1. Strategy to prevent epitope masking in CAR.CD19+ B-cell leukemia blasts
- Author
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Pietro Merli, Gianni Cazzaniga, Franco Locatelli, Vittorio Nunes, Zeinab Abbaszadeh, Alberto Orfao, Matilde Sinibaldi, Mattia Algeri, Frederikke Isa Marin, Maria Herrero-Garcia, Stefano Di Cecca, Paolo Marcatili, Biagio De Angelis, Concetta Quintarelli, Sara Gutiérrez-Herrero, Luciana Vinti, Lourdes Martín-Martín, Roselia Ciccone, Biancamaria Cembrola, Simona Songia, Simona Caruso, Iolanda Boffa, Simona Manni, Valentina Bertaina, Marika Guercio, Antonio Camera, Francesca Del Bufalo, Marco Ruella, Quintarelli, C, Guercio, M, Manni, S, Boffa, I, Sinibaldi, M, DI Cecca, S, Caruso, S, Abbaszadeh, Z, Camera, A, Cembrola, B, Ciccone, R, Orfao, A, Martin-Martin, L, Gutierrez-Herrero, S, Herrero-Garcia, M, Cazzaniga, G, Nunes, V, Songia, S, Marcatili, P, Marin, F, Ruella, M, Bertaina, V, Vinti, L, Del Bufalo, F, Algeri, M, Merli, P, De Angelis, B, Locatelli, F, Cancer Research UK, Associazione Italiana per la Ricerca sul Cancro, Asociación Española Contra el Cáncer, and Ministero della Salute
- Subjects
0301 basic medicine ,Cancer Research ,receptor ,receptors ,Epitope ,Epitopes ,Mice ,0302 clinical medicine ,hemic and lymphatic diseases ,Immunology and Allergy ,hematologic neoplasms ,RC254-282 ,Receptors, Chimeric Antigen ,biology ,medicine.diagnostic_test ,Chemistry ,Neoplasms. Tumors. Oncology. Including cancer and carcinogens ,Leukemia ,Oncology ,Settore MED/38 - PEDIATRIA GENERALE E SPECIALISTICA ,030220 oncology & carcinogenesis ,translational medical research ,Molecular Medicine ,immunotherapy ,Immunology ,cell engineering ,adoptive ,CD19 ,Flow cytometry ,03 medical and health sciences ,In vivo ,Cell Line, Tumor ,Leukemia, B-Cell ,medicine ,Animals ,Humans ,Pharmacology ,Immune Cell Therapies and Immune Cell Engineering ,medicine.disease ,Chimeric antigen receptor ,In vitro ,Disease Models, Animal ,030104 developmental biology ,B-cell leukemia ,chimeric antigen ,Cancer research ,biology.protein ,human activities ,hematologic neoplasm - Abstract
[Abstract]: Chimeric antigen receptor T-cells (CAR T-cells) for the treatment of relapsing/refractory B-cell precursor acute lymphoblastic leukemia have led to exciting clinical results. However, CAR T-cell approaches revealed a potential risk of CD19-/CAR+ leukemic relapse due to inadvertent transduction of leukemia cells. [Methods]: We evaluated the impact of a high percentage of leukemia blast contamination in patient-derived starting material (SM) on CAR T-cell drug product (DP) manufacturing. In vitro as well as in vivo models were employed to identify characteristics of the construct associated with better profile of safety in case of inadvertent B-cell leukemia transduction during CAR T-cell manufacturing. [Results]: The presence of large amounts of CD19+ cells in SM did not affect the transduction level of DPs, as well as the CAR T-cell rate of expansion at the end of standard production of 14 days. DPs were deeply characterized by flow cytometry and molecular biology for Ig-rearrangements, showing that the level of B-cell contamination in DPs did not correlate with the percentage of CD19+ cells in SM, in the studied patient cohort. Moreover, we investigated whether CAR design may affect the control of CAR+ leukemia cells. We provided evidences that CAR.CD19 short linker (SL) prevents complete epitope masking in CD19+CAR+ leukemia cells and we demonstrated in vitro and in vivo that CD19 +CAR(SL)+leukemic cells are killed by CAR.CD19 T-cells. [Conclusions]: Taken together, these data suggest that a VL-VH SL may result in a safe CAR-T product, even when manufacturing starts from biological materials characterized by heavy contamination of leukemia blasts., The experimental work was supported by grants awarded by Ricerca Finalizzata GR-2013 02359212 (CQ), GR-2016-02364546 (BDA), RF-2016- 02364388 (FL), Accelerator Award-Cancer Research UK/AIRC/AECC-INCAR project (FL and AO), Associazione Italiana Ricerca per la Ricerca sul Cancro (AIRC)-Special Project 5×1000 no. 9962 (FL), AIRC IG 2018 id. 21724 (FL), Ricerca Corrente (FL, CQ, BDA), Ministero dell’Università e della Ricerca (Grant PRIN 2017 to FL); Italian Healthy Ministry project on CAR T RCR-2019-23669115 (coordinator FL), Independent Research grant AIFA (FL PI: 2016 call).
- Published
- 2021