| Autor: |
Robinson KG; Nemours Children's Research, Nemours Children's Health System, Wilmington, DE 19803, USA., Marsh AG; Center for Bioinformatics and Computational Biology, University of Delaware, Newark, DE 19716, USA., Lee SK; Nemours Children's Research, Nemours Children's Health System, Wilmington, DE 19803, USA., Hicks J; Center for Bioinformatics and Computational Biology, University of Delaware, Newark, DE 19716, USA., Romero B; Medical and Molecular Sciences, University of Delaware, Newark, DE 19716, USA., Batish M; Medical and Molecular Sciences, University of Delaware, Newark, DE 19716, USA., Crowgey EL; Nemours Children's Research, Nemours Children's Health System, Wilmington, DE 19803, USA., Shrader MW; Department of Orthopedics, Nemours Children's Hospital Delaware, Wilmington, DE 19803, USA., Akins RE; Nemours Children's Research, Nemours Children's Health System, Wilmington, DE 19803, USA. |
| Abstrakt: |
Spastic type cerebral palsy (CP) is a complex neuromuscular disorder that involves altered skeletal muscle microanatomy and growth, but little is known about the mechanisms contributing to muscle pathophysiology and dysfunction. Traditional genomic approaches have provided limited insight regarding disease onset and severity, but recent epigenomic studies indicate that DNA methylation patterns can be altered in CP. Here, we examined whether a diagnosis of spastic CP is associated with intrinsic DNA methylation differences in myoblasts and myotubes derived from muscle resident stem cell populations (satellite cells; SCs). Twelve subjects were enrolled (6 CP; 6 control) with informed consent/assent. Skeletal muscle biopsies were obtained during orthopedic surgeries, and SCs were isolated and cultured to establish patient-specific myoblast cell lines capable of proliferation and differentiation in culture. DNA methylation analyses indicated significant differences at 525 individual CpG sites in proliferating SC-derived myoblasts (MB) and 1774 CpG sites in differentiating SC-derived myotubes (MT). Of these, 79 CpG sites were common in both culture types. The distribution of differentially methylated 1 Mbp chromosomal segments indicated distinct regional hypo- and hyper-methylation patterns, and significant enrichment of differentially methylated sites on chromosomes 12, 13, 14, 15, 18, and 20. Average methylation load across 2000 bp regions flanking transcriptional start sites was significantly different in 3 genes in MBs, and 10 genes in MTs. SC derived MBs isolated from study participants with spastic CP exhibited fundamental differences in DNA methylation compared to controls at multiple levels of organization that may reveal new targets for studies of mechanisms contributing to muscle dysregulation in spastic CP. |
