Project ID CM-HD2023_55

ThemeCM-HD

Co Supervisor 1A Randall Centre for Cell and Molecular BiophysicsWebsite

Co Supervisor 1B Randall Centre for Cell and Molecular BiophysicsWebsite

Regenerative therapies for muscle-wasting diseases

Muscular dystrophies are a group of diseases that are characterised by muscle weakness, where muscle wastes and connective tissue becomes deregulated, so scar tissue forms. Oxidative stress from respiration is common to many muscular dystrophies. A common muscular dystrophy is Facioscapulohumeral (FSHD), caused by ectopic expression of the transcription factor DUX4, in which muscle cells are clearly sensitive to oxidative stress (doi: 10.15252/emmm.202013695). Treatment of patients with anti-oxidants vitamin E, vitamin C, zinc, and selenomethionine improves muscle function (clinicaltrials.gov:NCT01596803) (doi:10.1016/j.freeradbiomed.2014.09.014). We have found that mitochondrial biosynthesis/dysfunction and subsequent generation of oxidative stress contributes to FSHD pathology, which can be ameliorated by drugs/antioxidants (doi: 10.1093/hmg/ddy405, 10.1016/j.redox.2022.102251, Figure).

Aims:
To define interventions that will enhance mitochondrial function and increase protection against oxidative stress to improve muscle function.

Techniques and Skills:
Molecular Biology (e.g. cloning), Cell Biology (mouse/human cell culture, retroviral-transduction, siRNA-mediated gene-knockdown), Animal Models, Gene Expression/Protein Analysis (RT-qPCR, Western blotting, immunolabeling), Imaging/Time-Lapse microscopy, Metabolomics and Bioinformatics.

Objectives:
• Year 1: Better define mitochondrial function/oxidative stress in FSHD muscle/fibroblasts using metabolomics. Characterise connective tissue perturbation by analysing gene expression data from FSHD muscle biopsies.
• Year 2: Test selected drugs to improve mitochondrial function/reduce oxidative stress on FSHD patient cell lines and on muscle/connective tissue expressing DUX4.
• Year 3/4: Investigate interaction of drugs with known/novel signalling pathways to identify mechanisms. Test candidates in animal models of muscular dystrophy

Expertise:
• Zammit: muscle stem cell function in health and disease.
• Logan: muscle associated connective tissue and its disease associations.

One representative publication from each co-supervisor:

• Heher, P., Ganassi, M., Weidinger, A., Engquist, E.N., Pruller, J., Nguyen, T.H., Tassin, A., Declèves, A.E., Mamchaoui, K., Banerji, C.R.S., Grillari, J., Kozlov, A.V., and Zammit, P.S. (2022). Interplay between mitochondrial reactive oxygen species, oxidative stress and hypoxic adaptation in facioscapulohumeral muscular dystrophy: metabolic stress as potential therapeutic target. Redox Biol. 51, 102251 (doi: 10.1016/j.redox.2022.102251).

• Besse, L., Sheeba C.J., Holt M., Labuhn M., Wilde S., Feneck E., Bell D., Kucharska A., Logan M.P.O.(2020) Individual limb muscle bundles are formed through progressive steps orchestrated by adjacent connective tissue cells during primary myogenesis Cell Reports 30,(10), p.355-3665 DOI:10.1016/j.celrep.2020.02.037