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Antibiotic-mediated selection of randomly mutagenized and cytokine-expressing oncolytic viruses

Reza Rezaei1,2, Stephen Boulton1, Mahsa Ahmadi3,4

  • 1Ottawa Hospital Research Institute, Ottawa, Ontario, Canada.

Nature Biomedical Engineering
|November 28, 2024
PubMed

Insights

This study introduces a novel antibiotic-based strategy to rapidly engineer oncolytic viruses. This method accelerates the development of enhanced cancer-fighting viruses by simplifying genome modification and gene deletion processes.

Area of Science:

  • Virology
  • Molecular Biology
  • Cancer Therapy

Background:

  • Therapeutic oncolytic viruses require genome engineering for enhanced efficacy and safety.
  • Current methods for generating recombinant viruses are complex and time-consuming, hindering development.

Purpose of the Study:

  • To develop an accelerated strategy for engineering oncolytic viruses.
  • To insert immunomodulatory transgenes and identify dispensable genes in viral genomes.
  • To enhance viral replication, spread, and immunogenicity for improved cancer therapy.

Main Methods:

  • An iterative strategy utilizing antibiotic susceptibility for genome engineering.
  • Insertional mutagenesis with the Sleeping Beauty transposon system.
  • Long-read nanopore sequencing for library generation and analysis.
  • Application to herpes simplex virus type 1 and vaccinia virus.

Main Results:

  • Successfully generated libraries of engineered oncolytic viruses.
  • Identified stable transgene insertion sites and gene deletions.
  • Demonstrated enhancement of viral safety and efficacy through genetic modification.

Conclusions:

  • The developed strategy significantly accelerates the engineering of oncolytic viruses.
  • This approach facilitates the creation of more effective and safer oncolytic virus therapies.
  • The findings pave the way for broader application of engineered viruses in cancer treatment.

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