Genetic Analysis of Hyperoxia Induced Acute Lung Injury

高氧所致急性肺损伤的基因分析

基本信息

批准号：
8251198
负责人：
Daniel R Prows
金额：
$ 38.63万
依托单位：
CINCINNATI CHILDRENS HOSP MED CTR
依托单位国家：
美国
项目类别：
财政年份：
2003
资助国家：
美国
起止时间：
2003-12-15 至 2013-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8251198
关键词：
129X1/SvJ Mouse Acute Lung Injury Address Adult Adult Respiratory Distress Syndrome Aerosols Affect Alleles Animals Backcrossings Candidate Disease Gene Cessation of life Clinical Research Complex Computer Simulation Congenic Strain Critical Illness Epigenetic Process Funding Future Genes Genetic Genetic Polymorphism Genetic Variation Genome Goals Health Human Hyperoxia Individual Inheritance Patterns Knowledge Link Modeling Molecular Monitor Mus Myocardial Infarction Names Nickel Other Genetics Outcome Oxygen Ozone Parents Pathologic Patients Penetrance Population Predisposition Premature Infant Quantitative Trait Loci Recombinants Research Resistance Resources Respiratory distress Role Sample Size Severities Staging Susceptibility Gene Testing Time congenic gene interaction genetic analysis improved in vivo lung injury male mortality mouse model novel physical mapping prototype resistance allele respiratory distress syndrome segregation sex tool trait

项目摘要

DESCRIPTION (provided by applicant): Acute lung injury (ALI) and its most severe presentation acute respiratory distress syndrome (ARDS), represent a full spectrum of a complex and devastating illness, with associated mortality hovering at 30-40%. Even supplemental O2, a routine and needed therapy for such patients, paradoxically causes lung injury. In fact, detrimental effects of O2 have established hyperoxic acute lung injury (HALI) as a prototype to study respiratory distress syndromes in experimental animals. To confront the high ALI mortality rate and to assess the genetic complexity of HALI differently than current strategies, we have established a mouse model (sensitive C57BL/6J and resistant 129X1/SvJ mice), with a long-term goal is to identify genes and the related pathologic mechanisms affecting strain survival differences. Segregation analysis of 840 F2 mice generated from the four possible intercrosses between these strains verified that survival time is a complex trait with reduced penetrance, and significant sex, cross, and parent-of-origin effects. Quantitative trait locus (QTL) analyses of the 840 F2 mice identified three highly significant loci (named Shali1-3, for Survival to hyperoxic acute lung injury) and one significant locus (Shali4) in the total F2 population, and a significant male-specific locus (Shali5). Pairwise analysis identified several gene-gene interactions among the QTLs and an epistatic interaction with an otherwise unlinked locus. Segregation and QTL analyses revealed that resistance alleles originate from both parental strains and recombine to determine individual HALI susceptibility. These results have led to the following hypothesis: Shali QTLs contain susceptibility genes that, separately and/or when grouped together in appropriate allelic combinations, will significantly affect HALI survival time. The primary objective of this application is to set the stage for physical mapping and quantitative trait gene identification, with a major focus on Shali1. To accomplish this, we propose 3 Specific Aims: 1) confirm QTL results in vivo by constructing reciprocal congenic strains for the five Shali QTLs significantly linked to HALI survival time in the B-S model; establish which QTL(s) significantly contribute to HALI survival time; 2) test candidate genes and reduce the Shali1 QTL interval to a level amenable to physical mapping; prioritize and critically assess candidate genes for functional significance; concurrently, reduce the Shali1 QTL interval by constructing and testing congenic substrains; and 3) determine the best allelic combinations for increased and decreased survival in multi-congenic strains containing the corresponding QTLs in the appropriate strain; generate reciprocal congenics with all four sensitive or all four resistant Shali alleles in the same strain. Mouse lines derived from these studies will give us the needed tools to identify and characterize the key gene(s) affecting HALI survival. PUBLIC HEALTH REVELANCE: Using >95% oxygen (hyperoxia) in our established mouse model, the long-term goal of this project is to identify critical genes involved in acute lung injury survival, which will allow us to focus future efforts on the molecular mechanisms involved. Our previous genetic analyses identified five chromosomal regions significantly linked with survival time; these regions were named Shali1-5, for Survival to hyperoxic acute lung injury loci 1-5. The primary objective of this application is to set the stage for identifying the major gene(s) controlling survival by generating and testing genetically-refined mouse models containing susceptibility alleles in a fixed background.

描述（由申请人提供）：急性肺损伤（ALI）及其最严重的急性呼吸窘迫综合征（ARDS），代表着复杂且毁灭性疾病的完整范围，相关的死亡率徘徊在30-40％。即使是补充O2，这是对此类患者的常规和必要治疗，也会矛盾地造成肺损伤。实际上，O2的有害作用已确立了高氧化急性肺损伤（HALI）作为研究实验动物中呼吸窘迫综合征的原型。为了面对高ALI死亡率并以与当前策略不同的是，对哈利的遗传复杂性不同，我们建立了小鼠模型（敏感的C57BL/6J和耐药129x1/SVJ小鼠），其长期目标是识别基因以及影响应变生存差异的相关病理机制。从这些菌株之间的四个可能的间交叉产生的840只F2小鼠的分离分析证实，生存时间是一个复杂的特征，具有降低的渗透率，显着的性别，交叉和父母 - 卵巢效应。对840 F2小鼠的定量性状基因座（QTL）分析鉴定出三个高度显着的基因座（称为Shali1-3，以生存到高氧化急性急性肺损伤）和一个在F2总体中的一个显着的基因座（SHALI4），一个显着的男性特异性基因座（SHALI5）。成对分析确定了QTL之间的几种基因 - 基因相互作用，以及与原本未链接基因座的上皮相互作用。隔离和QTL分析表明，抗性等位基因既起源于亲本菌株和重组，以确定单个哈利的敏感性。这些结果导致了以下假设：Shali QTL包含易感基因，这些基因分别和/或将其分组为适当的等位基因组合时，将显着影响Hali的生存时间。该应用的主要目的是为物理映射和定量性状基因识别设定阶段，重点是Shali1。为此，我们提出了3个具体目的：1）通过为五个shali QTL构建互惠的先天性菌株确认QTL导致体内产生QTL，这与B-S模型中的Hali生存时间显着相关；确定哪些QTL显着促进了哈利生存时间； 2）测试候选基因并将SHALI1 QTL间隔降低到适合物理映射的水平；优先和批判性地评估候选基因的功能意义；同时，通过构建和测试先天基质来减少SHALI1 QTL间隔； 3）确定最佳的等位基因组合，以增加和降低适当菌株中包含相应QTL的多综合菌株的存活率和降低；在同一菌株中产生具有所有四个敏感或所有抗性的Shali等位基因的互惠式友善。从这些研究中得出的小鼠线将为我们提供所需的工具，以识别和表征影响哈利生存的关键基因。公共卫生启示：在我们既定的小鼠模型中使用> 95％的氧（高氧），该项目的长期目标是确定涉及急性肺损伤生存的关键基因，这将使我们能够将未来的努力集中在涉及的分子机制上。我们以前的遗传分析确定了五个与生存时间显着相关的染色体区域。这些区域命名为Shali1-5，以生存到高氧性急性肺损伤基因座1-5。该应用程序的主要目的是为确定控制生存的主要基因的阶段，通过生成和测试含有固定背景下易感性等位基因的遗传精制的小鼠模型。