The genotoxic potential of retroviral vectors is strongly modulated by vector design and integration site selection in a mouse model of HSC gene therapy
Open accessAbstract
gamma-Retroviral vectors (gammaRVs), which are commonly used in gene therapy, can trigger oncogenesis by insertional mutagenesis. Here, we have dissected the contribution of vector design and viral integration site selection (ISS) to oncogenesis using an in vivo genotoxicity assay based on transplantation of vector-transduced tumor-prone mouse hematopoietic stem/progenitor cells. By swapping genetic elements between gammaRV and lentiviral vectors (LVs), we have demonstrated that transcriptionally active long terminal repeats (LTRs) are major determinants of genotoxicity even when reconstituted in LVs and that self-inactivating (SIN) LTRs enhance the safety of gammaRVs. By comparing the genotoxicity of vectors with matched active LTRs, we were able to determine that substantially greater LV integration loads are required to approach the same oncogenic risk as gammaRVs. This difference in facilitating oncogenesis is likely to be explained by the observed preferential targeting of cancer genes by gammaRVs. This integration-site bias was intrinsic to gammaRVs, as it was also observed for SIN gammaRVs that lacked genotoxicity in our model. Our findings strongly support the use of SIN viral vector platforms and show that ISS can substantially modulate genotoxicity.
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SIN lentiviral vectors have a lower risk of insertional oncogenesis than gammaretroviral vectors in side-by-side comparisons of model systems.
“SIN lentiviral vectors were shown to have a lower risk of insertional oncogenesis than gammaretroviral vectors in side-by-side comparisons of model systems [56, 57],”
From Clinical use of lentiviral vectors (2018) · cites this paper as [56]
By swapping regions of retroviral and lentiviral LTRs, lentiviral vectors can induce insertional mutagenesis when bearing the strong promoter/enhancer elements from retroviruses.
“Montini et al. [56] demonstrated that by swapping regions of retroviral and lentiviral LTRs, lentiviral vectors can induce insertional mutagenesis when bearing the strong promoter/enhancer elements from retroviruses.”
From Clinical use of lentiviral vectors (2018) · cites this paper as [56]
Integration of proviral DNA can disrupt or enhance expression of host genes, resulting in altered and sometimes fatal outcomes.
“Equally consequential is the integration of the proviral DNA for the host, since it can either disrupt or enhance the expression of the host genes, resulting in altered and sometimes even fatal outcomes (Hacein-Bey-Abina et al. 2003, Bushman 2007, Montini et al. 2009, Desfarges and Ciuffi 2012).”
From Beyond reverse transcription: molecular mechanisms and emerging paradigms in retroviral replication (2026) · cites this paper as Montini et al. 2009
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“…consequential is the integration of the proviral DNA for the host, since it can either disrupt or enhance the expression of the host genes, resulting in altered and some-times even fatal outcomes (Hacein-Bey-Abina et al. 2003 , Bush-man 2007 , Montini et al. 2009 , Desfarges and Ciuffi 2012 ).”
- Authors
- Montini, E., Cesana, D., Schmidt, M., Sanvito, F., Bartholomae, C. C., Ranzani, M., Benedicenti, F., Sergi, L. S., Ambrosi, A., Ponzoni, M., Doglioni, C., Di Serio, C., von Kalle, C., & Naldini, L.
- Affiliations
- The San Raffaele Telethon Institute for Gene Therapy (IT)
- Vita-Salute San Raffaele University (IT)
- German Cancer Research Center (DE)
- Fondazione Telethon (IT)
- ORCID
- Venue
- Journal of Clinical Investigation
- Published
- 2009-04-01
- Volume
- 119
- Issue
- 4
- Pages
- 964-975
- ISSN
- 0021-9738 (ISSN-L)
- PMC
- PMC2662564
- OpenAlex
- W2061645642
- Cited by
- 571
- References
- 4
- Citations by year
- 2026: 12 · 2025: 16 · 2024: 16 · 2023: 17 · 2022: 24 · 2021: 28 · 2020: 31 · 2019: 25 · 2018: 17 · 2017: 30 · 2016: 30 · 2015: 29 · 2014: 33 · 2013: 58 · 2012: 55
- FWCI
- 39.19
- Type
- journal article
- Language
- en
- Open access
- green
- www.ncbi.nlm.nih.gov
- Locations
- Journal of Clinical Investigation (publishedVersion)
- PubMed Central (publishedVersion)
- PubMed (publishedVersion)
- Archivio Istituzionale della Ricerca - Università Vita-Salute San Raffaele (submittedVersion)
- Funders
- European Commission
- Fondazione Telethon
- Associazione Italiana per la Ricerca sul Cancro
- National Institutes of Health
- National Heart, Lung, and Blood Institute (P01HL053750)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Virus-based gene therapy research
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › CRISPR and Genetic Engineering
- Health Sciences › Medicine › Oncology › CAR-T cell therapy research
- Keywords
- gammaretroviral vectors, genotoxicity, insertional mutagenesis, carcinogenesis, long terminal repeat, self-inactivating vectors, lentiviral vectors, hematopoietic stem cell gene therapy, vector design
- MeSH terms
- Animals, Base Sequence, Genetic Vectors, Mice, Transgenic, Mutation, Neoplasms, Experimental, Genetic Therapy, Lentivirus, DNA Primers, Gammaretrovirus, Mice, Knockout, Hematopoietic Stem Cell Transplantation, Cyclin-Dependent Kinase Inhibitor p16, Genes, p16, Terminal Repeat Sequences, Mice
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