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: 10399527
• 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/10399527
• 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; Induced pluripotent stem cell derived neurons; Late Onset Alzheimer Disease; Learning; Link; Mammals; Memory; Memory impairment; Metabolic; Microglia; Modeling; Mus; Mutation; Myeloid Cells; Necrosis; 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; neonatal mice; 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.

摘要