Development of marmoset models of neurodegenerative disease using embryonic stem cell-based gene-editing approaches

使用基于胚胎干细胞的基因编辑方法开发狨猴神经退行性疾病模型

• 批准号: 9209904
• 负责人: KUO-FEN LEE
• 金额: $ 74.21万
• 依托单位: SALK INSTITUTE FOR BIOLOGICAL STUDIES
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-04-01 至 2021-01-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9209904
• 关键词: Abeta synthesis; Age; Alleles; Alzheimer' s Disease; Alzheimer' s disease model; Amyloid beta-Protein; Amyloid beta-Protein Precursor; Anatomy; Animal Model; Animals; Area; Autistic Disorder; Autologous; Behavior; Behavioral; Brain; CRISPR/Cas technology; Callithrix; Cell Therapy; Cerebral cortex; Choline O-Acetyltransferase; Chronic Kidney Failure; Clinical; Clinical Trials; Cognitive; Degenerative Disorder; Dementia; Development; Diabetes Mellitus; Disease; Disease model; Embryo; Enterobacteria phage P1 Cre recombinase; Ethics; Exhibits; Functional disorder; Gene Proteins; Gene Targeting; Genes; Genetic; Genomics; Goals; Haplorhini; Hereditary Disease; Human; Human Amyloid Precursor Protein; Imagery; Impaired cognition; Institutes; Internal Ribosome Entry Site; Interneurons; Knock-in; Lead; Learning; Light; Longevity; Macaca mulatta; Maintenance; Malignant Neoplasms; Mediating; Medical Research; Methodology; Methods; Modeling; Modification; Morphology; Mosaicism; Mus; Mutation; National Institute of Drug Abuse; National Institute of General Medical Sciences; National Institute of Mental Health; National Institute of Neurological Disorders and Stroke; National Institute on Alcohol Abuse and Alcoholism; Nerve Degeneration; Neurobiology; Neurodegenerative Disorders; Neurologic; Neurons; Parkinson Disease; Parvalbumins; Pharmaceutical Preparations; Physiological; Point Mutation; Prevalence; Primates; Production; Protein Overexpression; Research; Research Personnel; Research Support; Resources; Schizophrenia; Stem cells; Stroke; Symptoms; System; Technology; Testing; Thinness; United States National Institutes of Health; abeta deposition; age related; basal forebrain; base; blastocyst; cartilaginous; cell type; cholinergic; cholinergic neuron; efficacy testing; embryonic stem cell; genetic manipulation; genome editing; hearing impairment; homologous recombination; human disease; improved; insight; mouse genome; mouse model; muscle form; nerve supply; neural circuit; neurogenesis; neuromechanism; neuronal circuitry; neuropathology; nonhuman primate; novel therapeutic intervention; pluripotency; promoter; protein expression; regenerative; regenerative therapy; screening; social; stem cell technology; tool

PROJECT SUMMARY The long-term goal of this project is to model human neurodegenerative diseases in marmosets via gene- editing in embryonic stem cells (ESCs). The mouse system is a powerful tool for medical research due to the ability to manipulate the mouse genome. However, considerable anatomical, physiological, cognitive, and behavioral differences between mice and humans limit the degree to which insights from mouse models shed light on human diseases. This is reflected in the high number of failed clinical trails for drugs that were effective in treating mouse models of human disease. Several lines of evidence suggest that the marmoset represents an improved animal system for studying a range of human diseases, including stroke and age-associated neurodegenerative diseases such as Alzheimer's disease (AD). Marmosets are the shortest-lived of the anthropoid primates (average lifespan of 5–7 years compared with 25 years for the rhesus macaque) and exhibit age-related changes that are similar to those seen in humans, including β-amyloid deposition in the cerebral cortex, loss of cholinergic innervation, and reduced neurogenesis, as observed in AD. In addition, marmosets are highly social and communicative and have demonstrated the capacity to learn sophisticated cognitive behaviors. Therefore, marmosets represent an ideal genetic platform for generating models of neurodegenerative diseases that more accurately reflect the human condition and enable the testing of potential autologous (the-same-species) stem cell-based regenerative therapies. Initial efforts will focus on generating a marmoset model of AD. The recent emergence of gene-editing and stem-cell technologies in primates pave the way toward generating marmoset disease models, but improvements in both areas are necessary to make this approach viable. Here, both conventional homologous recombination and CRISPR/Cas9 genome-editing technologies will be employed to modify marmoset ESCs. As genetic evidence demonstrates that mutations in the amyloid precursor protein (APP) gene result in increased β-amyloid production, the formation of plaques, and cognitive impairment, the marmoset APP will be edited to carry human point mutations. Genetic tools for studying neuronal cell type-specific circuits underlying cognitive impairment and neuropathology in AD will also be generated by inserting a Cre recombinase cassette into 3' end non-translated regions of the parvalbumin and choline acetyltransferase genes. These Cre driver lines will enable the visualization and functional manipulation of these cell types. Successful completion of the proposed Aims will generate a greatly improved animal model of AD, enable testing of stem cell-based regenerative methods for treating AD, and pave the way toward applying these genetic tools for analyzing neuronal circuitry of healthy brains. Establishing gene-editing in marmoset ESCs will also enable the development of additional primate models of human diseases, providing critical experimental resources for research supported by multiple NIH Institutes (e.g., NINDS, NIA, NIMH, NEI, NIAAA, NIDA, NICHG, NIGMS).

项目总结