Exosome-mediated propagation of disease linked poly-dipeptides in C9orf72-FTD/ALS

C9orf72-FTD/ALS 中疾病相关多二肽的外泌体介导的传播

基本信息

批准号：
9425328
负责人：
Davide Trotti
金额：
$ 356.01万
依托单位：
THOMAS JEFFERSON UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-08-01 至 2023-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9425328
关键词：
Amyotrophic Lateral Sclerosis Area Behavioral Biochemical Biogenesis Biological Assay Biological Markers Biological Process Brain Brain Diseases C9ORF72 Cell Culture Techniques Cells Cerebrospinal Fluid Clinical Communication Complex Dipeptides Disease Ectopic Expression Enterobacteria phage P1 Cre recombinase Fluorescent Antibody Technique Frontotemporal Dementia Genes Genetic Image Image Analysis Imaging Techniques Impaired cognition In Vitro Injections Injury Label Link Lipids Mediating Messenger RNA MicroRNAs Modality Monitor Motor Neurons Mus Nanotubes Nature Nerve Degeneration Neurodegenerative Disorders Neuroglia Neurons Nucleotides Pathogenesis Pathogenicity Pathway interactions Patients Production Proteins RNA Reporter Reporting Research Route Site Sorting - Cell Movement Spinal Symptoms Testing Time Toxic effect Transcript Transgenic Mice Translations Work base cell type exosome extracellular vesicles in vivo induced pluripotent stem cell intercellular communication live cell imaging migration neural circuit neuron loss neurotoxic novel novel strategies promoter protein TDP-43 tau Proteins transmission process uptake

项目摘要

A growing body of evidence uncovered a propensity for frontotemporal dementia (FTD) and amyotrophic lateral sclerosis (ALS) pathogenic proteins to propagate from cell-to-cell. Although few other mechanisms have been proposed, secretion of exosomes has been reported to occur from different neural cell types, including neurons, and to potentially serve as a new intercellular communication route within the CNS. Interestingly, based on the evidence of focality and neuroanatomical propagation of clinical symptoms, it was also hypothesized that the cerebro-spinal fluid (CSF) could serve as vehicle for pathogenic proteins spread, at least in ALS. Utilizing different in vitro cell culture platforms, including spinal motor neurons derived from iPSCs of C9orf72 patients, we recently learnt that C9orf72-linked dipeptide proteins (DPRs) spread between neural cells via the exosome-dependent pathway. By analyzing a newly generated exosome-reporter transgenic mouse, we also found that exosomes are capable of migrating extensive distance in vivo. These observations led us to postulate that an exosome-mediated propagation of DPRs could be a modality by which toxic insults spread in disease-afflicted CNS areas in C9orf72-FTD/ALS. We will be testing using complementary in vitro and in vivo approaches the novel hypothesis that transmitted DPRs transfer injury via exosomes to both neighboring cells, but also to neurons downstream in synaptic circuits. We propose: (1) To investigate exosome-mediated mechanisms of DPRs transmission in CNS cells; (2) To examine the modalities of cell-to-cell propagation of DPRs in vivo; (3) To examine whether cell transfer of DPRs propagates toxicity. The proposed work has the potential to open up an entirely new field of C9orf72 FTD/ALS research, at the same time, providing important clues to the fundamental biological processes in brain cellular communications relevant to brain diseases. Thus, the results are expected to have a significant impact for understanding C9orf72-linked FTD/ALS pathogenesis and eventually treating patients.

越来越多的证据发现了额颞痴呆（FTD）和肌萎缩性侧索硬化症（ALS）致病蛋白从细胞到细胞传播。虽然很少已经提出了机制，据报道外泌体的分泌是从不同神经出发的细胞类型，包括神经元，并有可能充当新的细胞间通信路线中枢神经系统。有趣的是，基于临床的焦点和神经解剖传播的证据症状，还假设脑脊髓液（CSF）可以用作车辆致病蛋白至少在ALS中传播。利用不同的体外细胞培养平台，包括衍生自IPSC的脊柱运动神经元在C9ORF72患者中，我们最近了解到C9ORF72连接的二肽蛋白（DPR）在神经细胞通过外泌体依赖性途径。通过分析新生成的外泌体养子转基因小鼠，我们还发现外泌体能够在体内迁移较长的距离。这些观察结果使我们假设外泌体介导的DPR传播可能是有毒损伤在C9ORF72-FTD/ALS中疾病伴的CNS地区传播的方式。我们会的使用互补的体外和体内进行测试，方法是传播DPRS的新假设通过外泌体将损伤转移到两个相邻细胞，也可以在突触回路下游的神经元中转移损伤。我们建议：（1）研究中枢神经系统细胞中DPRS传播的外泌体介导的机制；（2）检查体内DPR的细胞间传播的模态；（3）检查是否 DPRS的细胞转移传播毒性。拟议的工作有可能打开C9orf72 FTD/ALS的全新领域同时，研究为大脑的基本生物学过程提供了重要的线索与脑部疾病有关的细胞通信。因此，预计结果将具有显着了解C9ORF72连接的FTD/ALS发病机理的影响，并最终治疗患者。