Myoblast fusion is a fundamental process for proper skeletal muscle formation during development and regeneration. Despite the importance of myoblast fusion for muscle formation, the mechanisms that govern this process are not fully understood. Elucidation of fusion mechanisms is a critical step for understanding muscle development and to develop new therapeutic strategies to augment skeletal muscle disease. We recently discovered a muscle-specific protein, named myomaker, that localizes to the plasma membrane of myoblasts. Genetic deletion of myomaker during development and adult regeneration renders myoblasts fusion incompetent, which results in a dramatic inability to form skeletal muscle. Moreover, expression of myomaker in cells that normally do not express this protein causes their fusion with muscle cells. While myomaker is a central component for myoblast fusion, the regulatory mechanisms that govern its activity and the biochemical function of myomaker remain unknown. We have identified the regions of myomaker that are critical for its function and the proteins that interact with myomaker to potentially regulate its activity. In his project we will: 1) understand the function of myomaker within the fusion process 2) identify effectors of myomaker activity 3) evaluate the optimal cell type for heterologous fusion and myogenic reprogramming. We will use genetic loss-of- and gain-of-function approaches in vivo to investigate the role of myomaker during fusion. Extensive molecular approaches within our myomaker-based heterologous fusion system are proposed to identify the mechanisms that govern myomaker action. The ability of myomaker to drive fusion of heterologous cells with muscle cells will serve as the foundation to repair diseased skeletal muscle. We will use cell transplantation experiments to assess fusion and subsequent reprogramming of non-muscle cells with normal and diseased muscle. Successful completion of these aims will provide insight into the mechanisms of myoblast fusion by revealing how myomaker is regulated to induce membrane merger. Finally, these studies will have significant implications for a myomaker-mediated cell based strategy to rescue genetic muscle diseases.

Public Health Relevance

Fusion of myoblasts is an essential process for muscle development and regeneration. The goal of this project is to explore the mechanisms of myomaker action during myoblast fusion and to use that knowledge to develop new treatment strategies for skeletal muscle diseases.

Agency
National Institute of Health (NIH)
Institute
National Institute of Arthritis and Musculoskeletal and Skin Diseases (NIAMS)
Type
Research Project (R01)
Project #
5R01AR068286-03
Application #
9300865
Study Section
Skeletal Muscle and Exercise Physiology Study Section (SMEP)
Program Officer
Boyce, Amanda T
Project Start
2015-07-01
Project End
2020-06-30
Budget Start
2017-07-01
Budget End
2018-06-30
Support Year
3
Fiscal Year
2017
Total Cost
Indirect Cost
Name
Cincinnati Children's Hospital Medical Center
Department
Type
DUNS #
071284913
City
Cincinnati
State
OH
Country
United States
Zip Code
45229
Leikina, Evgenia; Gamage, Dilani G; Prasad, Vikram et al. (2018) Myomaker and Myomerger Work Independently to Control Distinct Steps of Membrane Remodeling during Myoblast Fusion. Dev Cell 46:767-780.e7
Sampath, Srihari C; Sampath, Srinath C; Millay, Douglas P (2018) Myoblast fusion confusion: the resolution begins. Skelet Muscle 8:3
Gamage, Dilani G; Leikina, Eugenia; Quinn, Malgorzata E et al. (2017) Insights into the localization and function of myomaker during myoblast fusion. J Biol Chem 292:17272-17289
Quinn, Malgorzata E; Goh, Qingnian; Kurosaka, Mitsutoshi et al. (2017) Myomerger induces fusion of non-fusogenic cells and is required for skeletal muscle development. Nat Commun 8:15665
Mitani, Yasuyuki; Vagnozzi, Ronald J; Millay, Douglas P (2017) In vivo myomaker-mediated heterologous fusion and nuclear reprogramming. FASEB J 31:400-411
Goh, Qingnian; Millay, Douglas P (2017) Requirement of myomaker-mediated stem cell fusion for skeletal muscle hypertrophy. Elife 6:
Millay, Douglas P; Gamage, Dilani G; Quinn, Malgorzata E et al. (2016) Structure-function analysis of myomaker domains required for myoblast fusion. Proc Natl Acad Sci U S A 113:2116-21