Poster abstracts
Poster number 22 submitted by Gianni Giarrano
Donor Cell Identity Determines the Therapeutic Potency of Dysferlin-Loaded Extracellular Vesicles
Gianni N. Giarrano (Ohio State Biochemistry Program ), Mia Kordowski, Juan David Salazar-Gil, Carman Taylor, Vidhi Amin (Department of Biomedical Engineering, Ohio State University), Caden Islam, Daniel Velsquez-Ramrez, Natalia Higuita-Castro (Department of Biomedical Engineering, Ohio State University), Britani N. Blackstone, Heather M. Powell (Shriners Childrens Ohio), Noah Weisleder (Department of Molecular and Cellular Biochemistry, University of Kentucky College of Medicine)
Abstract:
Dysferlinopathies are degenerative muscle disorders caused by autosomal recessive mutations in the dysferlin (DYSF) gene, resulting in diminished expression or loss of functional dysferlin and impaired sarcolemmal repair. Dysferlin is enriched in cardiac and skeletal muscle, where it mediates Ca2+-dependent plasma membrane repair following mechanical injury. Loss of functional dysferlin results in a decrease in plasma membrane repair capacity. This leads to the onset of dysferlinopathy with clinical symptoms such as progressive muscle, weakness, elevated serum creatine kinase levels, chronic inflammation, and abnormal muscle morphology. Despite extensive efforts, there are currently no curative therapies, with clinical management limited to symptomatic intervention and supportive mobility aids to mitigate disease progression. Here, we evaluate engineered extracellular vesicles (EVs) as a novel non-viral nanocarrier platform for delivering dysferlin mRNA cargo to dysferlin-deficient patient-derived cells. EVs were engineered through transient overexpression of dysferlin in donor human fibroblasts or myoblasts, and their ability to deliver dysferlin transcripts and restore membrane repair function was assessed in vitro. Despite comparable uptake, myoblast-derived dysferlin-loaded EVs (DYSF EVs) delivered higher levels of dysferlin mRNA and restored membrane repair following ballistic and laser-induced injury. In contrast, fibroblast-derived EVs failed to rescue repair. Proteomic profiling revealed that myoblast-derived EVs are enriched in cytoskeletal, membrane-associated, and repair-related proteins, suggesting coordinated delivery of dysferlin transcripts alongside endogenous components of membrane repair machinery. Together, these findings identify donor cell identity as a critical determinant of engineered EV therapeutic potency and support muscle-lineage EVs as a promising biomaterial strategy for restoring membrane repair in dysferlinopathies.
Keywords: Plasma membrane repair, Dysferlin, Extracellular vesicles