SBIR-STTR Award

Defective membrane repair is associated with the progression of muscular dystrophy that is linked to mutations in caveolin-3 (Cav3) and dysferlin in human patients. Several other forms of muscular dystrophy, including Duchenne muscular dystrophy and dystrophy arising from mutations in the dystroglycan complex have been linked to membrane fragility. Compromised membrane repair and increased membrane fragility are distinct mechanisms leading to increased muscle fiber death, as evidenced by the additive nature of these two pathways. A therapeutic approach to increase the capacity of muscle cells to reseal their membranes following physiological levels of mechanical stress could address both of these mechanisms leading to improvement of the regenerative capacity in muscular dystrophy. Attempts to produce therapeutics targeting membrane resealing have been complicated by the lack of knowledge of the molecular components involved. Recent studies show that Mitsuguimin 53 (MG53), a muscle-specific TRIM-family protein (TRIM72), is an essential component of the acute membrane repair machinery. MG53 acts as a sensor of oxidation to nucleate recruitment of intracellular vesicles to the injury site for membrane patch formation. MG53 can interact with dysferlin to facilitate its membrane repair function, and the membrane trafficking function of MG53 can be modulated through a functional interaction with Cav3. Our data indicate that a molecular complex formed by MG53, dysferlin and Cav3 is essential for repair of muscle membrane damage, thus providing a therapeutic target for treatment of muscular and cardiovascular diseases. In an effort to translate these basic science findings into therapeutic interventions for human diseases, we have formed a biotechnology company named TRIM-edicine, Inc, based on intellectual properties discovered in at UMDNJ-Robert Wood Johnson Medical School. Our research and development effort at TRIM-edicine has provided extensive studies to show that recombinant MG53 purified from E. coli retains efficient membrane repair function, supporting the therapeutic value of targeting MG53 in muscular dystrophy and other human diseases. We have preliminary in vivo animal model data to show that intra-muscular delivery of recombinant MG53 can ameliorate cardiotoxin-induced damage to the muscle fibers. This project will comprise an effort by TRIM-edicine that will leverage our expertise to pursue the proof-of-principle studies for the therapeutic application of MG53 in treatment and/or prevention of various types of muscular dystrophy. , ,

Public Health Relevance:
Muscular dystrophies are a family of genetic disorders that all generally include progressive muscle weakness due to degeneration of the muscle fibers, which includes the most common inherited disease, Duchene Muscular dystrophy. Many of these diseases involve either fragility of the membranes that surround muscle cells or a compromised ability to reseal those membranes. Both of these cases lead to compromised integrity of the cell membrane that result in death of muscle fibers, eventual depletion of the muscle regenerative capacity, muscle fibrosis, decreased force production and in many cases death of the patient. If a therapeutic approach could address membrane fragility and reduced resealing capacity it would have efficacy across a large number of different muscular dystrophies. Current efforts within the regenerative medicine field involve examining ways to increase muscle repair in syndromes where there is a reduced regenerative capacity for the skeletal muscle. In this project, TRIM-edicine will leverage our expertise to pursue proof-of- principle studies for the therapeutic application of a novel protein, mitsugumin 53 (MG53) in treatment and/or prevention of various types of muscular dystrophy.

Thesaurus Terms:
A/J Mouse;Acute;Address;Animal Model;Animal Models And Related Studies;Basic Research;Basic Science;Biosynthetic Proteins;Biotechnology;Blocking Antibodies;Blood Serum;Cardiotoxin;Cardiovascular Diseases;Causality;Cell Communication And Signaling;Cell Signaling;Cell Membrane;Cellular Injury;Cessation Of Life;Cobra Cardiotoxin;Complex;Cranin;Cytoplasmic Membrane;Dysf Protein, Human;Data;Death;Development And Research;Direct Lytic Factors;Disease;Disorder;Drug Kinetics;Duchene;Duchenne;Duchenne De Boulogne Muscular Dystrophy;Duchenne Disease;Duchenne Dystrophy;Duchenne Muscular Dystrophy;Duchenne Muscular Dystrophy (Dmd);Duchenne Myodystrophy;Duchenne Pseudohypertrophic Muscular Dystrophy;Duchenne Syndrome;Duchenne-Griesinger Syndrome;Dystroglycan;Dystroglycans;Dystrophin;E Coli;Ellis-Van Creveld (Evc) Syndrome;Escherichia Coli;Etiology;Exposure To;Extracellular Space;Fermentation;Fibrosis;Future;Generations;Genetic Alteration;Genetic Change;Genetic Condition;Genetic Diseases;Genetic Engineering Of Proteins;Genetic Defect;Goals;Hereditary;Hereditary Disease;Human;Human, General;Inbred Mdx Mice;Inherited;Injury;Intellectual Property;Intercellular Space;Intracellular Communication And Signaling;Investigation;Knowledge;Laboratories;Lead;Limb-Girdle Muscular Dystrophies;Link;M-Caveolin;Man (Taxonomy);Man, Modern;Mechanical Stress;Membrane;Membrane Protein Traffic;Membrane Traffic;Mice, Inbred Mdx;Modeling;Molecular;Molecular Disease;Mouse, Mdx;Movement;Muscle;Muscle Cells;Muscle Cells, Mature;Muscle Fibers;Muscle Tissue;Muscle Weakness;Muscle, Skeletal;Muscle, Voluntary;Muscular Dystrophies;Muscular Dystrophies, Limb-Girdle;Muscular Dystrophy, Duchenne;Muscular Dystrophy, Pseudohypertrophic;Muscular Weakness;Mutation;Myocytes;Myodystrophica;Myodystrophy;Myotubes;Names;Nature;Pathology;Pathway Interactions;Patients;Pb Element;Pharmacokinetics;Phase;Physiologic;Physiological;Plasma Membrane;Preparation;Prevention;Principal Investigator;Production;Progressive Muscular Dystrophy, Duchenne Type;Property;Property, Loinc Axis 2;Protein Engineering;Proteins;Protocol;Protocols Documentation;Pseudohypertrophic Muscular Dystrophy, Childhood;R & D;R&D;Recombinant Proteins;Recombinants;Regenerative Medicine;Rhabdomyocyte;Rodent Model;Sbir;Sbirs (R43/44);Serum;Severities;Signal Transduction;Signal Transduction Systems;Signaling;Site;Skeletal Fiber;Skeletal Muscle Cell;Skeletal Muscle Fiber;Skeletal Muscle Tissue;Skeletal Myocytes;Skeletal Muscle Structure;Small Business Innovation Research;Small Business Innovation Research Grant;Syndrome;Trim Family;Therapeutic;Therapeutic Intervention;Therapeutic Studies;Therapy Research;Translating;Translatings;Vescl;Vesicle;Wood;Wood Material;X-Linked Dilated Cardiomyopathy;X-Linked Dilated Cardiomyopathy (Xlcm);X-Linked Muscular Dystrophy;X-Linked Recessive Muscular Dystrophy;Base;Benign X-Linked Recessive Muscular Dystrophy;Biological Signal Transduction;Body Movement;Cardiovascular Disorder;Caveolin-3;Cell Damage;Cell Injury;Childhood Pseudohypertrophic Muscular Dystrophy;Classic X-Linked Recessive Muscular Dystrophy;Disease Causation;Disease Etiology;Disease/Disorder;Disease/Disorder Etiology;Disorder Etiology;Dysferlin;Dysferlin, Limb Girdle Muscular Dystrophy 2b (Autosomal Recessive) Protein, Human;Family Genetics;Gene Product;Genetic Disorder;Genome Mutation;Heavy Metal Pb;Heavy Metal Lead;Hereditary Disorder;Human Dysf Protein;Human Disease;In Vivo;Intervention Therapy;Language Translation;Limb-Girdle Muscular Weakness And Atrophy;Limb-Girdle Syndrome;Medical Schools;Membrane Structure;Mild X-Linked Recessive Muscular Dystrophy;Model Organism;Muscular Dystrophy Mouse Model;Myopathic Limb-Girdle Syndrome;Novel;Oxidation;Pathway;Phase 1 Study;Plasmalemma;Progressive Muscular Dystrophy Of Childhood;Pseudohypertrophic Adult Muscular Dystrophy;Pseudohypertrophic Muscular Paralysis;Pseudohypertrophic Progressive Muscular Dystrophy, Duchenne Type;Public Health Relevance;Regenerative;Repair;Repaired;Research And Development;Restoration;Safety Testing;Sensor;Therapeutic Protein;Therapeutic Target

This application requests for a Phase II project for our initial SBIR grant addressing the use of a novel protein therapeutic agent, MG53, for the treatment of muscular dystrophy (1R43AR060019). Our Phase I studies established manufacturing and formulation conditions, and provided significant proof- of-concept data for the use of MG53 in treating muscular dystrophy. This Phase II application proposed additional proof-of concept experiments in multiple dystrophy models (mouse, dog) and critical toxicology studies. This proposal will directly target the muscular dystrophies, a family of geneti disorders that generally include progressive muscle weakness due to degeneration of the muscle fibers that is linked to either fragility of the membranes that surround muscle cells or a compromised ability to reseal those membranes. An emerging concept in recent biomedical research establishes that intrinsic membrane repair/regeneration is a fundamental aspect of normal physiology and that disruption of this repair function underlies the progression of many human diseases, including muscular dystrophy. A therapeutic approach to increase the capacity of muscle cells to reseal their membranes following physiological levels of mechanical stress could address both of these mechanisms leading to improvement of the regenerative capacity in muscular dystrophy. Attempts to produce therapeutics targeting this unmet medical need have been complicated by the lack of knowledge of the molecular components involved. Recent studies show that MG53, a muscle-specific TRIM-family protein, plays an essential role in protection of skeletal and cardiac muscle cells against various types of acute injury or chronic physiological stresses. MG53 ablation results in defective sarcolemmal membrane repair with progressive muscle pathology. In an effort to translate these basic science findings into therapeutic interventions for human diseases, we have formed a biotechnology company named TRIM-edicine, Inc, based on intellectual properties discovered at UMDNJ-Robert Wood Johnson Medical School. Our research and development effort at TRIM-edicine during Phase I of this SBIR project has established methods to produce large quantities of recombinant MG53 protein purified from E. coli that retains efficient membrane repair function. Furthermore, we produced in vivo animal model data using dystrophic mdx mice to show that intravenous (IV) delivery of recombinant MG53 can repair membrane damage to skeletal muscle and ameliorate the pathology associated with muscular dystrophy. Additional in vivo data show that repetitive IV-injections of recombinant human MG53 to mice are safe and do not produce adverse effects. Other studies showed that subcutaneous injection of MG53 can be effective for therapeutic approaches as well. This project will leverage the expertise at TRIM-edicine in biologic drug development to pursue necessary studies for development of recombinant MG53 in treatment of muscular dystrophy to allow for the filing of an Investigational New Drug (IND) application with the Food and Drug Administration (FDA). This Phase II continuation project will involve a collaborative effort between TRIM-edicine and the University of North Carolina to pursue the proof-of-principle studies for the therapeutic application of MG53 in treatment of muscular dystrophy using the golden retriever model of muscular dystrophy. First, we will produce sufficient amount of recombinant dog MG53 proteins following our documented Chemical Manufacture Control (CMC) protocols. Second, we will test the efficacy and safety of the dog MG53 protein in reducing muscular pathology using a golden retriever with muscular dystrophy (GRMD) dog model. Third, upon completion of the in vivo animal model studies, the selection of animal species for the toxicology studies will be determined upon consultation with FDA in a pre-IND meeting. Completion of the safety/toxicology evaluations will provide the final step in preparation for filing of an IND application testing the use of recombinant MG53 in treatment of muscular dystrophy in human patients.

Public Health Relevance:
SBIR 1R43AR060019 Phase II application: ""Protein Therapeutics for Muscular Dystrophy"" Principal Investigator: Takizawa, Norio

Project narrative:
Muscular dystrophies are severe genetic disorders that are associated with defects in muscle cell membrane integrity or a reduced capacity for muscle to repair damage that occurs during normal contraction. Attempts to produce therapeutics targeting this unmet medical need have been complicated by the lack of knowledge of the basic biology of cell membrane repair. The discovery of MG53 as a key component of the muscle membrane repair machinery has opened a new therapeutic approach for treatment of muscular dystrophy. The development effort proposed in this project will generate the proof-of principle data and the safety evaluation for using recombinant MG53 protein as a therapeutic agent for treatment of muscular dystrophy.