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A non-coding hexanucleotide repeat expansion in intron 1 of the C9orf72 gene is the most common cause of amyotrophic lateral sclerosis and frontotemporal dementia (C9ALS/FTD), however, the precise molecular mechanism by which the C9orf72 hexanucleotide repeat expansion directs C9ALS/FTD pathogenesis remains unclear. Here, we report a novel disease mechanism arising due to the interaction of C9ORF72 with the RAB7L1 GTPase to regulate vesicle trafficking. Endogenous interaction between C9ORF72 and RAB7L1 was confirmed in human SH-SY5Y neuroblastoma cells. The C9orf72 hexanucleotide repeat expansion led to haploinsufficiency resulting in severely defective intracellular and extracellular vesicle trafficking and a dysfunctional trans-Golgi network phenotype in patient-derived fibroblasts and induced pluripotent stem cell-derived motor neurons. Genetic ablation of RAB7L1or C9orf72 in SH-SY5Y cells recapitulated the findings in C9ALS/FTD fibroblasts and induced pluripotent stem cell neurons. When C9ORF72 was overexpressed or antisense oligonucleotides were targeted to the C9orf72 hexanucleotide repeat expansion to upregulate normal variant 1 transcript levels, the defective vesicle trafficking and dysfunctional trans-Golgi network phenotypes were reversed, suggesting that both loss- and gain-of-function mechanisms play a role in disease pathogenesis. In conclusion, we have identified a novel mechanism for C9ALS/FTD pathogenesis highlighting the molecular regulation of intracellular and extracellular vesicle trafficking as an important pathway in C9ALS/FTD pathogenesis.

Original publication

DOI

10.1093/brain/awx024

Type

Journal article

Journal

Brain

Publication Date

01/04/2017

Volume

140

Pages

887 - 897

Keywords

C9ALS/FTD, C9orf72, RAB7L1, extracellular vesicles, haploinsufficiency, Amyotrophic Lateral Sclerosis, Animals, Biological Transport, C9orf72 Protein, COS Cells, Cell Line, Chlorocebus aethiops, DNA Repeat Expansion, Fibroblasts, Frontotemporal Dementia, Humans, Introns, Motor Neurons, Neurons, Oligonucleotides, Antisense, Pedigree, Pluripotent Stem Cells, Proteins, rab1 GTP-Binding Proteins