B37Investigation Of Viability And Response To Inflammatory Stimuli In Cultured Human Myotubes Derived From Patients With Huntington's Disease

• Published in: Journal of neurology, neurosurgery and psychiatry Vol. 85; no. Suppl 1; p. A22
• Main Authors: Parker, JA, Haider, S, Miller, JRC, Brown, S, Robertson, N, Lewis, M, Sillery, E, Nowak, V, Sethi, H, Bjorkqvist, M, Orth, M, Tabrizi, S J
• Format: Journal Article
• Language: English
• Published: 01-09-2014
• ISSN: 0022-3050
• DOI: 10.1136/jnnp-2014-309032.65

BackgroundMuscle atrophy has been well documented in patients with HD and yet its cause is unknown. The presence of a low grade immune response in HD has been implicated in muscle wasting. Myoblasts and myotubes within skeletal muscle are known not only to respond to inflammatory cytokines including IL-6 and TNF alpha , but also release cytokines. The presence of mutant HTT within the myoblasts and myotubes of HD patients may affect the release of cytokines from these cells as has previously been observed in HD monocytes and macrophages.AimsTo assess the viability and response to inflammatory stimuli in human HD myotubes in vitro.MethodsAs part of the Multi-Tissue Molecular signatures in HD project (MTM-HD) we have collected muscle biopsies from early stage HD patients and matched controls. Myoblast cultures have been established from these biopsies and differentiated into myotubes. Differentiation has been assessed using immunofluorescent staining for myoblast and myotube markers. Release of IL-6 has been measured using an electrochemiluminescent assay.ResultsThe myoblast content and differentiation potential of these cultures from control and early HD patients have been determined and compared. The release of IL-6 in response to lipopolysaccharide (LPS), TNF alpha and IL-6 by myoblasts and myotubes from control and early HD patients was measured. As HD is an age-related disease, the effects of ageing myotubes in vitroon their viability and response to LPS have also been assessed.ConclusionsHere data is presented comparing the differentiation of HD and control myoblasts, their survival in culture and response to inflammatory stimuli.