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) 是一种复杂且毁灭性的疾病，相关死亡率徘徊在 30-40%。即使补充氧气（此类患者的常规且必需的治疗方法）也会导致肺损伤。事实上，O2 的有害影响已将高氧性急性肺损伤 (HALI) 作为研究实验动物呼吸窘迫综合征的原型。为了应对高 ALI 死亡率并以不同于当前策略的方式评估 HALI 的遗传复杂性，我们建立了小鼠模型（敏感 C57BL/6J 和耐药 129X1/SvJ 小鼠），长期目标是识别基因和影响菌株存活差异的相关病理机制。对这些品系之间四种可能的杂交产生的 840 只 F2 小鼠进行的分离分析证实，存活时间是一个复杂的性状，外显率降低，并且具有显着的性别、杂交和亲本效应。对 840 只 F2 小鼠的数量性状基因座 (QTL) 分析确定了整个 F2 群体中的三个高度显着基因座（名为 Shali1-3，代表高氧急性肺损伤生存）和一个显着基因座 (Shali4)，以及一个显着的雄性特异性基因座基因座（Shali5）。配对分析确定了 QTL 之间的几种基因-基因相互作用以及与其他不连锁基因座的上位相互作用。分离和 QTL 分析表明，抗性等位基因均源自亲本菌株，并通过重组来确定个体的 HALI 易感性。这些结果得出以下假设：Shali QTL 含有易感基因，这些易感基因单独和/或以适当的等位基因组合组合在一起时，将显着影响 HALI 存活时间。该应用的主要目标是为物理作图和数量性状基因鉴定奠定基础，主要关注 Shali1。为了实现这一目标，我们提出了 3 个具体目标：1）通过为 B-S 模型中与 HALI 存活时间显着相关的 5 个 Shali QTL 构建互惠同源株来确认体内 QTL 结果；确定哪些 QTL 对 HALI 存活时间有显着贡献； 2) 测试候选基因，将Shali1 QTL区间缩小到适合物理作图的水平；优先考虑并严格评估候选基因的功能意义；同时，通过构建和检测同源亚系，缩短Shali1 QTL区间； 3) 确定在适当菌株中含有相应 QTL 的多同源菌株中提高和降低存活率的最佳等位基因组合；与同一菌株中所有四个敏感或所有四个抗性 Shali 等位基因产生相互同源。来自这些研究的小鼠品系将为我们提供所需的工具来识别和表征影响 HALI 存活的关键基因。公众健康启示：在我们建立的小鼠模型中使用 >95% 的氧气（高氧），该项目的长期目标是确定与急性肺损伤存活相关的关键基因，这将使我们能够将未来的努力集中在分子机制上涉及。我们之前的遗传分析发现了五个与生存时间显着相关的染色体区域；这些区域被命名为 Shali1-5，即高氧急性肺损伤存活基因座 1-5。该应用的主要目的是通过生成和测试固定背景中包含易感性等位基因的基因精炼小鼠模型，为识别控制生存的主要基因奠定基础。