The hyperactive Sleeping Beauty transposase SB100X improves the genetic modification of T cells to express a chimeric antigen receptor

被引:83
作者
Jin, Z. [1 ]
Maiti, S. [1 ]
Huls, H. [1 ]
Singh, H. [1 ]
Olivares, S. [1 ]
Mates, L. [2 ]
Izsvak, Z. [2 ,3 ]
Ivics, Z. [2 ,3 ]
Lee, D. A. [1 ]
Champlin, R. E.
Cooper, L. J. N. [1 ]
机构
[1] Univ Texas MD Anderson Canc Ctr, Div Pediat, Univ Texas Grad Sch Biomed Sci, Childrens Canc Hosp, Houston, TX 77030 USA
[2] Max Delbruck Ctr Mol Med, Berlin, Germany
[3] Univ Debrecen, H-4012 Debrecen, Hungary
关键词
chimeric antigen receptor; T cells; Sleeping Beauty; SB11; SB100X; CD19; SYSTEM; THERAPY; LENTIVIRUS; MALIGNANCIES; LYMPHOCYTES; SPECIFICITY; VECTORS;
D O I
10.1038/gt.2011.40
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Sleeping Beauty (SB3) transposon and transposase constitute a DNA plasmid system used for therapeutic human cell genetic engineering. Here we report a comparison of SB100X, a newly developed hyperactive SB transposase, to a previous generation SB11 transposase to achieve stable expression of a CD19-specific chimeric antigen receptor (CAR(3)) in primary human T cells. The electro-transfer of SB100X expressed from a DNA plasmid or as an introduced mRNA species had superior transposase activity in T cells based on the measurement of excision circles released after transposition and emergence of CAR expression on T cells selectively propagated upon CD19(+) artificial antigen-presenting cells. Given that T cells modified with SB100X and SB11 integrate on average one copy of the CAR transposon in each T-cell genome, the improved transposition mediated by SB100X apparently leads to an augmented founder effect of electroporated T cells with durable integration of CAR. In aggregate, SB100X improves SB transposition in primary human T cells and can be titrated with an SB transposon plasmid to improve the generation of CD19-specific CAR(+) T cells. Gene Therapy (2011) 18, 849-856; doi:10.1038/gt.2011.40; published online 31 March 2011
引用
收藏
页码:849 / 856
页数:8
相关论文
共 24 条
[21]   Sleeping Beauty Transposition From Nonintegrating Lentivirus [J].
Vink, Conrad A. ;
Gaspar, H. Bobby ;
Gabriel, Richard ;
Schmidt, Manfred ;
McIvor, R. Scott ;
Thrasher, Adrian J. ;
Qasim, Waseem .
MOLECULAR THERAPY, 2009, 17 (07) :1197-1204
[22]   RNA as a source of transposase for Sleeping Beauty-mediated gene insertion and expression in somatic cells and tissues [J].
Wilber, A ;
Frandsen, JL ;
Geurts, JL ;
Largaespada, DA ;
Hackett, PB ;
McIvor, RS .
MOLECULAR THERAPY, 2006, 13 (03) :625-630
[23]   Sleeping Beauty vector system moves toward human trials in the United States [J].
Williams, David A. .
MOLECULAR THERAPY, 2008, 16 (09) :1515-1516
[24]   Stable gene transfer and expression in cord blood-derived CD34+ hematopoietic stem and progenitor cells by a hyperactive Sleeping Beauty transposon system [J].
Xue, Xingkui ;
Huang, Xin ;
Nodland, Sonja E. ;
Mates, Lajos ;
Ma, Linan ;
Izsvak, Zsuzsanna ;
Ivics, Zoltan ;
LeBien, Tucker W. ;
McIvor, R. Scott ;
Wagner, John E. ;
Zhou, Xianzheng .
BLOOD, 2009, 114 (07) :1319-1330