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 进行性瘫痪和死亡。假设 microRNA (miRNA) 进行转录后基因调控 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会受到破坏， 例如 TDP43、FUS 和 SMN。然而，特定 miRNA 在人类运动神经元基因调控中的作用和 功能没有得到很好的表征。我之前发现单个 miRNA，miR-218，是独特富集的 并在小鼠运动神经元中大量表达。此外，缺乏 miR-218 的小鼠在 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之间的关系 表型尚未解决，并且 miR-218 的生物学作用之前尚未在 人类，在我们对人类运动神经元基因调控的了解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人类 诱导多能干（hiPS）细胞衍生的人类运动神经元发育和功能模型，称为 皮质运动组合体，通过融合皮质、脊髓和骨骼肌球体。阿明博士建议使用 这个新颖的系统可以模拟 miR-218 对运动神经元发育、靶标通路和人类的影响 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 精通运动神经元发育、miRNA 生物学和高级转录组学，并将实现新的 与导师 Sergiu Pasca 博士一起接受干细胞生物学和人脑类器官模型的职业发展培训。 阿明博士将利用斯坦福大学卓越的研究环境和资源。他会 得到他的顾问委员会的支持，该委员会由非编码 RNA 介导的专家 Howard Chang 博士组成 基因调控，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）是一位执业神经科医生和疾病发病机制专家。该提案的完成将为 进一步研究 miR-218 及其靶标 mRNA 在人类运动中的治疗调节作用的方法 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.