Study of iPS cells-derived neural and glial cells from subjects with Monge's disease

对来自蒙日氏病受试者的 iPS 细胞衍生的神经细胞和神经胶质细胞的研究

基本信息

批准号：
9314960
负责人：
Gabriel G Haddad
金额：
$ 23.25万
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：
2017
资助国家：
美国
起止时间：
2017-02-01 至 2019-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9314960
关键词：
Acid Fast Bacillae Staining Method Acids Address Affect Altitude Altitude Sickness Astrocytes Biopsy Blood - brain barrier anatomy Brain Brain Hypoxia-Ischemia Calcium Cell Death Cell membrane Cells Cerebral Edema Cessation of life Chronic Confusion Cytosol Data Disease Dizziness Endothelial Cells Equilibrium Exhibits Exploratory/Developmental Grant Extracellular Space Fibroblasts Functional disorder Future Genes Genetic Goals Headache Homeostasis Human Hypoxia Impairment In Vitro Knock-out Lead Magnetic Resonance Imaging Mitochondria Mitochondrial Matrix Modeling Monitor NHE1 Neuroglia Neurologic Symptoms Neurons Pathogenesis Pathology Patients Peruvian Pharmacology Phenotype Play Proteins RNA Interference Regulation Role Sea Skin Sleep disturbances Sodium Bicarbonate Sodium-Hydrogen Antiporter Stress Symptoms Testing Therapeutic Therapeutic Agents Water Western Blotting Work base brain cell calcium uniporter cell injury design experimental study imaging system induced pluripotent stem cell knock-down mRNA Expression nerve stem cell novel novel therapeutics protein expression symporter

项目摘要

ABSTRACT The overall goal of this proposal is to better understand how the brain is affected in Chronic Mountain Sickness (CMS) or Monge's disease. The brain is one of the major targets of CMS, as manifested by the frequently seen CNS symptoms such as headache, dizziness, sleep disturbance, and mental confusion. MRI studies have revealed cerebral edema and multiple ischemic foci in the brain of CMS patients, suggesting that brain cell damage/death occurs in CMS which may lead to the above neurological symptoms. Therefore, understanding the vulnerability of brain cells to hypoxia in CMS is critical for developing the therapeutic strategy of CMS. Recently, we obtained skin biopsies from CMS patients and non-CMS healthy highlanders and re-programmed the skin cells into induced pluripotent stem cells (iPSCs). The iPSCs were then differentiated into neuroprogenitor cells and further into astrocytes and neurons. We then examined the acid- base regulatory mechanisms and the tolerance/vulnerability to hypoxia/ischemia stress in these astrocytes/neurons. Our preliminary data show that astrocytes derived from CMS exhibited increased acid extrusion rate, and CMS astrocytes/neurons were more vulnerable to hypoxic/ischemic challenges than non- CMS cells. Therefore, we hypothesize that astrocytes/neurons in CMS patients have altered acid-base regulatory mechanisms which render neurons/astrocytes more vulnerable to hypoxia. We will test this hypothesis via two specific aims: 1) examine the expression and activities of NHE1 and NBCe1 in CMS and non-CMS astrocytes/neurons under normoxia and hypoxia conditions; 2) investigate the role of NHE1 and NBCe1 in the cell death of CMS astrocytes/neurons following hypoxia. We will use H+ and Na+ imaging system together with the genetic knockdown and pharmacological inhibition of NHE1 and NBCe1 to determine their role in the tolerance/vulnerability of CMS astrocytes/neurons to hypoxic stress. A novel feature of our study is the establishment of an in vitro CMS model that was directly derived from CMS and non-CMS highlanders. It is highly likely from our studies that we will make a number of discoveries that will help us design better CMS therapeutic strategies.

抽象的该提案的总体目标是更好地了解慢性山中大脑如何受到影响疾病 (CMS) 或蒙日氏病。大脑是 CMS 的主要目标之一，这表现在常见的中枢神经系统症状，如头痛、头晕、睡眠障碍和精神错乱。核磁共振成像研究发现 CMS 患者脑部出现脑水肿和多处缺血灶，表明 CMS发生脑细胞损伤/死亡，可能导致上述神经系统症状。所以，了解 CMS 中脑细胞对缺氧的脆弱性对于开发治疗方法至关重要 CMS 策略。最近，我们获得了 CMS 患者和非 CMS 健康高地人的皮肤活检并将皮肤细胞重新编程为诱导多能干细胞（iPSC）。当时的 iPSC 分化为神经祖细胞，并进一步分化为星形胶质细胞和神经元。然后我们检查了酸- 这些基础调节机制和对缺氧/缺血应激的耐受性/脆弱性星形胶质细胞/神经元。我们的初步数据表明，源自 CMS 的星形胶质细胞表现出酸度增加挤出率，CMS 星形胶质细胞/神经元比非星形胶质细胞更容易受到缺氧/缺血的挑战。 CMS 细胞。因此，我们假设 CMS 患者的星形胶质细胞/神经元发生了酸碱度改变。使神经元/星形胶质细胞更容易缺氧的调节机制。我们将测试这个通过两个具体目标提出假设：1）检查 CMS 中 NHE1 和 NBCe1 的表达和活性，常氧和缺氧条件下的非 CMS 星形胶质细胞/神经元； 2）研究NHE1的作用和 NBCe1 在缺氧后 CMS 星形胶质细胞/神经元细胞死亡中的作用。我们将使用 H+ 和 Na+ 成像系统结合 NHE1 和 NBCe1 的基因敲除和药理学抑制，以确定它们的 CMS 星形胶质细胞/神经元对缺氧应激的耐受性/脆弱性中的作用。我们研究的一个新特点是建立直接源自 CMS 和非 CMS 高地犬的体外 CMS 模型。这是从我们的研究中，我们很可能会取得许多发现，这将有助于我们设计更好的 CMS 治疗策略。