Efficient lentiviral transduction method to gene modify cord blood CD8(+) T cells for cancer therapy applications.
Efficient lentiviral transduction method to gene modify cord blood CD8(+) T cells for cancer therapy applications.
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DOI：
10.1016/j.omtm.2021.03.015
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发表时间：
2021-06-11
期刊：
影响因子：
--
通讯作者：
van Til NP
中科院分区：
文献类型：
--
作者：
Lo Presti V;Cornel AM;Plantinga M;Dünnebach E;Kuball J;Boelens JJ;Nierkens S;van Til NP
利用肿瘤特异性自体T细胞的连续性T细胞疗法已经显示出用于癌症治疗的有希望的结果。然而，由于疾病进展或治疗，自体肿瘤相关抗原（TAA）特异性T细胞的数量有限和功能异常仍然是可能显著限制治疗成功的因素。使用同种异体T细胞，如脐带血（CB）来源的，克服了这些问题，但需要基因修饰，以诱导强大的和特异性的抗肿瘤作用。CB T细胞在CB库中容易获得，并且在转移时显示出低毒性、高增殖率和增加的抗白血病作用。然而，抗肿瘤基因修饰和CB T细胞的有利免疫性状的保留的组合代表了T细胞治疗产品的协调生产的主要挑战。在本文中，我们优化了用于CB CD8 + T细胞的扩增和慢病毒载体（LV）转导的方案，实现了高达83%的转导效率。在转导方案中优化LV处理的时间、培养基的选择和不同启动子的使用。LentiBOOST被确认为CB CD8 + T细胞的无毒转导增强剂，对T细胞的增殖能力和细胞活力具有轻微影响。积极的是，在肿瘤细胞识别的背景下，使用LentiBOOST不会影响细胞的功能。最后，CB CD8 + T细胞比外周血（PB）CD8 + T细胞更适合LV转导，并保持更幼稚的表型。总之，我们显示了使用LV遗传修饰CB CD8 + T细胞的有效方法，其对于用于癌症治疗的现成过继细胞治疗产品特别有用。该手稿提出了一种使用慢病毒载体和转导增强剂（LentiBOOST）对脐带血来源的CD8 + T细胞进行基因修饰的有效方法。这些发现的未来应用可以改善用于治疗癌症的同种异体T细胞疗法的产生。
Adoptive T cell therapy utilizing tumor-specific autologous T cells has shown promising results for cancer treatment. However, the limited numbers of autologous tumor-associated antigen (TAA)-specific T cells and the functional aberrancies, due to disease progression or treatment, remain factors that may significantly limit the success of the therapy. The use of allogeneic T cells, such as umbilical cord blood (CB) derived, overcomes these issues but requires gene modification to induce a robust and specific anti-tumor effect. CB T cells are readily available in CB banks and show low toxicity, high proliferation rates, and increased anti-leukemic effect upon transfer. However, the combination of anti-tumor gene modification and preservation of advantageous immunological traits of CB T cells represent major challenges for the harmonized production of T cell therapy products. In this manuscript, we optimized a protocol for expansion and lentiviral vector (LV) transduction of CB CD8+ T cells, achieving a transduction efficiency up to 83%. Timing of LV treatment, selection of culture media, and the use of different promoters were optimized in the transduction protocol. LentiBOOST was confirmed as a non-toxic transduction enhancer of CB CD8+ T cells, with minor effects on the proliferation capacity and cell viability of the T cells. Positively, the use of LentiBOOST does not affect the functionality of the cells, in the context of tumor cell recognition. Finally, CB CD8+ T cells were more amenable to LV transduction than peripheral blood (PB) CD8+ T cells and maintained a more naive phenotype. In conclusion, we show an efficient method to genetically modify CB CD8+ T cells using LV, which is especially useful for off-the-shelf adoptive cell therapy products for cancer treatment. The manuscript proposes an efficient method to gene modify cord blood-derived CD8+ T cells using lentiviral vectors and a transduction enhancer (LentiBOOST). Future application of these findings can improve the generation of allogeneic T cell therapies for the treatment of cancer.
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影响因子：
3.9
作者：
Dudley, ME;Wunderlich, JR;Rosenberg, SA
通讯作者：
Rosenberg, SA
DOI：
10.1073/pnas.0503726102
发表时间：
2005-07-05
影响因子：
11.1
作者：
Klebanoff, CA;Gattinoni, L;Restifo, NP
通讯作者：
Restifo, NP
影响因子：
8.8
作者：
Frumento, G.;Zheng, Y.;Chen, F. E.
通讯作者：
Chen, F. E.
DOI：
10.1016/j.omtm.2018.08.002
发表时间：
2018-09-21
期刊：
Molecular therapy. Methods & clinical development
影响因子：
--
作者：
Delville M;Soheili T;Bellier F;Durand A;Denis A;Lagresle-Peyrou C;Cavazzana M;Andre-Schmutz I;Six E
通讯作者：
Six E
影响因子：
6.4
作者：
Fagnoni, FF;Lozza, L;Robustelli Della Cuna, G
通讯作者：
Robustelli Della Cuna, G