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Related Experiment Videos

Gene transfer to muscle using herpes simplex virus-based vectors

J Huard1, D Krisky, T Oligino

  • 1Department of Orthopaedic Surgery, Children's Hospital of Pittsburgh, PA 15261, USA.

Neuromuscular Disorders : NMD
|July 1, 1997
PubMed
Summary

Herpes simplex virus type 1 (HSV-1) vectors show promise for Duchenne muscular dystrophy (DMD) gene therapy by delivering the dystrophin gene. Researchers are overcoming viral cytotoxicity and improving muscle fiber transduction for effective treatment.

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Area of Science:

  • * Gene Therapy
  • * Virology
  • * Molecular Biology

Background:

  • * Duchenne muscular dystrophy (DMD) requires restoring dystrophin protein in muscle cells.
  • * Herpes simplex virus type 1 (HSV-1) is a potential viral vector for gene delivery due to its large genome and efficient transduction capabilities.
  • * Previous HSV-1 vectors faced challenges with cytotoxicity and inconsistent muscle fiber targeting.

Purpose of the Study:

  • * To evaluate replication-defective HSV-1 vectors for reporter gene transfer and expression in mouse skeletal muscle.
  • * To develop improved HSV-1 vectors to overcome cytotoxicity and enhance dystrophin gene delivery for DMD.
  • * To investigate the impact of muscle fiber maturity and basal lamina on HSV-1 transduction efficiency.

Main Methods:

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  • * Construction and testing of replication-defective HSV-1 mutants, including those with deletions in immediate early genes (ICP4, ICP22, ICP27, UL41).
  • * In vitro studies on myoblasts and myotubes to assess gene expression.
  • * In vivo intramuscular inoculation of HSV-1 vectors in newborn and adult mice.
  • * Immunohistochemical analysis to evaluate viral spread and transduction in different muscle types and ages.

    • Main Results:

    • * HSV-1 efficiently transduced myoblasts, myotubes, and muscle fibers in vivo, particularly in newborn mice.
    • * Multiple deletions in immediate early genes significantly reduced HSV-1 cytotoxicity compared to earlier strains.
    • * Differential transduction efficiency was observed between newborn and adult muscle fibers, suggesting potential barriers in mature muscle.

    Conclusions:

    • * Replication-defective HSV-1 vectors are viable for reporter gene delivery to skeletal muscle.
    • * Modified HSV-1 vectors with reduced cytotoxicity show potential for DMD gene therapy.
    • * Further research is needed to optimize HSV-1 vector delivery to adult muscle fibers, addressing barriers like the basal lamina.