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: 10678680
• 负责人: Neal Dilip Amin
• 金额: $ 22.88万
• 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/10678680
• 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; Mentors; Messenger RNA; MicroRNAs; Modeling; Motor; Motor Neuron Disease; Motor Neurons; Mus; Muscle; Muscle Contraction; Mutation; Nervous System; Neurologist; Neuromuscular Junction; Neuronal Dysfunction; 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; Synapses; System; Therapeutic; Tissues; Training; Universities; Untranslated RNA; Vertebral column; bioinformatics pipeline; career development; comparative; derepression; design; experience; expression vector; gene function; gene network; human model; induced pluripotent stem cell; insight; motor control; motor neuron degeneration; 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 在运动神经元病中，神经肌肉连接丢失，运动神经元退化，导致 进行性瘫痪和死亡。MicroRNAs(MiRNAs)对转录后基因调控的假说 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受到干扰， 例如TDP43、FUS和SMN。然而，特定的miRNAs在人类运动神经元基因调控和表达中的作用 功能没有得到很好的描述。我之前发现，单个miRNA miR-218具有独特的富集性 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 神经肌肉突触发生和肌肉麻痹所致死亡--与运动神经元相关的表型 疾病。随后的研究表明miR-218异常是运动神经元疾病的一个中介。 人类。然而，miR-218‘S抑制靶基因通路与运动神经元的关系 表型尚未解决，miR-218的生物学作用以前也没有被研究过 人类，留下了一个重要的翻译空白，我们的知识，人类运动神经元基因调控和 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 诱导多能干细胞(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 这个模拟miR-218对运动神经元发育、靶通路和人类影响的新系统 转录后基因调控的特异性。这项提案将利用阿明博士现有的 精通运动神经元发育、miRNA生物学和高级转录学，并将使新的 与导师塞尔吉奥·帕斯卡博士一起进行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出类拔萃的研究环境和资源。他会的 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亚伦·吉特勒博士和理查德博士 雷默是一位执业神经学家和疾病发病机制方面的专家。这项提案的完成将为 MiR-218及其靶向mRNAs在人体运动中的治疗调控研究方向 神经元病。

项目成果

Motor neurons use push-pull signals to direct vascular remodeling critical for their connectivity.

• DOI: 10.1016/j.neuron.2022.09.021
• 发表时间: 2022-12-21
• 期刊: NEURON
• 影响因子: 16.2
• 作者: Martins, Luis F.;Brambilla, Ilaria;Motta, Alessia;de Pretis, Stefano;Bhat, Ganesh Parameshwar;Badaloni, Aurora;Malpighi, Chiara;Amin, Neal D.;Imai, Fumiyasu;Almeida, Ramiro D.;Yoshida, Yutaka;Pfaff, Samuel L.;Bonanomi, Dario
• 通讯作者: Bonanomi, Dario