Redirecting specificity of T-cell Populations for CD19 using the Sleeping beauty system
Redirecting specificity of T-cell Populations for CD19 using the Sleeping beauty system
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DOI：
10.1158/0008-5472.can-07-5600
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发表时间：
2008-04-15
期刊：
影响因子：
11.2
通讯作者：
Cooper, Laurence J. N.
中科院分区：
文献类型：
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作者：
Singh, Harjeet;Manuri, Pallavi R.;Cooper, Laurence J. N.
对临床级T细胞进行遗传修饰以增强功能，包括重定向对所需抗原的特异性。我们和其他人已经引入了嵌合抗原受体（CAR），使T细胞能够识别谱系特异性肿瘤抗原，如CD 19，目前正在进行早期人体试验，以评估其安全性和可行性。然而，下一代临床研究的一个重要障碍是开发能够遗传修饰广泛T细胞群体的合适CAR表达载体。T细胞的转导相对有效，但它需要专门制造昂贵的临床级重组病毒。裸DNA质粒的电转移提供了一种具有成本效益的替代方法，但转基因整合的效率低下要求在杀细胞浓度的药物下进行离体选择，以强制选择基因的表达，从而获得临床上有意义数量的CAR（+）T细胞。我们报告了一种有效产生具有重定向特异性的T细胞的新方法，从睡美人转座子/转座酶系统引入DNA质粒，在记忆和效应T细胞中直接表达CD 19特异性CAR，而无需药物选择。当与CD 19（+）人工抗原呈递细胞上的数值扩增相结合时，这种基因转移方法导致表达CAR的CD 4（+）和CD 8（+）T细胞快速生长，以重定向对CD 19（+）肿瘤细胞的特异性。
Genetic modification of clinical-grade T cells is undertaken to augment function, including redirecting specificity for desired antigen. We and others have introduced a chimeric antigen receptor (CAR) to enable T cells to recognize lineage-specific tumor antigen, such as CD19, and early-phase human trials are currently assessing safety and feasibility. However, a Significant barrier to next-generation clinical studies is developing a suitable CAR expression vector capable of genetically modifying a broad population of T cells. Transduction of T cells is relatively efficient but it requires specialized manufacture of expensive clinical grade recombinant virus. Electrotransfer of naked DNA plasmid offers a cost-effective alternative approach, but the inefficiency of transgene integration mandates ex vivo selection under cytocidal concentrations of drug to enforce expression of selection genes to achieve clinically meaningful numbers of CAR(+) T cells. We report a new approach to efficiently generating T cells with redirected specificity, introducing DNA plasmids from the Sleeping Beauty transposon/transposase system to directly express a CD19-specific CAR in memory and effector T cells without drug selection. When coupled with numerical expansion on CD19(+) artificial antigen-presenting cells, this gene transfer method results in rapid outgrowth of CD4(+) and CD8(+) T cells expressing CAR to redirect specificity for CD19(+) tumor cells.