ELIMINATION OF EGFR-OVEREXPRESSING CANCER CELLS BY CD32 CHIMERIC RECEPTOR T CELLS IN COMBINATION WITH CETUXIMAB OR PANITUMUMAB

Caratelli, S.; Arriga, R.; Sconocchia, T.; Ottaviani, A.; Lanzilli, G.; Pastore, D.; Cenciarelli, C.; Venditti, A.; Del Principe, M. I.; Lauro, D.; Landoni, E.; Du, H.; Savoldo, B.; Ferrone, S.; Dotti, G.; Sconocchia, G.

doi:10.1101/581017

bioRxiv · 10.1101/581017

ELIMINATION OF EGFR-OVEREXPRESSING CANCER CELLS BY CD32 CHIMERIC RECEPTOR T CELLS IN COMBINATION WITH CETUXIMAB OR PANITUMUMAB

Caratelli, S.·Arriga, R.·Sconocchia, T.·Ottaviani, A.·Lanzilli, G.·Pastore, D.·Cenciarelli, C.·Venditti, A.·Del Principe, M. I.·Lauro, D.·Landoni, E.·Du, H.·Savoldo, B.·Ferrone, S.·Dotti, G.·Sconocchia, G.

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Abstract

Cetuximab and panitumumab bind the human epidermal growth factor receptor (EGFR). While the chimeric cetuximab (IgG1) triggers antibody-dependent-cellular-cytotoxicity (ADCC) of EGFR positive target cells, panitumumab (a human IgG2) does not. The inability of panitumumab to trigger ADCC reflects a poor binding affinity of human IgG2 Fc for the Fc{gamma}RIII (CD16) on NK cells. However, both human IgG1 and IgG2 bind the Fc{gamma}RII (CD32) to a similar extent. Here, we have compared the ability of T cells, engineered with a novel low-affinity CD32131R -chimeric receptor (CR), and those engineered with the low-affinity CD16158F-CR T cells in eliminating EGFR positive epithelial cancer cells (ECCs) in combination with cetuximab or panitumumab. Following T cell transduction, the percentage of CD32131R-CR T cells was (74{+/-}10) significantly higher than that of CD16158F-CR T cells (46{+/-}15). Only CD32131R-CR T cells bound panitumumab. CD32131R-CR T cells combined with the mAb 8.26 (anti-CD32) and CD16158F-CR T cells combined with the mAb 3g8 (anti-CD16) eliminated colorectal carcinoma (CRC), HCT116Fc{gamma}R+ cells, in a reverse ADCC assay in vitro. Cross-linking of CD32131R-CR on T cells by cetuximab or panitumumab and CD16158F-CR T cells by cetuximab induced elimination of triple negative breast cancer (TNBC) MDA-MB-468 cells, and secretion of IFN gamma (IFN{gamma}) and tumor necrosis factor alpha (TNF). Neither cetuximab nor panitumumab induced Fc{gamma}-CR T anti-tumor activity against KRAS-mutated HCT116, non-small-cell-lung-cancer, A549 and TNBC, MDA-MB-231 cells. ADCC of Fc{gamma}-CR T cells was significantly associated with the over-expression of EGFR on ECCs. In conclusion, CD32131R-CR T cells are efficiently redirected by cetuximab or panitumumab against BC cells overexpressing EGFR.\n\nArticle categoryTumor Immunology and Microenvironment\n\nNovelty and ImpactMonoclonal antibody-redirected Fc{gamma}-CR T cell immunotherapy represents a promising approach in the fight against cancer. Here, we expand the application of this methodology to TNBC overexpressing the EGFR utilizing a novel CD32A131R-CR in combination with anti-EGFR mAbs. Our study supports the use of CD32A131R-CR T cells combined with panitumumab or cetuximab for targeting TNBC cells overexpressing the EGFR. Our results may be utilized as a platform for the rational design of therapies targeting TNBC overexpressing EGFR.

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Caratelli, S., Arriga, R., Sconocchia, T., Ottaviani, A., Lanzilli, G., Pastore, D., Cenciarelli, C., Venditti, A., Del Principe, M. I., Lauro, D., Landoni, E., Du, H., Savoldo, B., Ferrone, S., Dotti, G., Sconocchia, G.. 2019-03-18. ELIMINATION OF EGFR-OVEREXPRESSING CANCER CELLS BY CD32 CHIMERIC RECEPTOR T CELLS IN COMBINATION WITH CETUXIMAB OR PANITUMUMAB. https://doi.org/10.1101/581017

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