Study of cell-type specific Alzheimer's disease genetic variants using a novel bioengineered model of iPSC-derived neural tissue

使用 iPSC 衍生神经组织的新型生物工程模型研究细胞类型特异性阿尔茨海默病遗传变异

• 批准号: 9980545
• 负责人: PHILIP G HAYDON
• 金额: $ 97.59万
• 依托单位: TUFTS UNIVERSITY BOSTON
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-05-01 至 2025-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/9980545
• 关键词: 3-Dimensional; Abeta clearance; Action Potentials; Affect; Aging; Alleles; Alzheimer like pathology; Alzheimer' s Disease; Alzheimer' s disease brain; Alzheimer' s disease pathology; Alzheimer' s disease patient; Alzheimer' s disease risk; Amyloid beta-42; Amyloid beta-Protein; Animal Disease Models; Animal Model; Apolipoprotein E; Apolipoproteins; Architecture; Astrocytes; Axon; Binding; Biomedical Engineering; Brain; Cell Culture Techniques; Cell Density; Cell Line; Cell Surface Proteins; Cell Transplantation; Cell model; Cells; Cholesterol Homeostasis; Code; Cognitive; Collagen; Complement; Complex; Data; Deterioration; Diffusion; Electrophysiology (science); Family; Functional disorder; Gene Expression; Genes; Genetic Polymorphism; Genotype; Growth; Homeostasis; Human; In Vitro; Late Onset Alzheimer Disease; Learning; Link; Mammals; Memory; Memory impairment; Metabolic; Microglia; Modeling; Mus; Mutation; Myeloid Cells; Necrosis; Neonatal; Nerve Degeneration; Neurodegenerative Disorders; Neuroglia; Neurons; Nutrient; Optics; Oxygen; Pathogenesis; Pathologic; Patients; Phagocytosis; Phenotype; Physiological; Population; Porifera; Proteins; Risk; Risk Factors; Rodent; Senile Plaques; Silk; Sleep; Synapses; Synaptic Transmission; Synaptic plasticity; System; TREM2 gene; Technology; Time; Tissue Model; Tissues; Transplant Recipients; Transplantation; Variant; abeta deposition; apolipoprotein E-4; base; beta secretase; brain behavior; cell type; early onset; exome; extracellular; familial Alzheimer disease; genetic variant; genome editing; genome sequencing; genome wide association study; human subject; in vivo; induced pluripotent stem cell; innovation; lipid metabolism; member; molecular phenotype; mutant; neuronal excitability; novel; precursor cell; receptor; relating to nervous system; risk variant; scaffold; stem cell technology; synaptic function; synaptogenesis; three dimensional cell culture; trafficking; whole genome

ABSTRACT Alzheimer’s disease (AD) is a progressive neurodegenerative disorder characterized by memory impairments and cognitive deterioration. Aging is the major risk factor for AD. Furthermore, increasing evidence indicates that astrocytes and microglia are implicated in the pathogenesis of AD. The ε4 allele of the apolipoprotein E gene (APOE) has been identified as a major risk factor contributing to the pathogenesis of sporadic AD (SAD) in about 15-20% of the cases. APOE is the major apolipoprotein expressed in the human brain primarily by astrocytes and to a lesser extent by microglia, and is involved in cholesterol homeostasis, and regulates A clearance. Furthermore, genome-wide association studies (GWAS) have identified polymorphisms in genes enriched in microglia (e.g. SORL1, CR1, CD2AP, CD33, TREM2, ABCA7) and astrocytes (e.g. CLU and ABCA7) that increase the risk of developing AD. Recent advances in stem cell technology have allowed the reprogramming of primary cells from human subjects into induced pluripotent stem cells (iPSCs) and their differentiation in neurons, astrocytes and microglia. However, conventional 2D culture systems fail to recapitulate the diversity and maturation of multiple cell types and their interaction under physiological and pathological conditions. To overcome these weaknesses we have developed a novel bioengineered model of iPSC-derived neural tissue. Our silk-collagen protein-based ‘donut’ scaffolds can support compartmentalized, 3D brain-like tissues over a year, without necrosis. This tissue model is highly innovative, supporting the differentiating neurons growth in a donut-shaped porous silk sponge within an optically cleared collagen-filled central region for axon connectivity and synapse formation, that will allow for the first time live in vivo studies (e.g., cell-based electrophysiology, trafficking, synaptic functionality) of an human AD brain-like tissue during ageing (months of cultivation) under controlled experimental conditions. More importantly, the architecture of the scaffolds was optimized to meet the metabolic demand of high-density cell cultures in terms of free diffusion of nutrients and oxygen, a fundamental requisite for long-term cultures and ageing-related studies. Thus, we propose to: 1) Assess genotype-phenotype relationship of AD genetic variants enriched in astrocytes and microglia in patient-derived 3D brain-like cultures; 2) Assess genotype-phenotype relationship of AD genetic variants in vivo after transplantation of patient-derived cells in mice.

抽象的 阿尔茨海默氏病（AD）是一种以记忆障碍为特征的进行性神经退行性疾病 认知恶化。 星形胶质细胞和小胶质细胞与AD的发病机理有关。 基因（APOE）已被视为有助于零星AD发病的主要危险因素（SAD） 在大约15-20％的情况下。 星形胶质细胞和小胶质细胞的程度较小，参与胆固醇稳态，并调节A 清除。 富含小胶质细胞（例如SORL1，CR1，CD33，TREM2，ABCA7）和星形胶质细胞（例如CLU和 ABCA7）增加了干细胞技术最新进展的风险。 来自人类受试者的原代细胞的重射仪诱导多能干细胞（IPSC）和Theeir 但是，神经元，星形胶质细胞和小胶质细胞的区分。 概括多样性和成熟成熟的多种凯尔特人类型及其相互作用的脑相互作用和 为了克服这些弱点，我们已经开发了一种新型的生物工程模型 IPSC衍生的神经组织。 3D大脑样组织一年，没有坏死。 在甜甜圈形的多孔丝海绵中区分神经元的生长在光学清除的胶原蛋白 轴突连通性和突触形成的中央区域，这将首次允许体内研究 （例如，基于细胞的电生理学，运输，突触功能）在人类AD脑样组织中 在受控的实验条件下，老化（几个月的培养）。 优化了脚手架以满足自由培养的高密度培养的代谢需求 营养和氧的扩散，这是长期培养和与年龄相关的研究的基本必需品。 因此，我们建议：1）评估富含星形胶质细胞的AD遗传学变体的基因型 - 表型关系 与患者衍生的3D大脑样培养物中的小胶质细胞； 2） 在患者衍生细胞移植后体内遗传变异。