Muscle degeneration underlies many forms of muscular dystrophy and may also contribute significantly to the decline of muscle mass associated with the normal aging process. Limb Girdle Muscular Dystrophy 2B is a recessive form of muscular dystrophy that is due to mutations in the dysferlin gene. Dysferlin is member of the ferlin family. In muscle, two different ferlin proteins are known to mediate membrane fusion events. Muscle growth arises because myoblasts fuse to each other as well as to the multinucleate syncytial myofiber. In addition, upon membrane damage, fusion of intracellular vesicles to the plasma membrane facilitates the rapid resealing of membrane disruption. Dysferlin is more abundantly expressed in mature myofibers, and muscle lacking dysferlin displays ineffective membrane sealing of laser-induced membrane disruption. Myoferlin is highly expressed in myoblasts undergoing fusion to myotubes, and myoferlin null myoblasts fuse less effectively forming smaller myotubes in vivo and in vitro. Both dysferlin and myoferlin have at least six C2 domains, and these domains regulate membrane fusion in other C2-domain containing proteins. The first C2 domains of both dysferlin and myoferlin bind to negatively charged phospholipids only in the presence of calcium. These data reinforce the role of ferlin proteins as scaffolds for assembling and mediating the fusion events important for muscle growth and repair. Insulin-like growth factor (IGF) 1 is a compound known to stimulate muscle growth, and we now found that myoferlin cells are unresponsive IGF1. Additionally, we found that IGF receptors do not translocate normally in myoferlin null myoblasts. In a separate line of experiments, we found that the second C2 domain of myoferlin binds directly to EHD2;EHD proteins have been implicated in endocytic recycling, a process where internalized vesicular components fuse to the plasma membrane to return cargo to the plasma membrane. We propose a broad role for ferlin proteins in vesicle trafficking that includes endocytosis and recycling defects. This vesicle traffic is critical for normal myoblast fusion and muscle growth and repair.
Aim 1 will examine the role ferlin proteins, specifically myoferlin, in receptor trafficking and signaling.
Aim 2 will identify and study proteins associated with endocytic recycling to determine their role in myoblast fusion and muscle growth. Understanding this pathway will guide the design of new therapies to promote muscle growth and repair in disease and aging.

Public Health Relevance

Ferlin proteins are important for muscle growth and repair. Mutations in dysferlin cause inherited muscular dystrophy, a devastating progressive muscle disorders that causes the loss of walking and breathing. Myoferlin, is highly related to dysferlin, and is important for normal muscle growth. We are studying the role of ferlin proteins to devise therapy to improve muscle growth in the muscular dystrophies.

Agency
National Institute of Health (NIH)
Institute
National Institute of Neurological Disorders and Stroke (NINDS)
Type
Research Project (R01)
Project #
2R01NS047726-05A2
Application #
7783103
Study Section
Skeletal Muscle and Exercise Physiology Study Section (SMEP)
Program Officer
Porter, John D
Project Start
2003-07-01
Project End
2011-08-31
Budget Start
2009-09-30
Budget End
2010-08-31
Support Year
5
Fiscal Year
2009
Total Cost
$390,000
Indirect Cost
Name
University of Chicago
Department
Internal Medicine/Medicine
Type
Schools of Medicine
DUNS #
005421136
City
Chicago
State
IL
Country
United States
Zip Code
60637
Quattrocelli, Mattia; Capote, Joanna; Ohiri, Joyce C et al. (2017) Genetic modifiers of muscular dystrophy act on sarcolemmal resealing and recovery from injury. PLoS Genet 13:e1007070
Quattrocelli, Mattia; Barefield, David Y; Warner, James L et al. (2017) Intermittent glucocorticoid steroid dosing enhances muscle repair without eliciting muscle atrophy. J Clin Invest 127:2418-2432
Demonbreun, Alexis R; McNally, Elizabeth M (2017) Muscle cell communication in development and repair. Curr Opin Pharmacol 34:7-14
Quattrocelli, Mattia; Salamone, Isabella M; Page, Patrick G et al. (2017) Intermittent Glucocorticoid Dosing Improves Muscle Repair and Function in Mice with Limb-Girdle Muscular Dystrophy. Am J Pathol 187:2520-2535
Demonbreun, Alexis R; McNally, Elizabeth M (2016) Plasma Membrane Repair in Health and Disease. Curr Top Membr 77:67-96
Demonbreun, Alexis R; Quattrocelli, Mattia; Barefield, David Y et al. (2016) An actin-dependent annexin complex mediates plasma membrane repair in muscle. J Cell Biol 213:705-18
Lamar, Kay-Marie; Miller, Tamari; Dellefave-Castillo, Lisa et al. (2016) Genotype-Specific Interaction of Latent TGF? Binding Protein 4 with TGF?. PLoS One 11:e0150358
Demonbreun, Alexis R; Allen, Madison V; Warner, James L et al. (2016) Enhanced Muscular Dystrophy from Loss of Dysferlin Is Accompanied by Impaired Annexin A6 Translocation after Sarcolemmal Disruption. Am J Pathol 186:1610-22
Lenhart, Kaitlin C; O'Neill 4th, Thomas J; Cheng, Zhaokang et al. (2015) GRAF1 deficiency blunts sarcolemmal injury repair and exacerbates cardiac and skeletal muscle pathology in dystrophin-deficient mice. Skelet Muscle 5:27
Demonbreun, Alexis R; McNally, Elizabeth M (2015) DNA Electroporation, Isolation and Imaging of Myofibers. J Vis Exp :e53551

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