Modulating miR-218 in human motor neurons using assembloids

使用组合体调节人类运动神经元中的 miR-218

• 批准号: 10525638
• 负责人: Neal Dilip Amin
• 金额: $ 19.52万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-08-08 至 2027-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10525638
• 关键词: 3' Untranslated Regions; 3-Dimensional; ALS patients; Advisory Committees; Antibodies; Autocrine Communication; Autopsy; Award; Axon; Bioinformatics; Biological; Biological Assay; Biological Process; Biology; Brain; CRISPR/Cas technology; Cessation of life; DNA Sequence Alteration; Defect; Development; Disease; Disease Pathway; Dose; Environment; Evolution; Exhibits; Functional disorder; Future; Gene Expression; Gene Expression Regulation; Genes; Glutamates; Goals; Histology; Human; Image; Inherited; Investigation; Knowledge; Ligands; Mediating; Mediator of activation protein; Mentors; Messenger RNA; MicroRNAs; Modeling; Motor; Motor Neuron Disease; Motor Neurons; Mus; Muscle; Muscle Contraction; Mutation; Nervous system structure; Neurologist; Neuromuscular Junction; Neurons; Organoids; Paralysed; Pathogenesis; Pathway interactions; Phenotype; Post-Transcriptional Regulation; Proteins; Regulation; Regulator Genes; Regulatory Pathway; Reporter; Repression; Research; Resources; Role; Scientist; Skeletal Muscle; Spinal; Sum; Synapses; System; Therapeutic; Tissues; Training; Universities; Untranslated RNA; base; bioinformatics pipeline; career development; comparative; design; experience; expression vector; gene function; gene network; human model; induced pluripotent stem cell; insight; motor control; motor neuron development; motor neuron function; mouse genetics; neurodevelopment; neuromuscular; neuromuscular function; novel; optogenetics; overexpression; predictive modeling; presynaptic; protein TDP-43; receptor; response; skills; stem cell biology; stem cell model; synaptogenesis; transcriptome sequencing; transcriptomics

PROJECT SUMMARY/ABSTRACT In motor neuron diseases, neuromuscular junctions are lost and motor neurons degenerate resulting in progressive paralysis and death. Post-transcriptional gene regulation by microRNAs (miRNAs) is hypothesized to be disrupted in motor neuron diseases due to inherited mutations in proteins involved in miRNA processing, such as TDP43, FUS, and SMN. Yet, the role of specific miRNAs in human motor neuron gene regulation and function is not well characterized. I previously discovered that a single miRNA, miR-218, is uniquely enriched and abundantly expressed in mouse motor neurons. Furthermore, mice lacking miR-218 exhibited deficits in neuromuscular synaptogenesis and die due to muscle paralysis – phenotypes associated with motor neuron disease. Subsequent studies have implicated miR-218 dysregulation as a mediator of motor neuron disease in humans. However, the relationship between miR-218’s repression of target gene pathways and motor neuron phenotypes has not been resolved, and the biological role of miR-218 has not been previously investigated in humans, leaving an important translational gap in our knowledge of human motor neuron gene regulation and function. In response to this challenge, we in the Pasca Lab have recently developed a three-dimensional, human induced pluripotent stem (hiPS) cell-derived model of human motor neuron development and function, called cortico-motor assembloids, by fusing cortical, spinal, and skeletal muscle spheroids. Dr. Amin proposes using this novel system to model the impact of miR-218 upon motor neuron development, target pathways, and human specific-features of post-transcriptional gene regulation. This proposal will leverage Dr. Amin’s existing proficiencies in motor neuron development, miRNA biology, and advanced transcriptomics and will enable new career development training in stem cell biology and human brain organoid models with mentor Dr. Sergiu Pasca. Dr. Amin will utilize the exceptional research environment and resources available at Stanford University. He will be supported by his advisory committee comprising of Dr. Howard Chang, an expert in non-coding RNA mediated gene regulation, Dr. Aaron Gitler, an expert in motor neuron biology and disease pathways, and Dr. Richard Reimer, a practicing neurologist and expert in disease pathogenesis. Completion of this proposal will pave the way for further investigations into the therapeutic modulation of miR-218 and its target mRNAs in human motor neuron disease.

项目摘要/摘要 在运动神经元疾病中，神经肌肉连接丢失，运动神经元变性导致 渐进的瘫痪和死亡。假设通过microRNA（miRNA）调节转录后基因 由于参与miRNA加工的蛋白质的遗传突变，在运动神经元疾病中受到破坏， 例如TDP43，FUS和SMN。然而，特定miRNA在人体运动神经元基因调节中的作用和 功能没有很好地表征。我以前发现单个miRNA miR-218是独特的富集 并在小鼠运动神经元中绝对表达。此外，缺乏miR-218暴露不足的小鼠 神经肌肉突触发生并因肌肉麻痹而死亡 - 与运动神经元相关的表型 疾病。随后的研究已实施miR-218失调作为运动神经元疾病的介体 人类。但是，miR-218的反射靶基因途径与运动神经元之间的关系 表型尚未得到解决，MiR-218的生物学作用以前尚未在 人类，在我们对人类运动神经元基因调节和 功能。为了应对这一挑战，我们在PASCA实验室中已经开发了一个三维的人类 诱导人类运动神经元发育和功能的诱导多能茎（臀部）细胞衍生的模型称为 皮质运动组件，通过融合皮质，脊柱和骨骼肌球体。 Amin博士使用 这个新型系统，用于建模miR-218对运动神经元发展，靶途径和人类的影响 转录后基因调控的特定功能。该建议将利用Amin博士的现有 运动神经元发展，miRNA生物学和高级转录组学的熟练程度，将启用新的 通过心理塞尔吉·帕斯卡（Sergiu Pasca）博士，干细胞生物学和人脑器官模型的职业发展培训。 阿明博士将利用斯坦福大学提供的卓越研究环境和资源。他会的 他的咨询委员会符合他的咨询委员会的支持，霍华德·张博士（Howard Chang）是非编码RNA介导的专家 基因调节，运动神经元生物学和疾病途径专家Aaron Gitler博士和Richard博士 雷默（Reimer），疾病发病机理的实践神经科医生和专家。该提案的完成将铺平 进一步研究MiR-218的治疗调节及其靶标mRNA的方法 神经元疾病。