Stable gene transfer and expression in cord blood-derived CD34+ hematopoietic stem and progenitor cells by a hyperactive Sleeping Beauty transposon system

Xingkui Xue, Xin Huang, Sonja E. Nodland, Lajos Mátés, Linan Ma, Zsuzsanna Izsvák, Zoltán Ivics, Tucker W. LeBien, R. Scott McIvor, John E. Wagner, Xianzheng Zhou

Research output: Contribution to journalArticlepeer-review

89 Scopus citations

Abstract

Here we report stable gene transfer in cord blood-derived CD34+ hematopoietic stem cells using a hyperactive nonviral Sleeping Beauty (SB) transposase (SB100X). In colony-forming assays, SB100X mediated the highest efficiency (24%) of stable Discosoma sp red fluorescent protein (DsRed) reporter gene transfer in committed hematopoietic progenitors compared with both the early-generation hyperactive SB11 transposase and the piggyBac transposon system (1.23% and 3.8%, respectively). In vitro differentiation assays further demonstrated that SB100X-transfected CD34+ cells can develop into DsRed+ CD4+CD8+ T (3.17%-21.84%; median, 7.97%), CD19+ B (3.83%-18.66%; median, 7.84%), CD56 +CD3- NK (3.53%-79.98%; median, 7.88%), and CD33 + myeloid (7.59%-15.63%; median, 9.48%) cells. SB100X-transfected CD34+ cells achieved approximately 46% engraftment in NOD-scid IL;cnull (NOG)mice.Twelve weeks after transplantation, 0.57% to 28.96% (median, 2.79%) and 0.49% to 34.50% (median, 5.59%) of total human CD45+ cells in the bone marrow and spleen expressed DsRed, including CD19+ B, CD14+ monocytoid, and CD33+ myeloid cell lineages. Integration site analysis revealedSBtransposon sequences in thehumanchromosomes of in vitro differentiated T, B, NK, and myeloid cells, as well as in human CD45+ cells isolated from bone marrow and spleen of transplanted NOG mice. Our results support the continuing development of SB-based gene transfer into humanhematopoietic stem cells as a modality for gene therapy.

Original languageEnglish (US)
Pages (from-to)1319-1330
Number of pages12
JournalBlood
Volume114
Issue number7
DOIs
StatePublished - 2009

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