Efficacy of third-party chimeric antigen receptor modified peripheral blood natural killer cells for adoptive cell therapy of B-cell precursor acute lymphoblastic leukemia

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We developed an innovative and efficient, feeder-free culture method to genetically modify and expand peripheral blood-derived NK cells with high proliferative capacity, while preserving the responsiveness of their native activating receptors. Activated peripheral blood NK cells were efficiently transduced by a retroviral vector, carrying a second-generation CAR targeting CD19. CAR expression was demonstrated across the different NK-cell subsets. CAR.CD19-NK cells display higher antileukemic activity toward CD19+ cell lines and primary blasts obtained from patients with B-cell precursor ALL compared with unmodified NK cells. In vivo animal model data showed that the antileukemia activity of CAR.CD19-NK cell is superimposable to that of CAR-T cells, with a lower xenograft toxicity profile. These data support the feasibility of generating feeder-free expanded, genetically modified peripheral blood NK cells for effective “off-the-shelf” immuno-gene-therapy, while their innate alloreactivity can be safely harnessed to potentiate allogeneic cell therapy.

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A heartfelt thanks for the helpful suggestions to Alessandro Moretta who passed away before the final acceptance. Supported by grants awarded by Fondazione AIRC per la Ricerca sul Cancro-Special Project 5 × 1000 no. 9962 (FL and LM), AIRC IG 2018 id. 21724 (FL), AIRC IG 2017 Id. 15704 (SS), AIRC IG 2017 Id. 15283 (LM), Ricerca Finalizzata GR-2013-02359212 (CQ), Ministero dell’Università e della Ricerca (grant PRIN 2017WC8499_004 to FL and SS) Ricerca Corrente (FL, CQ, and BDA).

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LM and FL share last authorship, CQ, SS, SC, and SC share first authorship of this paper. CQ, SS, AM, FL, LM, and BDA designed experimental studies, supervised the project conduction, analyzed the data, and wrote the manuscript. S.Caru., S.Carl., MG, ZA, SDC, and MS performed the in vitro experiments. S.Caru., BDA, AC, and BC performed the in vivo experiments. AP supported the patient sample processing. DO and IB cloned the retroviral vector. FM, SS, and S.Carl. performed Kir analysis and NK phenotype analysis. MA performed HLA typing of patient sample. LM and FL provided scientific advice and expertize in the immunotherapy field. GLP provided expertize in the optimization of the process for clinical translation. SG, LV, FDB, AM, and FL provided patient’s samples, medical advice, and expertize in the field of onco-hematology. SCaru., S.Carl., BDA, CQ, and SS contributed to the study design and to the analysis of experimental data.

Correspondence to B. De Angelis or F. Locatelli.

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Quintarelli, C., Sivori, S., Caruso, S. et al. Efficacy of third-party chimeric antigen receptor modified peripheral blood natural killer cells for adoptive cell therapy of B-cell precursor acute lymphoblastic leukemia. Leukemia (2019) doi:10.1038/s41375-019-0613-7

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