Functional Genomics of Chemical-Induced Acute Lung Injury

描述（由申请人提供）：化学诱导的急性肺损伤（CIALI）可能是由许多化学威胁引起的，这些化学威胁可用于恐怖袭击。需要治疗几种化学威胁剂共同的急性效应和病理。尽管付出了巨大的努力，但关于吸入暴露与迟发性肺水肿、呼吸衰竭和最终死亡之间的病理事件，仍有许多有待了解的地方。在过去的资助期内，我们开发并验证了5种常见化学物质（丙烯醛、氨、氯、光气和硫酸）急性肺损伤小鼠模型。使用功能基因组学方法，使用40个小鼠品系来鉴定与暴露后存活时间相关的候选基因。我们结合这些结果构建了一个蛋白质相互作用网络（interactome）。在这个网络中，发现了一个与v- AKT1胸腺瘤病毒致癌基因同源物1 （AKT1）相关的细胞信号中枢（即具有几种蛋白质相互作用的蛋白质）。当被磷酸化时，AKT1通过使细胞死亡机制的成分失活来增强存活。虽然在癌症中是不可取的，但我们认为短期、可逆的增强细胞存活AKT1活性可能对CIALI治疗有益。我们随后发现，抑制10号染色体上的磷酸酶和紧张素同源物缺失（PTEN），一种限制AKT1活性的磷酸酶，在体外增强上皮修复，并在体内预防CIALI。假设：抑制PTEN活性将通过激活促进细胞存活的信号事件赋予对CIALI的抗性。方法：采用高含量筛选方法，体外给药先导化合物（包括PTEN抑制剂），以测试靶细胞的致死性逆转。铅化合物将在小鼠模型中与5种化学物质一起测试CIALI的致死率。总目的：研究一种能提高多种化学物质致肺损伤患者生存率的治疗方法。

DESCRIPTION (provided by applicant): Chemical-induced acute lung injury (CIALI) can result from numerous chemical threats that avail themselves to terrorist attacks. Therapies are needed to treat the acute effects and pathologies that are common to several chemical threat agents. Despite intensive effort, much remains to be understood regarding pathological events linking Inhalation exposures to delayed pulmonary edema, respiratory failure, and ultimately death. In past funding period we developed and validated mouse models of acute lung injury to 5 common chemicals (acrolein, ammonia, chlorine, phosgene, and sulfuric acid). Using a functional genomics approach, 40 mouse strains were used to identify candidate genes associated with survival time following exposure. We combined the results to build a protein interaction network (interactome). Within this network, a cell signaling hub (i.e. a protein with several protein-protein interactions) was uncovered that implicated v- AKT1 thymoma viral oncogene homolog 1 (AKT1). When phosphorylated, AKT1 enhances survival by inactivating components of the cell death machinery. Although undesirable in cancer, we reasoned that short-term, reversible enhancement of the cell survival AKT1 activity could be beneficial in CIALI treatment. We subsequently found that inhibition of phosphatase and tensin homolog deleted on chromosome 10 (PTEN), a phosphatase that limits AKT1 activity, enhances epithelial repair in vitro and protect against CIALI in vivo. Hypothesis: Inhibition of PTEN activity will impart resistance to CIALI by activating signaling events that promote cell survival. Approach: Using a high content screening method, lead compounds [including a PTEN inhibitor] will be administered in vitro to test reverse of lethality in target cells. Lead compounds will be tested in mouse models of lethality from CIALI with 5 chemicals. Overall Objective: To develop a therapy that improves survival during lung injury induced by multiple chemicals. Public Health Relevance: Acute lung injury can result from numerous chemical threats that avail themselves to terrorist attacks. Current therapy remains limited to supportive care with no approved therapeutic for post insult treatment. The overall objective is to develop a therapy that will improve survival during acute lung injury induced by more than one chemical.