Hot spots of retroviral integration in human CD34+ hematopoietic cells

Cattoglio, Claudia; Facchini, Giulia; Sartori, Daniela; Antonelli, Antonella; Miccio, Annarita; Cassani, Barbara; Schmidt, Manfred; Kalle, Christof von; Howe, Steve; Thrasher, Adrian J.; Aiuti, Alessandro; Ferrari, G.; Recchia, Alessandra; Mavilio, Fulvio

doi:10.1182/blood-2007-01-068759

Cited by 244 publications

(259 citation statements)

References 33 publications

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“…It appears that the pattern of genomic integrations in cells that have been genetically modified can predict genotoxicity. HSC transduced with retroviral vectors have been infused into patients, rhesus macaques, and mice and in all cases a nonrandom pattern of vector insertions has been reported, with overrepresentation of integrations in one specific genomic locus, a complex containing the MDS1 and EVI1 genes (Cattoglio et al, 2007;Metais and Dunbar, 2008;Sellers et al, 2010). Either the MDS1-EVI1 genomic locus is a ''hot spot'' for murine leukemia virus (MLV) integration in hematopoietic stem and progenitor cells, or progenitor cells that happen to acquire an insertion in this locus have a unique propensity to engraft, persist, expand, and/or contribute to hematopoiesis after transplantation (Metais and Dunbar, 2008).…”

Section: Discussionmentioning

confidence: 99%

Infusing CD19-Directed T Cells to Augment Disease Control in Patients Undergoing Autologous Hematopoietic Stem-Cell Transplantation for Advanced B-Lymphoid Malignancies

et al. 2012

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Section: Discussionmentioning

confidence: 99%

Infusing CD19-Directed T Cells to Augment Disease Control in Patients Undergoing Autologous Hematopoietic Stem-Cell Transplantation for Advanced B-Lymphoid Malignancies

et al. 2012

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“…23,24 Briefly, genomic DNA was extracted from 1 to 5Â10 6 cells, digested with MseI and NarI to prevent amplification of internal 5¢ LTR fragments and ligated to an MseI double-strand linker. LM-PCR was performed with nested primers specific for the LTR and the linker.…”

Section: Lentiviral Insertion Analysismentioning

confidence: 99%

Risk assessment in skin gene therapy: viral–cellular fusion transcripts generated by proviral transcriptional read-through in keratinocytes transduced with self-inactivating lentiviral vectors

et al. 2011

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Cutaneous gene therapy can be envisioned through the use of keratinocyte stem cell clones in which retroviral genotoxic risks can be pre-assessed. While transactivation of cellular genes by the retroviral long terminal repeat enhancer has been proven in experimental and clinical settings, the formation of chimeric viral-cellular transcripts originated by the inefficient termination (read-through) of retroviral transcripts remains to be studied in depth. We now demonstrate the widespread presence of viral-cellular fusion transcripts derived from integrated proviruses in keratinocytes transduced with self-inactivating (SIN) retroviral vectors. We have detected high molecular weight RNAs in northern blot analysis of retroviral vector expression in individual cell clones. Characterization of some of these transcripts revealed that they originate from genes located at the proviral integration sites. One class of transcripts corresponds to fusions of the viral vectors with intronic sequences, terminating at cryptic polyadenylation sites located in introns. A second class comprises fusion transcripts with coding sequences of genes at the integration sites. These are generated through splicing from a cryptic, not previously described donor site in the lentiviral vectors to exons of cellular genes, and have the potential to encode unintended open reading frames, although they are downregulated by cellular mechanisms. Our data contribute to a better understanding of the impact of SIN lentiviral vector integration on cellular gene transcription, and will be helpful in improving the design of this type of vectors.

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“…animal as well as human models. The related problem of safety of a vector is a major hurdle (Montini et al, 2006 (Cattoglio et al, 2007) it has been proved that analysis of MLV integration patterns in natural or experimentally induced leukemias/lymphomas showed the existence of insertion sites recurrently associated with a malignant phenotype. These "common insertion sites" (CIS), also called "hotspots" which include proto-oncogenes or other genes associated with cell growth and proliferation, may present when activated a causal relationship with the establishment and/or progression of cancer.…”

Section: Introductionmentioning

confidence: 99%