Comparison of Wild Type & Mutated Dysferlin Muscle Cells

Abstract

Dysferlinopathy is a rare form of muscular dystrophy caused by a mutation of the dysferlin gene. The mutation causes loss of membrane repair in skeletal muscle cells. A patient diagnosed with dysferlinopathy will experience progressive muscle deterioration resulting in loss of function of the lower limbs. Although patients live an average lifespan, there is no cure for the disease and they become wheelchair dependent. To understand the workings of the disease, the experiment manipulated mouse myoblasts: C2C12 cells as a control line and GREG cells to mimic the effects of dysferlinopathy. To ensure the absence of dysferlin in GREG, the mutation in the DYSF gene was confirmed by PCR. Differentiation of the myoblasts into myotubes had been used to compare the phenotypes of C2C12 and GREG. When differentiation began, the mutated cells formed myotubes significantly faster, longer in length, and thinner than wild type myoblasts. Almost every mutant myoblast transformed into a myotube whereas the wildtype myoblasts had a heterogenous population of myoblasts and myotubes. Implication of these results could be that the wild type cells are better at maintaining their stem cell population. When there is damage to the mutated cell membrane due to muscle contraction, there may be an insufficient amount of stem cells available to repair the affected area. One possible course of action could be gene therapy via transfection. To test this, we will attempt to transfect wild type DYSF into the mutated cell line and determine if this rescues the rapid differentiation.

Jumily Bourommavong | Biology and Chemistry | Faculty Sponsor Eric Williams