Genetic Mechanisms Regulating Hypoxia Tolerance in the Brain

调节大脑缺氧耐受性的遗传机制

基本信息

批准号：
9894142
负责人：
Gabriel G Haddad
金额：
$ 43.36万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN DIEGO
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-04-01 至 2022-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9894142
关键词：
Abbreviations Acute Altitude Andean Animals Blood Blood Vessels Brain Brain Hypoxia-Ischemia CRISPR/Cas technology Candidate Disease Gene Cell Hypoxia Cell physiology Cells Cerebral Ischemia Chronic Clinical Comparative Genomic Analysis Development Directed Molecular Evolution Disease Drosophila genus Drosophila melanogaster Elements Ethiopian Evolution Excision GATA3 gene Gene Expression Generations Genes Genetic Genome Genomic approach Goals Healthcare Homo sapiens Human Human Genome Hypoxia Individual Injury Intervention Ischemic Stroke Laboratories Lead Lung diseases Malignant Neoplasms Medical Modeling Morbidity - disease rate Myocardial Infarction Neuroglia Neurons Organism Outcome Oxygen Pathogenesis Pathologic Predisposition Prevention Protein Isoforms Research Role Solid Neoplasm Stroke System Therapeutic Thinness Tissues Translating Treatment Efficacy Whole Organism Work clinical translation comparative genomics design experimental study fly genome analysis high reward high risk human disease improved in vivo induced pluripotent stem cell knock-down mortality novel overexpression pressure stroke therapy whole genome

项目摘要

Hypoxia represents a critical element in the pathogenesis of many human diseases, such as ischemic stroke, myocardial infarction and cancer and solid tumors. Understanding the mechanisms regulating hypoxia tolerance or susceptibility is essential for developing effective strategies for medical interventions. In this regard, we have generated a hypoxia-tolerant Drosophila melanogaster strain through laboratory-directed evolution (over >300 generations in descending environmental O2 levels with every several generations), sequenced their genomes and analyzed them in these flies. In parallel, we took advantage of the natural experimental evolution of humans in high-altitude regions of the world by sequencing and analyzing the whole genomes of Ethiopian and Andean highlanders. Through a comparative genomic approach combining our results with those of others, we obtained a group of evolutionarily conserved genes (28 human/23 Drosophila genes) that are potentially involved in regulating hypoxia tolerance in human highlanders and hypoxia-tolerant flies. Indeed, we discovered that ubiquitous knock-down of the expression of genes (Dm/Human), i.e., grn/GATA3, Mkk4/MAP2K4, pyd/TJP1, and shep/RBMS3, dramatically enhanced hypoxia tolerance in vivo in Drosophila. These mechanisms have a strong potential to be translated into novel targets for developing therapeutic strategies to treat hypoxia-related diseases. Our central hypothesis is that the group of evolutionary conserved genes regulates hypoxia tolerance in neurons and glial cells in humans. Our specific aims are: 1) To determine the role of evolutionarily conserved candidate genes (obtained from our Results in D. melanogaster and human highlanders) in hypoxia tolerance in vivo in whole organisms. We will determine the role of 23 candidate genes individually in hypoxia tolerance with ubiquitous or tissue/cell-specific knocking-down or overexpression using UAS/Gal4 system in D. melanogaster, and 2) To elucidate the role of evolutionarily conserved candidate genes in hypoxia tolerance in human iPSC-derived neuronal and glial cells. We will delineate the specific role of the candidate genes in protecting human neuronal and glial cells against hypoxia-induced injury by altering their expression using CRISPR/Cas9 system. We believe that this high-risk high-reward project will provide novel information about the mechanisms underlying hypoxia tolerance or vulnerability.

缺氧是许多人类疾病发病机制的关键因素，例如缺血性心脏病中风、心肌梗塞、癌症和实体瘤。了解调节缺氧的机制耐受性或敏感性对于制定有效的医疗干预策略至关重要。在这个对此，我们通过实验室指导培育出了耐缺氧的黑腹果蝇菌株进化（超过 300 代，环境 O2 水平每几代就会下降），对这些果蝇的基因组进行了测序并进行了分析。与此同时，我们利用自然的优势通过对整个世界高海拔地区的人类进行测序和分析，进行实验进化埃塞俄比亚和安第斯高地人的基因组。通过比较基因组方法结合我们的与其他人的结果相比较，我们获得了一组进化上保守的基因（28个人类/23个果蝇）基因）可能参与调节人类高地人和耐缺氧能力苍蝇。事实上，我们发现基因（Dm/人类）表达的普遍抑制，即 grn/GATA3、Mkk4/MAP2K4、pyd/TJP1 和 shep/RBMS3 显着增强了体内缺氧耐受性果蝇。这些机制有很大潜力转化为开发新目标治疗缺氧相关疾病的治疗策略。我们的中心假设是进化保守基因调节人类神经元和神经胶质细胞的缺氧耐受性。我们的具体目标是： 1）确定进化上保守的候选基因的作用（从我们在黑腹果蝇和人类高地人中的结果在体内耐缺氧方面的整体表现有机体。我们将分别确定 23 个候选基因在普遍存在的缺氧耐受性中的作用或使用 UAS/Gal4 系统在黑腹果蝇中进行组织/细胞特异性敲低或过度表达，以及 2) 阐明进化上保守的候选基因在人 iPSC 衍生的神经元和神经胶质细胞耐缺氧中的作用。我们将描述候选基因在保护中的具体作用使用 CRISPR/Cas9 改变人类神经元和神经胶质细胞的表达来抵抗缺氧引起的损伤系统。我们相信这个高风险高回报的项目将提供有关机制的新颖信息潜在的缺氧耐受性或脆弱性。