Human-iPSC derived neuromuscular junctions as a model for neuromuscular diseases.

人 iPSC 衍生的神经肌肉接头作为神经肌肉疾病的模型。

• 批准号: 10727888
• 负责人: Helen C Miranda
• 金额: $ 39万
• 依托单位: CASE WESTERN RESERVE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-08 至 2024-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10727888
• 关键词: Accounting; Acetylcholine; Address; Adult; Amyotrophic Lateral Sclerosis; Animal Disease Models; Animal Model; Biological; Biological Assay; Biological Models; Brain; C9ORF72; CRISPR correction; Cell Line; Cells; Central Nervous System; Characteristics; Chromosome 9; Clinical Trials; Communication; Complex; Denervation; Development; Disease; Disease model; Drug Screening; Evaluation; Foundations; Functional disorder; Genes; Genetic; Genotype; Goals; Human; In Vitro; Individual; Introns; Investigation; Measures; Modeling; Molecular; Morphology; Motor Neurons; Mus; Muscle; Muscle Contraction; Mutation; Neuromuscular Diseases; Neuromuscular Junction; Organism; Pathway interactions; Patients; Pharmaceutical Preparations; Phase; Phased Innovation Awards; Phenotype; Physiological; Positioning Attribute; Predisposition; Prevention; Process; Protocols documentation; Reporting; Reproducibility; Signal Transduction; Skeletal Muscle; Spinal Cord; Stem Cell Research; Synaptic Cleft; System; Testing; Therapeutic; Validation; axion; disease phenotype; effective therapy; experience; experimental study; familial amyotrophic lateral sclerosis; frontotemporal lobar dementia amyotrophic lateral sclerosis; high throughput screening; induced pluripotent stem cell; multi-electrode arrays; neuronal survival; novel; optogenetics; prevent; reduce symptoms; reinnervation; spinal and bulbar muscular atrophy; stem cell model; stem cells

Motor neurons carry electrical signals from the brain through the spinal cord to ultimately generate muscle contraction via the neuromuscular junction (NMJ). The mechanisms involved in initiating and maintaining proper communication between the central nervous system and muscles are incredibly complex, and damage in this communication is the cause of neuromuscular diseases (NMD), such as Amyotrophic Lateral Sclerosis (ALS) and Spinal and Bulbar Muscular Atrophy (SBMA). While these disorders are among the most common NMDs, there is currently no cure or effective treatment for them. The NMD field acknowledges that a substantial number of drugs found to alleviate symptoms in animal models have failed in clinical trials. Even though this highlights the importance of the development of humanized models, a caveat of converting studies into iPSC models is the focus on single cells in detriment of the complex systems of the adult organism. In the NMD field specifically, iPSC investigations have largely focused on addressing motor neuron phenotypes that would prevent their degeneration. Unfortunately, this approach is no longer sufficient, as prolonging motor neuron survival does not assure re-innervation, nor does it guarantee prevention of denervation. Therefore, it is crucial for therapeutic advancement in the NMD field that stem cell research needs to focus not only on identifying cell-specific targets but also on testing those targets on functional NMJ systems that comprise both iPSC-derived motor neurons and skeletal muscles. To achieve this goal, we have recently developed a 2D functional human NMJ system comprised of both iPSC-derived motor neurons and skeletal muscles. Our human iPSC-NMJ model is responsive to optogenetics and we are able to quantitatively measure NMJ function in a multi-electrode array system. Hence, in this R61/R33 IGNITE Phased Innovation Award System, we propose to leverage our newly developed NMJ system to scale functional and morphological assessment (Aims 1 and 2) and validate the system by assaying NMJ-specific dysfunction using two NMDs: SBMA and ALS (Aims 3 and 4). Our lab has previously established an iPSC model for SBMA (R01NS121374-01, K01NS116119-01) and has extensive experience modeling this disease. Additionally, we selected the iPSCs harboring G4C2 hexanucleotide repeat expansion on chromosome 9 within the first intron of C9ORF72, as it represents is the most common genetic contributor to frontotemporal dementia (FTD) and ALS, accounting for ~10% of all cases of those diseases. Thus, successful completion of this R61/R33 will establish and validate a novel model system to facilitate therapeutic discovery for NMDs.

运动神经元将来自大脑的电信号通过脊髓传递，最终产生肌肉