Developing astrocyte, neuron, and microglial 3D organoids to model key aspects of human pathology

开发星形胶质细胞、神经元和小胶质细胞 3D 类器官来模拟人类病理学的关键方面

• 批准号: 9899912
• 负责人: Erik M Ullian
• 金额: $ 16.13万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-04-01 至 2021-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9899912
• 关键词: 3-Dimensional; Affect; Aging; Alzheimer' s Disease; Alzheimer' s disease risk; Astrocytes; Biological; Biology; Blood flow; Brain; Buffers; CRISPR interference; Cell physiology; Cells; Clustered Regularly Interspaced Short Palindromic Repeats; Complement; Complement 4b; Complex; Dementia; Developmental Biology; Disease; Drug Targeting; Foundations; Future; GRN gene; Gene Activation; Gene Expression; Genes; Goals; Homeostasis; Human; Human Pathology; Immune; Ions; Knock-out; Lead; Link; Lobular; Measures; Metabolic; Microglia; Modeling; Molecular; Morphology; Mus; Mutation; Nerve Degeneration; Neurodegenerative Disorders; Neuroglia; Neuronal Ceroid-Lipofuscinosis; Neurons; Neurotransmitters; Nuclear; Organoids; PGRN gene; Pathology; Pathway interactions; Patients; Phenotype; Play; Proteins; Research; Rodent; Rodent Model; Role; Signal Pathway; Signal Transduction; Stress; Synapses; Synaptic Transmission; Synaptic plasticity; System; TDP-43 aggregation; Temporal Lobe; Testing; Work; age related neurodegeneration; base; cell type; complement pathway; experimental study; extracellular; gene complementation; genome wide association study; glial activation; granulin; healthy aging; hippocampal sclerosis; human model; induced pluripotent stem cell; insight; interest; knock-down; neuron loss; neuronal survival; neurotoxic; novel; overexpression; prevent; protein TDP-43; protein aggregation; three dimensional cell culture; tool; transcription factor; translational study

Project Summary Astrocytes are the most abundant non-neuronal cell types in the brain. Astrocytes promote neuronal survival, provide neurons with metabolic substrates, help recycle synaptic neurotransmitters, regulate blood flow, buffer extracellular ions, induce or eliminate synapses and regulate synaptic transmission and synaptic plasticity. Most recently, it has been shown that when specific signaling pathways are compromised, astrocytes can become reactive and can start eliminating synapses ultimately; leading to neuronal cell death during both natural aging and age-related neurodegenerative diseases. Our research aims to understand how astroglial activation is initiated in Alzheimer’s Disease (AD). We will focus on this question by studying a gene called granulin (GRN). This gene has been linked to aging in the human cortex and loss of GRN inevitably leads to astroglial activation and a devastating neurodegenerative disease termed Frontal Temporal Lobar Degeneration (FTLD). To begin to tackle this question, we propose investigating the molecular mechanisms of GRN signaling on astroglial activation. The goal of our work is to understand how normal GRN function prevents protein mislocalization in healthy aging human neurons, and how loss of GRN causes these same proteins to mislocalize in neurons: leading to neuronal death in AD and FTLD. Based on recent findings that rodent astrocytes and human astrocytes are genetically and morphologically very different, we will use human cells to study this question and set up a novel 3D human induced pluripotent stem cell (iPSC) organoid system to incorporate human neurons, astrocytes and microglia. We will then analyze the mislocalization of specific protein in GRN positive and GRN knockout organoids. We will also study the signaling differences between GRN positive and GRN knockout astrocytes, and focus on the innate immune complement pathway and how GRN affects this pathway. Collectively, these studies will allow us to identify the factors in astrocytes that promote brain homeostasis, aging and disease.

项目摘要 星形胶质细胞是脑中最丰富的非神经元细胞类型。星形胶质细胞促进神经元存活， 为神经元提供代谢底物，帮助回收突触神经递质，调节血流，缓冲 细胞外离子，诱导或消除突触并调节突触传递和突触可塑性。 最近的研究表明，当特定的信号通路受损时，星形胶质细胞可以 变得反应性，并最终开始消除突触;导致神经元细胞死亡， 自然衰老和与年龄相关的神经退行性疾病。我们的研究旨在了解星形胶质细胞 激活在阿尔茨海默病（AD）中开始。我们将通过研究一种叫做 颗粒蛋白（GRN）。这种基因与人类大脑皮层的衰老有关，GRN的缺失不可避免地导致 星形胶质细胞激活和一种称为额颞叶的破坏性神经退行性疾病 变性（FTLD）。为了开始解决这个问题，我们建议调查的分子机制， GRN信号对星形胶质细胞活化的影响。我们工作的目标是了解正常的GRN功能 防止蛋白质在健康老化的人类神经元中的错误定位，以及GRN的丢失如何导致这些相同的 蛋白质在神经元中错误定位：导致AD和FTLD中的神经元死亡。根据最近的发现， 啮齿动物星形胶质细胞和人类星形胶质细胞在遗传和形态上非常不同，我们将使用人类星形胶质细胞。 细胞来研究这个问题，并建立一个新的3D人类诱导多能干细胞（iPSC）类器官系统 来整合人类神经元、星形胶质细胞和小胶质细胞。然后，我们将分析特定的错误定位 GRN阳性和GRN敲除类器官中的蛋白质。我们还将研究 GRN阳性和GRN敲除星形胶质细胞，并专注于先天免疫补体途径以及如何 GRN影响这一通路。总的来说，这些研究将使我们能够识别星形胶质细胞中的因子， 促进大脑内环境稳定，老化和疾病。