目标是开发和实施一个人类模型系统，该系统使用静态和动态的脂肪组信号来筛选ABCA3突变的功能性筛选，并且可用于测试小分子以纠正在人肺泡2细胞中表达的任何基因中的突变编码的功能缺陷。 In the R21 Phase of this Award, using adenoviral vectors with high transduction efficiency and cargo capacity and highly sensitive and specific mass spectrometry based lipidomic profiling, we will test the hypothesis that ABCA3 gene silencing and rescue with wild-type ABCA3 cDNA reconstitute surfactant phospholipid signatures in human, primary alveolar type 2 cells.在该奖项的R33阶段中，使用基于质谱的脂肪组分析，电子显微镜和表面活性测量值，我们将检查表面活性剂脂质分组信号的破坏，脂质离职率，脂质分泌率以及脂质分泌动力学，通过以前的表型和表面型互为构成cotrication contricatife croutized cottric croutized croticatife croutized cotca croutized cotca croutized cottrication contricification，遇险综合征。该奖项的总体影响将为基于脂肪症的功能，基于脂肪委员会的筛查提供与表面活性剂缺乏相关的基因组打击以及基于小分子的策略的发展，以纠正突变编码的功能表面活性剂缺陷。