Genotoxicity assay for gene therapy vectors in tumor prone Cdkn2a -/- mice

Eugenio Montini, Daniela Cesana

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

Integrative viral vectors are able to efficiently transduce hematopoietic stem progenitor cells allowing stable transgene expression in the entire hematopoietic system upon transplant in conditioned recipients. For these reasons, integrative vectors based on γ-retroviruses and lentiviruses have been successfully used in gene therapy clinical trials for the treatment of genetic diseases, especially blood disorders. However, in different γ-retroviral-based clinical trials, vector integration into the host cell genome triggered oncogenesis by a mechanism called insertional mutagenesis. Thus, a thorough reassessment of the safety of available gene transfer systems is a crucial outstanding issue for the whole gene therapy field. Sensitive preclinical models of vector genotoxicity are instrumental to achieve a more detailed understanding of the factors that modulate the risks of insertional mutagenesis. Here, we will describe the methodologies used to address the mutagenesis risk of vector integration using a murine in vivo genotoxicity assay based on transduction and transplantation of tumor-prone hematopoietic stem and progenitor cells.

Original languageEnglish
Pages (from-to)171-185
Number of pages15
JournalMethods in Enzymology
Volume507
DOIs
Publication statusPublished - 2012

Fingerprint

Gene therapy
Hematopoietic Stem Cells
Genetic Therapy
Tumors
Assays
Mutagenesis
Insertional Mutagenesis
Neoplasms
Clinical Trials
Hematopoietic System
Lentivirus
Inborn Genetic Diseases
Gene transfer
Retroviridae
Transgenes
Transplants
Carcinogenesis
Stem cells
Transplantation
Genome

Keywords

  • Gene deregulation by vector integration
  • Genotoxicity assays
  • Insertional mutagenesis
  • Vector integration impact

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology

Cite this

Genotoxicity assay for gene therapy vectors in tumor prone Cdkn2a -/- mice. / Montini, Eugenio; Cesana, Daniela.

In: Methods in Enzymology, Vol. 507, 2012, p. 171-185.

Research output: Contribution to journalArticle

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