Assessing Membrane Repair Capacity in Myoblast Using Dysferlin-Dependent Dye Tagging Techniques 

Presenter: Emma Ashlee DaPonte

Faculty Sponsor: Eric Owen Williams

School: Fitchburg State University

Research Area: Biology

Session: Poster Session 2, 11:30 AM - 12:15 PM, Auditorium, A61

ABSTRACT

Dysferlinopathy is a rare form of muscular dystrophy characterized by mutations in the dysferlin gene causing the production of misfolded dysferlin protein. This can lead to impaired cell membrane repair in skeletal muscle cells, and progressive muscle atrophy. Currently, there are no effective treatments for dysferlinopathy. Our research aims to identify compounds that are already FDA approved and test their capabilities to restore dysferlin’s repair function. To create a model of dysferlinopathy patients muscle cells, mouse muscle stem cells were transfected with human dysferlin genes in order to produce various mutated human dysferlin protein models. We focused on a specific pathogenic mutation, G299R, and tested the effects of 4-phenylbutyrate (4-PBA), Tauroursodeoxycholic acid (TUDCA); and cystic fibrosis correctors to determine if these compounds were able to restore the key functions of the dysferlin protein. We generated promising preliminary data that these compounds can effectively aid in restoring some of dysferlin’s functions due to their chemical chaperoning activity, with 4-PBA consistently performing the best. This cell line's response to varying concentrations of 4-PBA was tested and quantified using antibody fluorescent staining paired with flow cytometry. Cells were also stained with immunofluorescent dyes to visualize membrane repair efficiency and protein expression at the cells membrane using fluorescent microscopy. Our findings showed that higher concentrations of 4-PBA led to an increase in membrane targeting and partially restored dysferlin's membrane repair function suggesting that 4-PBA may have therapeutic potential for certain dysferlinopathy genotypes.  


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