Crizotinib-induced immunogenic cell death in non-small cell lung cancer

Abstract

Immunogenic cell death (ICD) converts dying cancer cells into a therapeutic vaccine and stimulates antitumor immune responses. Here we unravel the results of an unbiased screen identifying high-dose (10 µM) crizotinib as an ICD-inducing tyrosine kinase inhibitor that has exceptional antineoplastic activity when combined with non-ICD inducing chemotherapeutics like cisplatin. The combination of cisplatin and high-dose crizotinib induces ICD in non-small cell lung carcinoma (NSCLC) cells and effectively controls the growth of distinct (transplantable, carcinogen- or oncogene induced) orthotopic NSCLC models. These anticancer effects are linked to increased T lymphocyte infiltration and are abolished by T cell depletion or interferon-γ neutralization. Crizotinib plus cisplatin leads to an increase in the expression of PD-1 and PD-L1 in tumors, coupled to a strong sensitization of NSCLC to immunotherapy with PD-1 antibodies. Hence, a sequential combination treatment consisting in conventional chemotherapy together with crizotinib, followed by immune checkpoint blockade may be active against NSCLC.

Platinum Derivatives Effects on Anticancer Immune Response

“…Although this tyrosine kinase inhibitor is able by itself to induce CRT exposure, ATP and HMGB1 release by NSCLCs, a vaccination effect of this compound requires cisplatin (or mitomycin C) treatment. It is worth noting that the combination of crizotinib and cisplatin together with anti-PD-1 and anti-CTLA4 immunotherapy has an important vaccine and antitumor effect [90]. Targeting another kinase, the serine/threonine kinase ataxia-telangiectasia mutated and RAD3-related (ATR), with its inhibitor VE-822, induces or increases ICD surrogate markers in the colon cancer model MC38 [91].…”

Autophagy induction by thiostrepton improves the efficacy of immunogenic chemotherapy

“…This implies that some chemotherapeutics can be advantageously combined with immune checkpoint blockers targeting the programmed cell death death 1/ programmed death ligand 1 (PD-1/PD-L1) interaction. [4][5][6][7] Only a fraction of cytotoxicants are able to stimulate ICD, which requires the premortem induction of specific stress pathways, in particular, autophagy and partial endoplasmic reticulum (ER) stress response with the phosphorylation of eukaryotic initiation factor 2α (eIF2α). [8][9][10] Autophagy is required for the lysosomal secretion of ATP, a chemotactic factor that is released from stressed/ dying cancer cells and attracts dendritic cell precursors into the tumor bed via the action on purinergic receptors and may contribute to local inflammasome activation as well.…”

A fluorescent biosensor-based platform for the discovery of immunogenic cancer cell death inducers

“…In this screening campaign, we were able to show that crizotinib acts off-target to induce all ICD hallmarks in human and mouse cancer cell lines if this tyrosine kinase inhibitor was used at a high-dose (10 µM) that trespasses the specific inhibition of its usual target kinases (ALK and ROS1) and hence induces 'off-target' effects on other tyrosine kinases, converging crizotinib de facto into a multikinase inhibitor. High-dose crizotinib induced the redistribution of microtubule-associated protein 1A/1B light chain 3B (hereafter referred to as LC3) fused to green fluorescent protein (GFP) to cytoplasmic dots (LC3-GFP puncta) as a sign of autophagy 39 and stimulated the phosphorylation of eukaryotic translation initiation factor 2α (eIF2α) by the ER stress kinase eIF2α kinase 3 (EIF2AK3). 39 As a correlate of these premortem stress responses, crizotinib stimulated the release of ATP from cancer cells (downstream of autophagy), as well as the surface exposure of CALR (downstream of ER stress).…”

“…High-dose crizotinib induced the redistribution of microtubule-associated protein 1A/1B light chain 3B (hereafter referred to as LC3) fused to green fluorescent protein (GFP) to cytoplasmic dots (LC3-GFP puncta) as a sign of autophagy 39 and stimulated the phosphorylation of eukaryotic translation initiation factor 2α (eIF2α) by the ER stress kinase eIF2α kinase 3 (EIF2AK3). 39 As a correlate of these premortem stress responses, crizotinib stimulated the release of ATP from cancer cells (downstream of autophagy), as well as the surface exposure of CALR (downstream of ER stress). Moreover, crizotinib was highly effective in stimulating the release of HMGB1 and annexin A1 (as a result of cell death) and the activation of a transcriptional program involving the upregulation of mRNAs coding for type-1 interferons and programmed deathligand 1 (PD-L1).…”

“…Moreover, crizotinib was highly effective in stimulating the release of HMGB1 and annexin A1 (as a result of cell death) and the activation of a transcriptional program involving the upregulation of mRNAs coding for type-1 interferons and programmed deathligand 1 (PD-L1). 39 These in vitro effects were particularly strong when crizotinib was combined with non-ICD inducing chemotherapeutics (in particular cisplatin and mitomycin C). Indeed, crizotinib could be advantageously combined with cisplatin in vivo, in preclinical mouse models of non-small cell lung cancer (NSLCL) to induce therapeutic anticancer immune responses.…”