Characterization of RAN Translation and Antisense Transcription in Primary Cell Cultures of Patients with Myotonic Dystrophy Type 1

Journal of Clinical Medicine

Ana Pilar Gómez-Escribano¹  Rafael P. Vázquez-Manrique¹  Jakub Chojnacki²  Alicia Martínez-Piñeiro³  Mònica Suelves³  Gisela Nogales-Gadea³  Emma Koehorst³  Judit Núñez-Manchón³  Guillem Pintos-Morell³  Giuseppe Lucente³  Alfonsina Ballester-López³  Alba Ramos-Fransi³  Miriam Almendrote³  Andrea Arbex³  Jaume Coll-Cantí³ 

[1] Centre for Biomedical Network Research on Rare Diseases (CIBERER), Instituto de Salud Carlos III, 28029 Madrid, Spain;IrsiCaixa AIDS Research Institute, 08916 Badalona, Spain;Neuromuscular and Neuropediatric Research Group, Institut d’Investigació en Ciències de la Salut Germans Trias i Pujol (IGTP), Campus Can Ruti, Universitat Autònoma de Barcelona, 08916 Badalona, Spain;

关键词: RAN translation;    antisense transcription;    myotonic dystrophies;    primary cell cultures;    phenotypic modulators;   

DOI  :  10.3390/jcm10235520

来源: DOAJ

【 摘 要 】

Myotonic Dystrophy type 1 (DM1) is a muscular dystrophy with a multi-systemic nature. It was one of the first diseases in which repeat associated non-ATG (RAN) translation was described in 2011, but has not been further explored since. In order to enhance our knowledge of RAN translation in DM1, we decided to study the presence of DM1 antisense (DM1-AS) transcripts (the origin of the polyglutamine (polyGln) RAN protein) using RT-PCR and FISH, and that of RAN translation via immunoblotting and immunofluorescence in distinct DM1 primary cell cultures, e.g., myoblasts, skin fibroblasts and lymphoblastoids, from ten patients. DM1-AS transcripts were found in all DM1 cells, with a lower expression in patients compared to controls. Antisense RNA foci were found in the nuclei and cytoplasm of a subset of DM1 cells. The polyGln RAN protein was undetectable in all three cell types with both approaches. Immunoblots revealed a 42 kD polyGln containing protein, which was most likely the TATA-box-binding protein. Immunofluorescence revealed a cytoplasmic aggregate, which co-localized with the Golgi apparatus. Taken together, DM1-AS transcript levels were lower in patients compared to controls and a small portion of the transcripts included the expanded repeat. However, RAN translation was not present in patient-derived DM1 cells, or was in undetectable quantities for the available methods.

【 授权许可】

Unknown