Nature Biotechnology 24, 687 - 696 (2006)
Published online: 28 May 2006; | doi:10.1038/nbt1216
Hematopoietic stem cell gene transfer in a tumor-prone mouse model uncovers low genotoxicity of lentiviral vector integrationEugenio Montini1, Daniela Cesana1, 2, Manfred Schmidt3, 4, Francesca Sanvito5, Maurilio Ponzoni5, Cynthia Bartholomae4, Lucia Sergi Sergi1, Fabrizio Benedicenti1, Alessandro Ambrosi6, Clelia Di Serio6, Claudio Doglioni2, 5, Christof von Kalle3, 4, 7
& Luigi Naldini1, 21
San Raffaele-Telethon Institute for Gene Therapy, via Olgettina 58, 20132, Milan, Italy. 2
Vita Salute San Raffaele University, via Olgettina 58, 20132, Milan, Italy. 3
Department of Internal Medicine I, University Hospital, Hugstetterstrasse 55, 79106 Freiburg, Germany. 4
National Center for Tumor Diseases (NCT), im Neuenheimer Feld 350, 69120 Heidelberg, Germany. 5
Department of Pathology, San Raffaele Hospital, via Olgettina 60, 20132, Milan, Italy. 6
University Centre of Statistics for Biomedical Sciences, Vita Salute San Raffaele University, via Olgettina 58, 20132, Milan, Italy. 7
Cincinnati Children's Research Foundation, Molecular and Gene Therapy Program, 3333 Burnet Avenue, Cincinnati, Ohio 45229, USA.
Correspondence should be addressed to Luigi Naldini naldini.luigi@hsr.it Insertional mutagenesis represents a major hurdle to gene therapy and necessitates sensitive preclinical genotoxicity assays. Cdkn2a
-/- mice are susceptible to a broad range of cancer-triggering genetic lesions. We exploited hematopoietic stem cells from these tumor-prone mice to assess the oncogenicity of prototypical retroviral and lentiviral vectors. We transduced hematopoietic stem cells in matched clinically relevant conditions, and compared integration site selection and tumor development in transplanted mice. Retroviral vectors triggered dose-dependent acceleration of tumor onset contingent on long terminal repeat activity. Insertions at oncogenes and cell-cycle genes were enriched in early-onset tumors, indicating cooperation in tumorigenesis. In contrast, tumorigenesis was unaffected by lentiviral vectors and did not enrich for specific integrants, despite the higher integration load and robust expression of lentiviral vectors in all hematopoietic lineages. Our results validate a much-needed platform to assess vector safety and provide direct evidence that prototypical lentiviral vectors have low oncogenic potential, highlighting a major rationale for application to gene therapy.
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