Regulated expression of adenoviral vectors-based gene therapies: Therapeutic expression of toxins and immune-modulators

James F. Curtin, Marianela Candolfi, Mariana Puntel, Weidong Xiong, A. K.M. Muhammad, Kurt Kroeger, Sonali Mondkar, Chunyan Liu, Niyati Bondale, Pedro R. Lowenstein, Maria G. Castro

Research output: Chapter in Book/Report/Conference proceedingChapterpeer-review

Abstract

Regulatable promoter systems allow gene expression to be tightly controlled in vivo. This is highly desirable for the development of safe, efficacious adenoviral vectors that can be used to treat human diseases in the clinic. Ideally, regulatable cassettes should have minimal gene expression in the "OFF" state, and expression should quickly reach therapeutic levels in the "ON" state. In addition, the components of regulatable cassettes should be non-toxic at physiological concentrations and should not be immunogenic, especially when treating chronic illness that requires long-lasting gene expression. In this chapter, we will describe in detail protocols to develop and validate first generation (Ad) and high-capacity adenoviral (HC-Ad) vectors that express therapeutic genes under the control of the TetON regulatable system. Our laboratory has successfully used these protocols to regulate the expression of marker genes, immune stimulatory genes, and toxins for cancer gene therapeutics, i.e., glioma that is a deadly form of brain cancer. We have shown that this third generation TetON regulatable system, incorporating a doxycycline (DOX)-sensitive inducer and silencer, is non-toxic, relatively non-immunogenic, and can tightly regulate reporter transgene expression downstream of a TRE promoter from adenoviral vectors in vitro and also in vivo.

Original languageEnglish
Title of host publicationGene Therapy Protocols
Subtitle of host publicationDesign and Characterization of Gene Transfer Vectors
PublisherHumana Press
Pages239-266
Number of pages28
ISBN (Print)9781603272476
DOIs
Publication statusPublished - 2008
Externally publishedYes

Publication series

NameMethods in Molecular Biology
Volume434
ISSN (Print)1064-3745

Keywords

  • Adenoviral vectors
  • Doxycycline
  • Gene therapy
  • Glioma
  • Inducible
  • Pseudomonas exotoxin A (PE)
  • Regulatable
  • TetON
  • Tetracycline

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