期刊
NEUROPATHOLOGY AND APPLIED NEUROBIOLOGY
卷 40, 期 6, 页码 670-685出版社
WILEY
DOI: 10.1111/nan.12148
关键词
amyotrophic lateral sclerosis; microarray; mRNA; post mortem; splicing; TDP-43
资金
- Motor Neurone Disease Association/Medical Research Council [G0800380/1]
- Medical Research Council
- Wellcome Trust
- European Union
- European Community [259867]
- Motor Neurone Disease Association
- STRENGTH, an EU Joint Programme - Neurodegenerative Disease Research project
- United Kingdom, Medical Research Council (MRC)
- MND Association/MRC [MR/K003771/1]
- MRC [G0900652, MR/K003771/1, MR/L016451/1, G0800380, G1100540, G0400074, G0502157, MR/K000039/1] Funding Source: UKRI
- Medical Research Council [G0502157, G0400074, G0800380, MR/K000039/1, MR/K003771/1, G1100540, MR/L016451/1, G0900652] Funding Source: researchfish
- National Institute for Health Research [NF-SI-0512-10082] Funding Source: researchfish
Aims: Loss of nuclear TDP-43 characterizes sporadic and most familial forms of amyotrophic lateral sclerosis (ALS). TDP-43 (encoded by TARDBP) has multiple roles in RNA processing. We aimed to determine whether (1) RNA-splicing dysregulation is present in lower motor neurones in ALS and in a motor neurone-like cell model; and (2) TARDBP mutations (mtTARDBP) are associated with aberrant RNA splicing using patient-derived fibroblasts. Methods: Affymetrix exon arrays were used to study mRNA expression and splicing in lower motor neurones obtained by laser capture microdissection of autopsy tissue from individuals with sporadic ALS and TDP-43 proteinopathy. Findings were confirmed by quantitative reverse transcription-polymerase chain reaction (qRT-PCR) and in NSC34 motor neuronal cells following shRNA-mediated TDP-43 depletion. Exon arrays and immunohistochemistry were used to study mRNA splicing and TDP-43 expression in fibroblasts from patients with mtTARDBP-associated, sporadic and mutant SOD1-associated ALS. Results: We found altered expression of spliceosome components in motor neurones and wide-spread aberrations of mRNA splicing that specifically affected genes involved in ribonucleotide binding. This was confirmed in TDP-43-depleted NSC34 cells. Fibroblasts with mtTARDBP showed loss of nuclear TDP-43 protein and demonstrated similar changes in splicing and gene expression, which were not present in fibroblasts from patients with sporadic or SOD1-related ALS. Conclusion: Loss of nuclear TDP-43 is associated with RNA processing abnormalities in ALS motor neurones, patient-derived cells with mtTARDBP, and following artificial TDP-43 depletion, suggesting that splicing dysregulation directly contributes to disease pathogenesis. Key functional pathways affected include those central to RNA metabolism.
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