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Immunotherapy

TanCAR T cells targeting CD19 and CD133 efficiently eliminate MLL leukemic cells

Leukemia volume 32, pages 2012–2016 (2018)Cite this article

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Different strategies are underway to develop and optimize CARs to avoid relapses due to antigen loss [3]. One of these strategies is to use optimized CARs targeting multiple antigens in a Boolean OR-gate approach. Targeting multiple antigens allows these CARs to maintain their cytotoxic activity despite loss of one of the target cell antigens. One such approach is generation of a Tandem CAR (TanCAR), which uses two scFvs built in series in order to recognize two different targets [5]. This not only induces effective cytotoxicity when either of two antigens are presented but also synergistically enhances effector functions when both antigens are simultaneously encountered [5, 6]. This synergistic effect allows TanCAR to kill tumor cells expressing both antigens more efficiently than bystander cells expressing only one antigen. Therefore, we reasoned that TanCAR could be harnessed to not only eliminate CD19− MLL leukemic cells but also ameliorate CRS and decrease serum IL-6 levels by reducing “on-target, off-tumor” effects, ultimately preventing lineage switching. Based on this reasoning, we aimed to accomplish both goals by developing a novel TanCAR to address CD19 antigen loss in MLL-rearranged B-ALL.

To successfully apply a TanCAR strategy, we sought to identify additional markers associated with MLL leukemic cells, which maintain their expression even when a lymphoid-to-myeloid lineage switch occurs. We reasoned that CD133 could serve as a specific surface marker for MLL-rearranged B-ALL cells. CD133 is produced by the PROM1 gene, a critical target of MLL-AF4 [7, 8], and previous reports have found that it is expressed in all MLL-rearranged B-ALLs [9]. We evaluated the expression of CD133 in a panel of leukemic cell lines and found that CD133 was robustly expressed only in B-ALLs containing MLL-AF4, such as SEM and RS4;11 (Fig. 1b). CD133 was initially described as a surface antigen specific for human hematopoietic stem and progenitor cells (HSPCs) [10]. However, we found that expression of the CD133 antigen in human CD34+ HSPCs was much lower than that in B-ALL MLL leukemic cells (Fig. 1c and Supplementary Figure S1a), suggesting that a therapy window could exist if the dosage was carefully controlled in the CAR T cell therapy. Significantly, we found that expression of CD133 was maintained in CD19− MLL leukemic cells (Fig. 1d, e). These data suggested that dual targeting of CD19 and CD133 could be an effective strategy for treating and preventing CD19 antigen-loss relapses in MLL-rearranged B-ALL.

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Acknowledgements

This work was supported by the National Key Research and Development Program of China (2018YFA0107802), the National Natural Science Foundation of China (81570119 and 81370651), the Shu Guang project supported by Shanghai Municipal Education Commission and Shanghai Education Development Foundation (14SG15), the Shanghai Municipal Education Commission Gaofeng Clinical Medicine Grant (20161304), the Innovation Fund Projects of Shanghai Jiao Tong University (BXJ201809), and the Samuel Waxman Cancer Research Foundation.

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  1. These authors contributed equally: Dan Li, Yutian Hu, Zhen Jin, You Zhai

Authors and Affiliations

  1. State Key Laboratory of Medical Genomics, Shanghai Institute of Hematology, Rui Jin Hospital, School of Medicine and School of Life Sciences and Biotechnology, Shanghai Jiao Tong University, Collaborative Innovation Center of Systems Biomedicine, Collaborative Innovation Center of Hematology, Shanghai 200025, China

    Dan Li, Yutian Hu, Zhen Jin, You Zhai, Yuting Tan, Yan Sun, Shouhai Zhu, Chunjun Zhao, Bing Chen, Jiang Zhu, Zhu Chen, Saijuan Chen, Junmin Li & Han Liu

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  1. Dan Li
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Correspondence to Han Liu.

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Li, D., Hu, Y., Jin, Z. et al. TanCAR T cells targeting CD19 and CD133 efficiently eliminate MLL leukemic cells. Leukemia 32, 2012–2016 (2018). https://doi.org/10.1038/s41375-018-0212-z

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