Modulating miR-218 in human motor neurons using assembloids

项目概要/摘要 在运动神经元疾病中，神经肌肉接头丢失并且运动神经元退化，导致神经肌肉损伤。 进行性瘫痪和死亡。转录后基因调控的microRNAs（miRNAs）的假设 在运动神经元疾病中由于参与miRNA加工的蛋白质的遗传突变而被破坏， 例如TDP 43、FUS和SMN。然而，特定的miRNAs在人类运动神经元基因调控中的作用， 功能没有得到很好的描述。我之前发现，一个单一的miRNA，miR-218，是独特的富集， 并在小鼠运动神经元中大量表达。此外，缺乏miR-218的小鼠表现出在以下方面的缺陷： 神经肌肉突触发生和由于肌肉麻痹而死亡-与运动神经元相关的表型 疾病随后的研究表明，miR-218失调是运动神经元疾病的介质， 人类然而，miR-218对靶基因通路的抑制与运动神经元的关系尚不清楚。 表型尚未解决，miR-218的生物学作用以前也没有研究过。 人类，在我们对人类运动神经元基因调控的认识中留下了重要的翻译空白， 功能为了应对这一挑战，我们帕斯卡实验室最近开发了一种三维的， 人类运动神经元发育和功能的诱导多能干（hiPS）细胞衍生模型，称为 通过融合皮质、脊髓和骨骼肌球状体形成皮质-运动球状体。阿明博士建议使用 这个新的系统模拟了miR-218对运动神经元发育、靶向通路和人类神经元发育的影响。 转录后基因调控的具体特征。该提案将利用阿明博士现有的 在运动神经元发育，miRNA生物学和先进的转录组学的专业，并将使新的 与导师Sergiu Pasca博士一起进行干细胞生物学和人脑类器官模型的职业发展培训。 博士阿明将利用特殊的研究环境和资源，在斯坦福大学。他将 他的顾问委员会由非编码RNA介导的专家霍华德·张博士组成， 基因调控，亚伦Gitler博士，在运动神经元生物学和疾病通路的专家，和理查德博士 他是一位执业神经学家和疾病发病机理专家。完成这一建议将铺平道路 进一步研究miR-218及其靶mRNA在人运动神经中的治疗调节的方法 神经元疾病。

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.