An effective platform for cancer immunotherapy: pooled knockin targeting for genome engineering
An effective platform for cancer immunotherapy: pooled knockin targeting for genome engineering
复制标题
癌症免疫治疗的有效平台：基因组工程的混合敲入靶向
DOI：
10.1038/s41392-020-0208-9
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发表时间：
2020-06
影响因子：
39.3
通讯作者：
Zhou F
中科院分区：
文献类型：
--
作者：
Jin K;Zhang L;Zhou F
最近，西奥多·L·罗斯（Theodore L. Roth）等人在《细胞》（Cell）杂志上发表的一篇论文报道了一个平台的开发，该平台用于评估针对基因组中特定基因座的混合敲入构建体的功能效应。它还提供了一种有助于发现（的策略）
注：原文最后一个单词“discovery”是名词形式，翻译时补充了括号里的内容使语义更通顺完整。
Recently, a paper published in Cell by Theodore L. Roth et al. reported the development of a platform to assess the functional effects of pooled knockin constructs targeting a specific locus in the genome. It also provided a strategy that allows discovery of novel synthetic constructs in the pools to enable engineering T cells with gain of function and promote antitumor activity of T cells in vivo. 1 The immune related components in the tumor microenvironment (TME) are complex. Among them, tumor-infiltrating lymphocytes (TILs) are recognized as one of the key pillars of cancer immunotherapy. Infiltrating T cells, including CD4+ and CD8+ subsets, participate in coordination in shaping tumor immunity and influencing the fate of cancer. Therefore, developing therapeutic approaches targeting T cells that ultimately enhance the antitumor activity of TILs appears to be particularly important. Immune checkpoint therapy (ICT) performed by blocking CTLA-4, PD-1, or PD-L1 has been demonstrated as an effective strategy for clinical cancer treatment. However, the main obstacle for limiting the effectiveness of ICT in the clinical setting is T cell exhaustion. Tumor cells dictate immunosuppressive components (immune cells such as Treg cells, M2 macrophages, and myeloid-derived suppressor cells (MDSCs) as well as cytokines including TGF-β and IL-10) to orchestrate an immunosuppressive TME, which restrains entry and activity of the effector T cells and hinders ICT (Fig. 1). Particularly, blocking TGF-β in the osseous TME restores antitumor activity of ICT in a bone prostate cancer model. 2 In addition, cellular therapeutics utilizing CD8+ T cells with chimeric antigen receptors (CARs) have also exhibited clinical success against hematopoietic malignancies. Nevertheless, CAR-T therapy still has some disadvantages. First, searching for unique neoantigens in tumors is difficult and time consuming because of tumor heterogeneity accompanied with immunoediting during cancer evolution. 3 Second, CAR-T has not proven effective in the treatment of solid tumors. Third, the manufacturing process for CAR-T is complex and expensive. Hence, developing new methods for constructing genome-engineered human T cells holds great potential for the next generation of cellbased immunotherapies for cancer treatment. In 2015, Alexander Marsona group in collaboration with Jennifer Doudna developed a robust CRISPR/Cas9-based technology that enabled both “knockout” and “knockin” genome editing in primary human T cells. 4 The core elements for this system are Cas9 ribonucleoproteins (RNP), a complex of recombinant Cas9 proteins and an in vitro transcribed sgRNA. For “knockin” editing, a homology-directed repair template (HDRT) is needed. There are several characteristics for this system (Fig. 1). First, this CRISPR/Cas9 genome-targeting technology utilizes electroporation instead of recombinant viral vectors, which allows rapid and efficient insertion of DNA sequences ranging from 2 kb to 3 kb without notable cell toxicity. 1, 5 Moreover, this non-viral genome-targeting method allows the correction of point mutations in the original genome. 4 Second, during the “knockin” procedure, DNA cassettes can be introduced to specific genomic sites remolding the function of T cells. In the current study, it is by using this system that Alexander Marsona group succeeds in integrating a pool of functional DNA cassettes into the first exon of the T cell receptor (TCR)-α constant region (TRAC). Thereafter, the endogenous TCR locus is replaced with a new TCR that redirects the T cells to recognize a specific cancer antigen. Third, multiple pooled knockin templates with specific barcodes …
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影响因子：
64.5
作者：
Roth, Theodore L.;Li, P. Jonathan;Marson, Alexander
通讯作者：
Marson, Alexander
DOI：
10.1073/pnas.1512503112
发表时间：
2015-08-18
影响因子：
11.1
作者：
Schumann, Kathrin;Lin, Steven;Marson, Alexander
通讯作者：
Marson, Alexander
影响因子：
64.8
作者：
Roth TL;Puig-Saus C;Yu R;Shifrut E;Carnevale J;Li PJ;Hiatt J;Saco J;Krystofinski P;Li H;Tobin V;Nguyen DN;Lee MR;Putnam AL;Ferris AL;Chen JW;Schickel JN;Pellerin L;Carmody D;Alkorta-Aranburu G;Del Gaudio D;Matsumoto H;Morell M;Mao Y;Cho M;Quadros RM;Gurumurthy CB;Smith B;Haugwitz M;Hughes SH;Weissman JS;Schumann K;Esensten JH;May AP;Ashworth A;Kupfer GM;Greeley SAW;Bacchetta R;Meffre E;Roncarolo MG;Romberg N;Herold KC;Ribas A;Leonetti MD;Marson A
通讯作者：
Marson A
影响因子：
64.5
作者：
Jiao, Shiping;Subudhi, Sumit K.;Sharma, Padmanee
通讯作者：
Sharma, Padmanee
影响因子：
64.5
作者：
Angelova, Mihaela;Mlecnik, Bernhard;Galon, Jerome
通讯作者：
Galon, Jerome