Genetic Mechanisms Regulating Hypoxia Tolerance in the Brain

缺氧是许多人类疾病发病机制的关键因素，例如缺血性疾病 中风、心肌梗塞和癌症以及实体瘤。了解调节缺氧的机制 耐受性或易感性对于制定有效的医疗干预策略至关重要。在这 关于这一点，我们已经通过实验室定向培养， 进化（超过300代，每隔几代环境O2水平下降）， 对它们的基因组进行了测序并分析了这些果蝇。与此同时，我们利用了 通过测序和分析整个人类在世界高海拔地区的实验进化 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 结果与其他研究结果比较，我们获得了一组进化上保守的基因（28个人/23个果蝇 基因），这些基因可能参与调节人类高原人的缺氧耐受性， 苍蝇事实上，我们发现基因表达的普遍敲低（Dm/人），即， grn/GATA 3、Mkk 4/MAP 2K 4、pyd/TJP 1和shep/RBMS 3显著增强了小鼠体内缺氧耐受性。 果蝇这些机制具有很强的潜力，可以转化为开发新的靶点。 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 具体目标是：1）确定进化上保守的候选基因（从 我们在D中的结果黑腹果蝇和高原人）在体内整体耐缺氧能力的研究 有机体我们将确定23个候选基因在缺氧耐受中的作用， 或利用UAS/Gal 4系统在D.黑腹，和2）To 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中的缺氧耐受性中的作用。我们将描述候选基因在保护这些基因中的具体作用。 人类神经元和神经胶质细胞通过使用CRISPR/Cas9改变其表达来对抗缺氧诱导的损伤 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信息， 潜在的缺氧耐受性或脆弱性。

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.