Transcriptome analysis of C2C12 myoblasts with mutations in LMNA gene

Mutations in LMNA gene cause laminopathies in human. Mostly, laminopathies alter skeletal muscle tissue and lead to cardiomathy and lipodystrophy. We investigated the effect of mutations G232E (EDMD2 syndome) and R482L (FPLD2 syndrome) in LMNA gene on skeletal muclse functioning and metabolism using transgenic C2C12 myoblast cell lines and transcriptome analysis. We found abnormalities of nuclear lamina structure in mutant myoblasts, treir pro-myogenic commitment and metabolic disregulation on all stages of transgenic myoblasts differentiation. C2C12 mouse myoblast cell line was transduced with lentivaral constraction with mutant human LMNA gene. We got three transgenic cell lines: WT-hLMNA C2C12, G232E-hLMNA C2C12 and R482L-hLMNA C2C12. These transgenic myoblasts were differentiated in myogenic direction (using differentiation medium with 2% horse serum). We collected RNA samples on day 0 (undifferentiated myoblasts, controls), day 2 and day 4 of differentiation for all cell lines. Samples were in triplicate. In total, there are 9 biological conditions (3 cell lines, 3 differantiation days) and 27 samples. Libraries were prepared with TruSeq Stranded mRNA kit, samples were RNA-sequenced on Illumina HiSeq 2500, read length 50 bp.