4-Phenylbutyrate (4-PBA) has been shown to restore dysferlin localization and activity in cell culture models of dysferlinopathy, a muscular dystrophy caused by DYSF missense mutations. To support therapeutic testing, cell culture models were optimized using myoblasts derived from mice lacking the dysferlin protein and HEK cells transfected with human dysferlin mutations. A flow cytometry based assay was used to quantify dysferlin relative to wild-type controls. Treatment with 4-PBA significantly increased dysferlin membrane localization in mouse myoblasts and V67D HEK cells. The L1341P mutation was also examined in a downstream analysis of skeletal muscle histology from the L1341P mouse model to assess disease progression. H&E-stained myofibers were generated through a collaborative study and the proportion of centrally nucleated myofibers was quantified as a marker of muscle regeneration. Female mice treated with 4-PBA exhibited higher muscle weights and evidence of slower disease progression compared to untreated controls. 

In parallel, a novel cell model was developed for alpha-sarcoglycanopathy, a muscular dystrophy caused by SGCA missense mutations. HEK cells were transfected with human SGCA mutations and a flow cytometry based membrane localization assay was established to quantify SGCA expression. 4-PBA produced modest restoration of SGCA localization in mutant cells, revealing its broader effects on membrane associated proteins. Although 4-PBA consistently increased dysferlin localization in multiple models, its mechanism of action remains unclear. Ongoing work focuses on dysferlin structure and drug–protein interactions to identify how 4-PBA rescues function, thereby supporting its potential as a therapeutic and advancing cell based models for drug discovery.