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CD28/4-1BB CD123 CAR T cells in blastic plasmacytoid dendritic cell neoplasm


Blastic plasmacytoid dendritic cell neoplasm (BPDCN) is associated with a remarkably poor prognosis and with no treatment consensus. The identification of relevant therapeutic targets is challenging. Here, we investigated the immune functions, antileukemia efficacy and safety of CD28/4-1BB CAR T cells targeting CD123 the interleukin (IL)-3 receptor alpha chain which is overexpressed on BPDCN. We demonstrated that both retroviral and lentiviral engineering CD28/4-1BB CD123 CAR T cells exhibit effector functions against BPDCN cells through CD123 antigen recognition and that they efficiently kill BPDCN cell lines and BPDCN-derived PDX cells. In vivo, CD28/4-1BB CD123 CAR T-cell therapy displayed strong efficacy by promoting a decrease of BPDCN blast burden. Furthermore we showed that T cells from BPDCN patient transduced with CD28/4-1BB CD123 CAR successfully eliminate autologous BPDCN blasts in vitro. Finally, we demonstrated in humanized mouse models that these effector CAR T cells exert low or no cytotoxicity against various subsets of normal cells with low CD123 expression, indicating a potentially low on-target/off-tumor toxicity effect. Collectively, our data support the further evaluation for clinical assessment of CD28/4-1BB CD123 CAR T cells in BPDCN neoplasm.

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Fig. 1: Design and production of CD123 CAR T cells.
Fig. 2: CD123R CAR T cells induced specific cytotoxicity against BPDCN cell lines and BPDCN patient-derived xenograft cells.
Fig. 3: Functional characterization of CD123R CAR T cells in coculture with CD123+ target cells.
Fig. 4: In vivo antitumor activity of CD123L CAR T cells against CAL-1 and PDX.
Fig. 5: Evaluation of CD123 expression in normal cells compared with BPDCN cells.
Fig. 6: Evaluation of toxicity of CD123L CAR T cells against cells expressing CD123 at a low level.
Fig. 7: CD123L CAR T cells from a patient with BPDCN and induction of specific cytotoxicity against autologous BPDCN blasts.


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The authors would like to thank the animal technicians for their expertise in animal care and management. We thank Franck Monnien and the Biobank BB-0033-00024 “Tumorothèque Régionale de Franche-Comté (TRFC)” for providing tissue samples, Diaclone for collaboration for antibodies production, and Stemline for SL-401 providing. This study was supported by Programme de Recherche Translationnelle INCa (PRTK N°PRT-K-15-175), by the MiMedI project funded by BPI France (Grant No. DOS0060162/00) and the European Union through the European Regional Development Fund of the Region Bourgogne-Franche-Comte (Grant No. FC0013440), by the Region Bourgogne-Franche-Comté (IVIS Lumina Series III bioimager to P.S. #2016-0196), by the National Cancer Institute (INCa) DM/FC/sl/RT07 grant to CR and the Association Laurette Fugain (12/09) to EM.

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EBR, MF, SB, JMC, FA, FT, MP, and AJ performed the experiments; EBR, JMC, MF, and SB analyzed data; EBR and MF performed statistical analysis; FB, JRP, and KYHWY assisted with in vivo experiments; FR, TP, CM, and SVD provided help with immunohistochemistry and transcriptomic analysis. TP, CR, EM, FF, EB, and MM provided patient samples and expertise. FR, CM, SVD, YG, PS, WW, MD, PLD, ML, JC, DB, MP, ED, ED, PS, CF, and FAD provided guidance and expertise in their respective areas of study. EBR, MF, and FGO wrote the manuscript, and FGO, OA, ED, PS, and FAD commented on the manuscript. FGO, FAD, and OA supervised the research. All authors provided input and edited and approved the final version of the manuscript.

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Correspondence to Francine Garnache-Ottou.

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Bôle-Richard, E., Fredon, M., Biichlé, S. et al. CD28/4-1BB CD123 CAR T cells in blastic plasmacytoid dendritic cell neoplasm. Leukemia 34, 3228–3241 (2020).

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