Molecular characterization of reactive astrocytes in humans

项目概要/摘要 星形胶质细胞构成人脑中所有细胞的至少三分之一，并且对于发育和生长至关重要。 中枢神经系统的功能。反应性星形胶质细胞增生是一系列细胞、分子和功能性 星形胶质细胞的变化在广泛的损伤和疾病中发现，包括癫痫，脑肿瘤， 阿尔茨海默氏病、帕金森氏病、中风、炎症和创伤性脑损伤。基于研究 在小鼠模型中，反应性星形胶质细胞在疾病进展和神经系统疾病中既有有益的作用，也有有害的作用。 通过分泌调节免疫细胞的细胞因子，产生生长因子和形成疤痕， 将疾病组织与健康组织隔离。然而，人们对分子和细胞的变化知之甚少， 在人类患者中的星形胶质细胞中，部分由于纯化和培养人类星形胶质细胞的困难。 先前纯化人星形胶质细胞的方法依赖于血清，其在星形胶质细胞中诱导反应性星形胶质细胞增生。 纯化程序，使得难以研究患者中星形胶质细胞的反应性变化。我们最近 建立了一种新的无血清纯化培养人星形胶质细胞的方法。使用我们的新 方法，我们将对从患有糖尿病的人类患者中纯化的反应性星形胶质细胞进行分子表征。 癫痫、脑肿瘤、阿尔茨海默病、帕金森病和动静脉畸形。目标1： 将表征反应性星形胶质细胞的转录组，并测试分子表型 反应性星形胶质细胞在人类中是多样的。在初步研究中，我们发现， 状态，根据疾病状况，星形胶质细胞有不同的反应状态。我们会研究 使用人星形胶质细胞的体外培养物在人中的反应性星形胶质细胞中诱导的分子的功能。在 目的2，我们将直接比较人类和小鼠星形胶质细胞对各种有害刺激的反应。 我们的初步数据表明，人类和小鼠星形胶质细胞对氧化应激的敏感性不同 有害刺激激活了人类和小鼠星形胶质细胞中不同的信号通路。这些研究 有可能揭示反应性星形胶质细胞在人类神经系统疾病中的作用或失败， 为治疗癫痫、脑肿瘤和神经退行性疾病提供了新的治疗靶点。

Project Summary/Abstract Astrocytes constitute at least one third of all cells in the human brain and are critical for the development and function of the central nervous system. Reactive astrogliosis is a spectrum of cellular, molecular, and functional changes of astrocytes found in a wide range of injuries and diseases, including epilepsy, brain tumor, Alzheimer’s disease, Parkinson’s disease, stroke, inflammation, and traumatic brain injuries. Based on studies of mouse models, reactive astrocytes play both beneficial and harmful roles in disease progression and neural repair by secreting cytokines that regulate immune cells, producing growth factors, and forming scars that insulate disease tissue from healthy tissue. However, little is known about the molecular and cellular changes of astrocytes in human patients, due in part to the difficulties of purifying and culturing human astrocytes. Previous methods of purifying human astrocytes rely on serum, which induces reactive astrogliosis in the purification procedure, making it difficult to investigate reactive changes of astrocytes in patients. We recently developed a novel purification and culturing method for human astrocytes without serum. Using our new method, we will perform molecular characterization of reactive astrocytes purified from human patients with epilepsy, brain tumor, Alzheimer’s disease, Parkinson’s disease, and arteriovenous malformation. In Aim 1, we will characterize the transcriptome of reactive astrocytes and test the hypothesis that the molecular phenotypes of reactive astrocytes are diverse in humans. In preliminary studies, we found that instead of being a single state, there are diverse reactive states of astrocytes depending on the disease condition. We will examine the function of molecules induced in reactive astrocytes in humans using in vitro cultures of human astrocytes. In Aim 2, we will directly compare the responses of human and mouse astrocytes to a variety of harmful stimuli. Our preliminary data showed that human and mouse astrocytes have different susceptibility to oxidative stress and that harmful stimulus activates different signaling pathways in human vs. mouse astrocytes. These studies has the potential to reveal what reactive astrocytes do or fail to do in human neurological disorders, and provide new therapeutic targets for treating epilepsy, brain tumor, and neurodegenerative disorders.