Abstract Details

Title JAG2 Variant Cloning: Evaluation of a Muscular Dystrophy Gene and a Potential Model System

Topic Neuromuscular and Clinical Neurophysiology (EMG)

Presentation(s) P3 - Poster Session 3 (5:30 PM-6:30 PM)

Poster/Presentation
Number 11-001

Background The JAG2 gene encodes the protein Jagged2, an integral ligand in the evolutionarily conserved Notch signaling pathway. Notch signaling is critical in directing myriad developmental processes, thus pathogenic variants that compromise these pathways result in a set of serious inherited diseases. Biallelic pathogenic variants in JAG2 have recently been associated with muscular dystrophy. In the aftermath of this discovery, questions have arisen regarding the regulatory environment of JAG2 in healthy and diseased skeletal muscle and how potential therapies can be developed, necessitating the development of a suitable model system.

Objective

To discern whether expressing human JAG2 pathogenic variants in myoblast cultures will generate a reliable model system for muscular dystrophy.

Design/Methods

To explore the potential of C2C12 mouse myoblasts as an in vitro model system we developed a framework wherein both transfected and non-transfected cells were studied concurrently. We synthesized plasmids representing known JAG2 pathogenic variants and transfected them into shRNA scrambled control cells and shRNA JAG2 knockdown cells. Simultaneously, identical studies were carried out on shRNA control cells and shRNA-mediated knockdown cells devoid of any plasmid transfection. Both sets of cells underwent identical assessments of proliferation, differentiation, and downstream molecular characteristics.

Results

We successfully constructed and transfected 3 plasmids representing severe, moderate, and mild JAG2 variants as well as empty vector and JAG2 reference sequence configurations resulting in 10 stable cell lines representing the 5 plasmids transfected into scrambled control and JAG2 shRNA cells. Significant differences in cellular growth rates between cell lines transfected with different plasmids were observed, providing initial insights into the potential role of JAG2 mutations in cellular behavior.

Conclusions We anticipate that our ongoing studies will illuminate the effects of specific pathogenic variants in JAG2 on skeletal muscle development in this model system, setting the stage for testing both molecular and small molecule therapies.

Authors/Disclosures
Johnnie Turner PRESENTER	The institution of Mr. Turner has received research support from American Academy of Neurology.
Hannah Littel	No disclosure on file
MEKALA GUNASEKARAN (UNIVERSITY OF MINNESOTA)	No disclosure on file
Peter B. Kang, MD, FAAN (University of Minnesota Medical School)	Dr. Kang has received personal compensation in the range of $10,000-$49,999 for serving as a Consultant for Novartis. Dr. Kang has received personal compensation in the range of $5,000-$9,999 for serving as a Consultant for Sarepta Therapeutics. Dr. Kang has received personal compensation in the range of $500-$4,999 for serving as a Consultant for Neurogene. Dr. Kang has received personal compensation in the range of $500-$4,999 for serving as a Consultant for NS Pharma. Dr. Kang has received personal compensation in the range of $500-$4,999 for serving as a Consultant for Teneofour. Dr. Kang has received personal compensation in the range of $500-$4,999 for serving as a Consultant for Lupin. Dr. Kang has received personal compensation in the range of $10,000-$49,999 for serving as an Editor, Associate Editor, or Editorial Advisory Board Member for Wiley. The institution of Dr. Kang has received research support from Xtraordinary Joy Foundation. The institution of an immediate family member of Dr. Kang has received research support from Sarepta Therapeutics. The institution of Dr. Kang has received research support from Centers for Disease Control and Prevention (CDC). The institution of Dr. Kang has received research support from National Institutes of Health (NIH). The institution of Dr. Kang has received research support from Food and Drug Administration. The institution of Dr. Kang has received research support from National Initiative for Cockayne Syndrome. The institution of Dr. Kang has received research support from Greg Marzolf Jr Foundation. An immediate family member of Dr. Kang has received intellectual property interests from a discovery or technology relating to health care. Dr. Kang has received publishing royalties from a publication relating to health care. Dr. Kang has received personal compensation in the range of $5,000-$9,999 for serving as a Medical Advisory Board with National Initiative for Cockayne Syndrome. Dr. Kang has a non-compensated relationship as a Medical Advisory Board with Speak Foundation that is relevant to AAN interests or activities. Dr. Kang has a non-compensated relationship as a TRIAD Council with Hereditary Neuropathy Foundation that is relevant to AAN interests or activities. Dr. Kang has a non-compensated relationship as a President with Child Neurology Society that is relevant to AAN interests or activities. Dr. Kang has a non-compensated relationship as a Board Member with Child Neurology Foundation that is relevant to AAN interests or activities.