NK cells; complement-dependent cytotoxicity; immunotherapy; phagocytosis; xenograft model; Immunology and Allergy; Immunology
Abstract :
[en] [en] PURPOSE: Directing selective complement activation towards tumor cells is an attractive strategy to promote their elimination. We have generated complement-activating multimeric immunotherapeutic complexes (CoMiX), stimulating either the alternative pathway (via Factor H Related protein 4 (FHR4)) or the classical pathway (via triple Fc dimers) on HER2-expressing tumor cells.
METHODS: We used the C-terminal α-chain multimerizing scaffold of the C4 binding protein (C4bp) to generate CoMiX-FHR4 as well as CoMiX-Fc with 2 different anti-HER2 VHH, VHH(T) and VHH(P), recognizing trastuzumab- or pertuzumab-competing epitopes, respectively. The different CoMiX were compared in vitro for C3b and C5b9 depositions, complement-dependent cytotoxicity (CDC), and their ability to activate NK cells and phagocytosis by macrophages. We further explored their therapeutic efficacy on human BT474 tumor xenografts established in nude mice.
RESULTS: CoMiX-FHR4/VHH(T) and -FHR4/VHH(P) lead to the highest C3b and C5b9 depositions and CDC on BT474 tumor cells (p<0.0001), both individually and in combinations with their CoMiX-Fc counterparts, surpassing the low complement activating capacity of trastuzumab and pertuzumab. All CoMiX induced BT474 cell death and phagocytosis of tumor cells by macrophages while CoMiX-Fc also stimulated NK cell activation. In human BT474 xenografts sensitive to trastuzumab, CoMiX induced a massive C3b deposition 6 hours after injection. CoMiX-FHR4 reduced the tumor volume compared to controls (p< 0.05) but to a lesser extent than trastuzumab (p< 0.001) while CoMiX-VHH(P)/Fc led to a tumor volume reduction similar to pertuzumab. Combinations of two CoMiX-FHR4 or two CoMiX-Fc were more potent, similarly to the combination of trastuzumab and pertuzumab, leading to increased NK cell infiltration in xenografts. Importantly, CoMiX-FHR4 was still active against trastuzumab-resistant xenografts, delaying tumor growth and inducing a large NK cell infiltration.
CONCLUSION: We showed here that directed complement activation on tumor cells is an alternative to therapeutic antibodies for future combination therapies upon resistance to standard-of-care treatment.
Disciplines :
Oncology
Author, co-author :
Seguin-Devaux, Carole ; Department of Infection and Immunity, Luxembourg Institute of Health, Esch sur Alzette, Luxembourg
BRANDUS, Bianca ; University of Luxembourg ; Department of Infection and Immunity, Luxembourg Institute of Health, Esch sur Alzette, Luxembourg
PLESSERIA, Jean-Marc ; University of Luxembourg ; Department of Infection and Immunity, Luxembourg Institute of Health, Esch sur Alzette, Luxembourg
Iserentant, Gilles; Department of Infection and Immunity, Luxembourg Institute of Health, Esch sur Alzette, Luxembourg
Servais, Jean-Yves; Department of Infection and Immunity, Luxembourg Institute of Health, Esch sur Alzette, Luxembourg
Pitiot, Aubin; Department of Infection and Immunity, Luxembourg Institute of Health, Esch sur Alzette, Luxembourg
Kanli, Georgia; Department of Oncology, Luxembourg Institute of Health, Luxembourg, Luxembourg
BEHRMANN, Iris ; University of Luxembourg > Faculty of Science, Technology and Medicine (FSTM) > Department of Life Sciences and Medicine (DLSM)
Schober, Rafaëla; Department of Infection and Immunity, Luxembourg Institute of Health, Esch sur Alzette, Luxembourg
ZIMMER, Jacques ; University of Luxembourg ; Department of Infection and Immunity, Luxembourg Institute of Health, Esch sur Alzette, Luxembourg
Cohen, Jacques H M; LRN EA4682, University of Reims Champagne Ardennes, Reims, France
DERVILLEZ, Xavier ; University of Luxembourg > Faculty of Science, Technology and Medicine > Department of Life Sciences and Medicine ; Department of Infection and Immunity, Luxembourg Institute of Health, Esch sur Alzette, Luxembourg
External co-authors :
yes
Language :
English
Title :
Directed-Complement Activation as a Novel Immunotherapeutic Approach for HER2-Breast Cancer.
The study was supported by the \u201CFonds National de la Recherche\u201D (POC17/12252709/COMIX and PRIDE17/11823097/MICROH-DTU for the funding of Bianca Brandus) and the Ministry of Higher Education and Research of Luxembourg(LIH GBB 98000005). We thank the participants who has given their PBMCs for research to the Luxembourg Red Cross and the Luxembourg Red Cross for the collection of the samples. We would like to acknowledge Brigitte Reveil for her technical assistance and Pr B\u00E9atrice Clemenceau, University of Nantes, for providing the NK92humCD16 cell line.
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