Project Type
Poster
Publication Date
4-15-2022
Department or Program
Biological Sciences
College
College of Arts and Sciences
Faculty Mentor #1
Morriss, Ginny
Abstract
Myotonic dystrophy type 1 (DM1) is a multisystemic disease characterized by muscle weakening and wasting. DM1 is the most commonly diagnosed adult-onset muscular dystrophy, and skeletal muscle wasting is responsible for 60% of DM1 related mortality, although the mechanism responsible for the DM1 skeletal muscle wasting phenotype has not been concretely identified. DM1 is caused by expansion of CUG repeats in the 3′ untranslated region of the DMPK gene. Pervious research has suggested that myokines, signaling molecules typically involved in cell-cell communication, play a role in muscle wasting. Additionally, gene expression analysis of a mouse DM1 model identified elevated expression of specific myokines, including Cx3cl1. This project aimed to explore the role of myokines in DM1 related muscle wasting using a cell culture model. Using Lipofectamine 3000, the C2C12 mouse myoblast cell line was transfected with plasmids that express either human DMPK exons 11-15 including a 960 CUG repeat or a control plasmid that that express DMPK exons 11-15 with no CUG repeats. Differentiation was induced in half of the cells, and RNA was extracted from the cells at three timepoints: day 1, 3, and 5. Extracted RNA was assessed using qPCR to quantify Cx3cl1 expression. Unfortunately, the qPCR results did not indicate measurable expression of Cx3cl1, the primary target myokine of this project, in the transfected cells, despite expression of internal controls at all timepoints at comparable level. The immeasurable expression of Cx3cl1 could be due to the C2C12 cells’ inadequate expression of CUG repeats in the DMPK gene, as the cells potentially are not expressing long enough repeats to observe an effect. Further research that utilizes a different model, such as a mouse skeletal muscle or human myoblast cell culture model is required to determine the influence of myokines on DM1 pathology, as these models express repeat RNA for prolonged periods of time.