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Mechanistic roles of Cytochrome P4501A enzymes in hyperoxic lung injury

细胞色素 P4501A 酶在高氧性肺损伤中的机制作用

基本信息

批准号：
9127549
负责人：
BHAGAVATULA MOORTHY
金额：
$ 54.03万
依托单位：
BAYLOR COLLEGE OF MEDICINE
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-04-01 至 2020-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9127549
关键词：
8-hydroxyguanosine Acute Lung Injury Adult Adult Respiratory Distress Syndrome Air Albumins Antibodies Aspirate substance Binding Biological Assay Biological Markers Bronchopulmonary Dysplasia CYP1A1 gene CYP1A2 gene Carbazoles Cells Cytochrome P450 Cytochromes DNA Adducts DNA Damage DNA lesion Development Dysplasia Enzymes Epoxy Compounds Genes Genetic Transcription Genomic DNA Glycols Hepatic Human Hyperoxia Immunohistochemistry Immunoprecipitation In Vitro Infant Injury Knock-in Mouse Knockout Mice Laboratories Lead Ligands Linoleic Acids Lipids Liver Lung Measures Mediating Modeling Molecular Mus Oxygen Patients Phase Predisposition Premature Infant Prevention Prevention strategy Protein Array Proteomics Pulmonary Valve Insufficiency Reactive Oxygen Species Research Role System Techniques Testing Time Tissues Transgenic Mice Wild Type Mouse adduct base endotracheal in vitro Bioassay in vivo indexing leukotoxin lung injury metabolomics mouse model novel novel marker novel strategies oxidative DNA damage promoter public health relevance receptor receptor binding translational study

项目摘要

DESCRIPTION (provided by applicant): Hyperoxia is frequently used in the treatment of pulmonary insufficiency in premature infants and adults with acute lung injury (ALI) and/or acute respiratory distress syndrome (ARDS). However, in infants, hyperoxia contributes to the development of Broncho pulmonary dysplasia (BPD) in infants, and exacerbates lung injury in ALI/ARDS patients. Studies from our laboratory have clearly shown a mechanistic role for cytochrome P450 (CYP)1A enzymes in oxygen injury. The central hypothesis of this research is that hyperoxia induces pulmonary cytochrome P450 (CYP)1A enzymes in vivo by forming novel endogenous Ah receptor (AHR) ligands, and that hepatic CYP1A2 protects against hyperoxic lung injury by metabolizing reactive oxygen species (ROS)-mediated molecules and/or linoleic acid-derived leukotoxin epoxides or diols) that cause lung injury. The hypothesis that specific ROS-mediated oxidative DNA lesions in humans could serve as novel biomarkers of human ALI/ARDS will also be tested. In order to test the above-mentioned hypotheses, we propose the following Specific Aims: 1. To test the hypothesis that 6-formyllindolo[3,2-b]carbazole (FICZ) is the novel endogenous ligand of the Ah receptor (AHR) that induces hepatic and pulmonary CYP1A enzymes in vivo and in vitro under hyperoxic conditions. This aim has two sub-aims: To elucidate the role of FICZ and/or AHR in the induction of human CYP1A by hyperoxia in vivo. (ii). To unravel the molecular mechanisms of CYP1A1 induction in human lung cells by hyperoxia, and test the hypothesis that FICZ (formed under hyperoxic conditions) contributes mechanistically to CYP1A1 induction. 2. To determine the molecular mechanisms by which liver CYP1A enzymes contribute to hyperoxic lung injury. We hypothesize that leukotoxin epoxides and diols will accumulate in lungs of Cyp1a2-null mice, and will contribute to the increased susceptibility to hyperoxic lung injury. We will create a humanized knock-in mouse model, which will express human CYP1A2 in a liver-specific manner (using Crealb transgenic mice driven by albumin promoter) in mice on a Cyp1a2-null background. We will determine if these mice will be rescued against hyperoxic lung injury. 3. To determine the mechanistic role of CYP1A enzymes in oxidative DNA damage-mediated by hyperoxia. This aim has two sub-aims: (i) to test the hypothesis that mice lacking the genes for CYP1A1 and or 1A2 are more susceptible to pulmonary oxidative DNA damage upon hyperoxic exposures than WT mice, and that novel oxidative DNA lesion mechanistically contribute to hyperoxic lung injury in mice and ARDS in humans. (ii) To test the hypothesis that endotracheal aspirates of ALI/ARDS patients will display specific oxidative DNA lesions that could serve as novel biomarkers for ALI/ARDS. Successful accomplishment of the aims could lead to the development of novel biomarkers of ALI/ARDS, and new strategies for the prevention and/or treatment of ALI/ARDS in humans.

描述（由申请方提供）：高氧常用于治疗患有急性肺损伤（ALI）和/或急性呼吸窘迫综合征（ARDS）的早产儿和成人的肺功能不全。然而，在婴儿中，高氧有助于婴儿支气管肺发育不良（BPD）的发展，并加重ALI/ARDS患者的肺损伤。我们实验室的研究已经清楚地表明了细胞色素P450（P450）1A酶在氧损伤中的机制作用。本研究的中心假设是，高氧通过形成新的内源性Ah受体（AHR）配体在体内诱导肺细胞色素P450（CYP）1A酶，肝CYP 1A 2通过代谢导致肺损伤的活性氧（ROS）介导的分子和/或亚油酸衍生的白细胞毒素环氧化物或二醇来保护高氧肺损伤。还将检验人类中特定ROS介导的氧化DNA损伤可作为人类ALI/ARDS的新生物标志物的假设。为了验证上述假设，我们提出了以下具体目标：1。为了验证假设6-甲酰吲哚并[3，2-B]咔唑（FICZ）是Ah受体（AHR）的新型内源性配体，在体内和体外高氧条件下诱导肝和肺CYP 1A酶。该目的有两个子目的：阐明FICZ和/或AHR在体内高氧诱导人CYP 1A中的作用。（二）.阐明高氧诱导人肺细胞CYP 1A 1的分子机制，并检验FICZ（在高氧条件下形成）在机制上促进CYP 1A 1诱导的假设。2.确定肝脏CYP 1A酶参与高氧肺损伤的分子机制。我们假设白细胞毒素环氧化物和二醇会在Cyp 1a 2基因缺失小鼠的肺中积累，并会导致对高氧肺损伤的易感性增加。我们将创建人源化敲入小鼠模型，该模型将在Cyp 1a 2缺失背景的小鼠中以肝脏特异性方式（使用白蛋白启动子驱动的Crealb转基因小鼠）表达人CYP 1A 2。我们将确定这些小鼠是否会被拯救以对抗高氧肺损伤。3.确定CYP 1A酶在高氧介导的DNA氧化损伤中的作用机制。这一目标有两个次级目标：（i）检验缺乏CYP 1A 1和/或1A 2基因的小鼠在高氧暴露时比WT小鼠更易受肺氧化DNA损伤的假设，以及新的氧化DNA损伤在机制上导致小鼠的高氧肺损伤和人类的ARDS。(ii)为了验证ALI/ARDS患者的气管内抽吸物将显示特异性氧化DNA损伤的假设，该损伤可作为ALI/ARDS的新生物标志物。这些目标的成功实现可能导致开发新的ALI/ARDS生物标志物，以及预防和/或治疗人类ALI/ARDS的新策略。