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Molecular evidence of lentiviral vector-mediated gene transfer into human self-renewing, multi-potent, long-term NOD/SCID repopulating hematopoietic cells.
Mol Ther. 2002 Nov; 6(5):615-26.MT

Abstract

A major challenge in gene therapy is to achieve efficient transduction of hematopoietic stem cells (HSC). It has previously been shown that lentiviral vectors (LV) transduce efficiently human cord blood-derived NOD/SCID mouse repopulating cells (SRC). Here we studied the effect of cytokines during the short ex vivo incubation with vector. Although SRC transduction was efficient without stimulation, the presence of cytokines significantly improved it. The treatment did not affect the engraftment level or the SRC frequency, but seemed to enhance SRC susceptibility to LV. SRC transduced in both conditions repopulated primary and secondary recipients, maintaining stable multi-lineage transgene expression. Using linear amplification-mediated PCR, we then analyzed vector integration in the bone marrow and CFC of the engrafted mice to monitor the clonal activity of the transduced SRC in vivo. We showed polyclonal engraftment, multi-lineage differentiation, and propagation to secondary recipients of individual SRC. We observed multiple integrations in most clones. These results provide the first formal demonstration that primitive human HSC with self-renewal and multi-lineage repopulation capacities were transduced by LV. Our findings are relevant for the design of clinical protocols that exploit this system to reach significant engraftment by genetically modified HSC in the absence of in vivo selection or strong conditioning regimens.

Authors+Show Affiliations

Laboratory for Gene Transfer and Therapy, University of Torino Medical School, Candiolo, Italy.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

12409260

Citation

Ailles, Laurie, et al. "Molecular Evidence of Lentiviral Vector-mediated Gene Transfer Into Human Self-renewing, Multi-potent, Long-term NOD/SCID Repopulating Hematopoietic Cells." Molecular Therapy : the Journal of the American Society of Gene Therapy, vol. 6, no. 5, 2002, pp. 615-26.
Ailles L, Schmidt M, Santoni de Sio FR, et al. Molecular evidence of lentiviral vector-mediated gene transfer into human self-renewing, multi-potent, long-term NOD/SCID repopulating hematopoietic cells. Mol Ther. 2002;6(5):615-26.
Ailles, L., Schmidt, M., Santoni de Sio, F. R., Glimm, H., Cavalieri, S., Bruno, S., Piacibello, W., Von Kalle, C., & Naldini, L. (2002). Molecular evidence of lentiviral vector-mediated gene transfer into human self-renewing, multi-potent, long-term NOD/SCID repopulating hematopoietic cells. Molecular Therapy : the Journal of the American Society of Gene Therapy, 6(5), 615-26.
Ailles L, et al. Molecular Evidence of Lentiviral Vector-mediated Gene Transfer Into Human Self-renewing, Multi-potent, Long-term NOD/SCID Repopulating Hematopoietic Cells. Mol Ther. 2002;6(5):615-26. PubMed PMID: 12409260.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Molecular evidence of lentiviral vector-mediated gene transfer into human self-renewing, multi-potent, long-term NOD/SCID repopulating hematopoietic cells. AU - Ailles,Laurie, AU - Schmidt,Manfred, AU - Santoni de Sio,Francesca Romana, AU - Glimm,Hanno, AU - Cavalieri,Simona, AU - Bruno,Stefania, AU - Piacibello,Wanda, AU - Von Kalle,Christof, AU - Naldini,Luigi, PY - 2002/11/1/pubmed PY - 2003/5/15/medline PY - 2002/11/1/entrez SP - 615 EP - 26 JF - Molecular therapy : the journal of the American Society of Gene Therapy JO - Mol Ther VL - 6 IS - 5 N2 - A major challenge in gene therapy is to achieve efficient transduction of hematopoietic stem cells (HSC). It has previously been shown that lentiviral vectors (LV) transduce efficiently human cord blood-derived NOD/SCID mouse repopulating cells (SRC). Here we studied the effect of cytokines during the short ex vivo incubation with vector. Although SRC transduction was efficient without stimulation, the presence of cytokines significantly improved it. The treatment did not affect the engraftment level or the SRC frequency, but seemed to enhance SRC susceptibility to LV. SRC transduced in both conditions repopulated primary and secondary recipients, maintaining stable multi-lineage transgene expression. Using linear amplification-mediated PCR, we then analyzed vector integration in the bone marrow and CFC of the engrafted mice to monitor the clonal activity of the transduced SRC in vivo. We showed polyclonal engraftment, multi-lineage differentiation, and propagation to secondary recipients of individual SRC. We observed multiple integrations in most clones. These results provide the first formal demonstration that primitive human HSC with self-renewal and multi-lineage repopulation capacities were transduced by LV. Our findings are relevant for the design of clinical protocols that exploit this system to reach significant engraftment by genetically modified HSC in the absence of in vivo selection or strong conditioning regimens. SN - 1525-0016 UR - https://www.unboundmedicine.com/medline/citation/12409260/Molecular_evidence_of_lentiviral_vector_mediated_gene_transfer_into_human_self_renewing_multi_potent_long_term_NOD/SCID_repopulating_hematopoietic_cells_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S1525001602907203 DB - PRIME DP - Unbound Medicine ER -