Ballester López, AlfonsinaNúñez Manchón, JuditKoehorst, EmmaLinares Pardo, IanAlmendrote, MiriamLucente, GiuseppeGuanyabens, NicolauLópez Osías, MartaLucía Mulas, AlejandroNogales-Gadea, GiselaEt al.2020-10-172020-10-172020Ballester-López, A., Núñez-Manchón, J., Koehorst, E., Linares-Pardo, I., Almendrote, M., Lucente, G., Guanyabens, N., López-Osías, M., Suárez-Mesa, A., Hanick, S. A., Chojnacki, J., Lucía Mulas, A., Pintos-Morell, G., Coll-Cantí, J., Martínez-Piñeiro, A., Ramos-Fransi, A., & Nogales-Gadea, G. (2020). Three-dimensional imaging in myotonic dystrophy type 1: Linking molecular alterations with disease phenotype. Neurology Genetics, 6(4), e484. https://doi.org/10.1212/NXG.00000000000004842376-7839http://hdl.handle.net/11268/9157Objective: We aimed to determine whether 3D imaging reconstruction allows identifying molecular:clinical associations in myotonic dystrophy type 1 (DM1). Methods: We obtained myoblasts from 6 patients with DM1 and 6 controls. We measured cytosine-thymine-guanine (CTG) expansion and detected RNA foci and muscleblind like 1 (MBNL1) through 3D reconstruction. We studied dystrophia myotonica protein kinase (DMPK) expression and splicing alterations of MBNL1, insulin receptor, and sarcoplasmic reticulum Ca(2+)-ATPase 1. Results: Three-dimensional analysis showed that RNA foci (nuclear and/or cytoplasmic) were present in 45%-100% of DM1-derived myoblasts we studied (range: 0-6 foci per cell). RNA foci represented <0.6% of the total myoblast nuclear volume. CTG expansion size was associated with the number of RNA foci per myoblast (r = 0.876 [95% confidence interval 0.222-0.986]) as well as with the number of cytoplasmic RNA foci (r = 0.943 [0.559-0.994]). Although MBNL1 colocalized with RNA foci in all DM1 myoblast cell lines, colocalization only accounted for 1% of total MBNL1 expression, with the absence of DM1 alternative splicing patterns. The number of RNA foci was associated with DMPK expression (r = 0.967 [0.079-0.999]). On the other hand, the number of cytoplasmic RNA foci was correlated with the age at disease onset (r = -0.818 [-0.979 to 0.019]). Conclusions: CTG expansion size modulates RNA foci number in myoblasts derived from patients with DM1. MBNL1 sequestration plays only a minor role in the pathobiology of the disease in these cells. Higher number of cytoplasmic RNA foci is related to an early onset of the disease, a finding that should be corroborated in future studies.engAttribution-NonCommercial-NoDerivatives 4.0 Internacionalhttp://creativecommons.org/licenses/by-nc-nd/4.0/journal article10.1212/NXG.0000000000000484open accessGenética humanaTecnología médicaImágenes tridimensionales en medicinaGenética humanaTecnología médica