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的大脑如何影响大脑疾病（CMS）或Monge病。大脑是CM的主要目标之一，如经常看到CNS症状，例如头痛，头晕，睡眠障碍和精神混乱。 MRI 研究表明，CMS患者大脑中的大脑水肿和多重缺血灶，表明脑细胞损伤/死亡发生在CMS中，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发展治疗至关重要 CMS的策略。最近，我们从CMS患者和非CMS健康高地的皮肤活检中获得了皮肤活检并将皮肤细胞重新编程为诱导的多能干细胞（IPSC）。 IPSC是分化为神经元基因细胞，进一步为星形胶质细胞和神经元。然后我们检查了酸 - 基础调节机制以及对低氧/缺血压力的耐受性/脆弱性星形胶质细胞/神经元。我们的初步数据表明，源自CM的星形胶质细胞表现出酸的增加挤出率和CMS星形胶质细胞/神经元比非非 - CMS细胞。因此，我们假设CMS患者的星形胶质细胞/神经元改变了酸碱使神经元/星形胶质细胞更容易受到缺氧的调节机制。我们将测试这个通过两个具体目的假设：1）检查CMS和NBCE1和NBCE1的表达和活动在正常氧和缺氧条件下，非CMS星形胶质细胞/神经元； 2）研究NHE1和缺氧后CMS星形胶质细胞/神经元的细胞死亡中NBCE1。我们将使用H+和Na+成像系统加上NHE1和NBCE1的遗传敲低和药理抑制 CMS星形胶质细胞/神经元对低氧应激的耐受性/脆弱性中的作用。我们研究的新功能是建立了直接来自CMS和非CMS高地人的体外CMS模型。这是从我们的研究中，我们将有很多发现可以帮助我们设计更好的CMS的发现治疗策略。