Chimeric antigen receptor–induced BCL11B suppression propagates NK-like cell development
Marcel Maluski, … , Marcel R.M. van den Brink, Martin G. Sauer
Marcel Maluski, … , Marcel R.M. van den Brink, Martin G. Sauer
Published September 3, 2019
Citation Information: J Clin Invest. 2019;129(12):5108-5122. https://doi.org/10.1172/JCI126350.
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Research Article Immunology

Chimeric antigen receptor–induced BCL11B suppression propagates NK-like cell development

Abstract

The transcription factor B cell CLL/lymphoma 11B (BCL11B) is indispensable for T lineage development of lymphoid progenitors. Here, we show that chimeric antigen receptor (CAR) expression during early phases of ex vivo generation of lymphoid progenitors suppressed BCL11B, leading to suppression of T cell–associated gene expression and acquisition of NK cell–like properties. Upon adoptive transfer into hematopoietic stem cell transplant recipients, CAR-expressing lymphoid progenitors differentiated into CAR-induced killer (CARiK) cells that mediated potent antigen-directed antileukemic activity even across MHC barriers. CD28 and active immunoreceptor tyrosine–based activation motifs were critical for a functional CARiK phenotype. These results give important insights into differentiation of murine and human lymphoid progenitors driven by synthetic CAR transgene expression and encourage further evaluation of ex vivo–generated CARiK cells for targeted immunotherapy.

Authors

Marcel Maluski, Arnab Ghosh, Jessica Herbst, Vanessa Scholl, Rolf Baumann, Jochen Huehn, Robert Geffers, Johann Meyer, Holger Maul, Britta Eiz-Vesper, Andreas Krueger, Axel Schambach, Marcel R.M. van den Brink, Martin G. Sauer

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Figure 6

CARs containing CD28 costimulatory domain induce killer cells with superior functionality.

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CARs containing CD28 costimulatory domain induce killer cells with super...
(A) Design of the im19delta, im19z1, im19z3, and im1928z3 constructs. All CAR constructs were linked to an IRES dTomato cassette. (B and C) Representative FACS plots (B) and respective CD25midCD44+, DN2 (CD25+CD44+), and DN3 (CD25+CD44–) populations (C) of lymphoid progenitors engineered with the indicated CAR construct (color coded as indicated) on day 20 of in vitro culture. (D) Frequencies of CD122+NK1.1+ CARiK cells on day 20 of in vitro culture. Tom+ cells were analyzed. (B–D) Data from 1 of 2 independent experiments measured in triplicates are shown. (E–G) Irradiated B6 recipients were reconstituted with 3 × 106 B6 TCD-BM and cotransplanted with 8 × 106 lymphoid progenitors that had been engineered with the indicated CAR constructs. (E) BM cells were analyzed for numbers of CD122+ (left) and NK1.1+NKp46+ cells (right) within on day 14. im19delta, im19z3, and im1928z3 (n = 5 mice); im19z1 (n = 6); im1928z1 (n = 4). (F) CD19+ B cells were quantified in BM (left) and spleens (right) on day 28. n = 5 mice for each group. (G) CD19+ B cells in the peripheral blood were determined in 7- to 14-day intervals. Analysis at each time point was done on n = 4–5 mice per group. (C–F) Analysis was done using 1-way ANOVA with Tukey’s post test. Data represent mean ± SEM. (H) Irradiated B6 recipients were transplanted with 3 × 106 B6 TCD-BM only (n = 10) or additionally with 8 × 106 CAR-expressing lymphoid progenitors (n = 10). Mice were challenged with 1.2 × 106 C1498-mCD19 cells on day 21 after transplantation and monitored for survival. Survival curves were compared using Mantel-Cox (log-rank) test. *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001.