Cytokine-Induced Killer Cells Kill Chemo-surviving Melanoma Cancer Stem Cells.
Purpose:The MHC-unrestricted activity of cytokine-induced killer (CIK) cells against chemo-surviving melanoma cancer stem cells (mCSC) was explored, as CSCs are considered responsible for chemoresistance and relapses.Experimental Design:Putative mCSCs were visualized by engineering patient-derived melanoma cells (MC) with a lentiviral vector encoding eGFP under expression control by stemness gene promoteroct4Their stemness potential was confirmedin vivoby limiting dilution assays. We explored the sensitivity of eGFP+mCSCs to chemotherapy (CHT), BRAF inhibitor (BRAFi) or CIK cells, as single agents or in sequence,in vitroFirst, we treated MCsin vitrowith fotemustine or dabrafenib (BRAF-mutated cases); then, surviving MCs, enriched in mCSCs, were challenged with autologous CIK cells. CIK cell activity against chemoresistant mCSCs was confirmedin vivoin two distinct immunodeficient murine models.Results:We visualized eGFP+mCSCs (14% ± 2.1%) in 11 MCs. The tumorigenic precursor ratein vivowas higher within eGFP+MCs (1/42) compared with the eGFP-counterpart (1/4,870).In vitromCSCs were relatively resistant to CHT and BRAFi, but killed by CIK cells (n= 11, 8/11 autologous), with specific lysis ranging from 95% [effector:tumor ratio (E:T), 40:1] to 20% (E:T 1:3).In vivoinfusion of autologous CIK cells into mice bearing xenografts from three distinct melanomas demonstrated significant tumor responses involving CHT-spared eGFP+mCSCs (P= 0.001). Sequential CHT-immunotherapy treatment retained antitumor activity (n= 12,P= 0.001) reducing mCSC rates (P= 0.01).Conclusions:These findings are the first demonstration that immunotherapy with CIK cells is active against autologous mCSCs surviving CHT or BRAFi. An experimental platform for mCSC study and rationale for CIK cells in melanoma clinical study is provided.Clin Cancer Res; 23(9); 2277-88. ©2016 AACR.
Division of Medical Oncology, Experimental Cell Therapy, Candiolo Cancer Institute, FPO-IRCCS, Candiolo, Torino, Italy.
Loretta
L
Gammaitoni
Division of Medical Oncology, Experimental Cell Therapy, Candiolo Cancer Institute, FPO-IRCCS, Candiolo, Torino, Italy.
Lidia
L
Giraudo
Division of Medical Oncology, Experimental Cell Therapy, Candiolo Cancer Institute, FPO-IRCCS, Candiolo, Torino, Italy.|Department of Oncology, University of Torino, Candiolo, Torino, Italy.
Marco
M
Macagno
Division of Medical Oncology, Experimental Cell Therapy, Candiolo Cancer Institute, FPO-IRCCS, Candiolo, Torino, Italy.|Department of Oncology, University of Torino, Candiolo, Torino, Italy.
Valeria
V
Leuci
Division of Medical Oncology, Experimental Cell Therapy, Candiolo Cancer Institute, FPO-IRCCS, Candiolo, Torino, Italy.
Giulia
G
Mesiano
Division of Medical Oncology, Experimental Cell Therapy, Candiolo Cancer Institute, FPO-IRCCS, Candiolo, Torino, Italy.|Department of Oncology, University of Torino, Candiolo, Torino, Italy.
Ramona
R
Rotolo
Laboratory of Translational Cancer Medicine, Candiolo Cancer Institute, FPO-IRCCS, Candiolo, Torino, Italy.
Francesco
F
Sassi
Division of Medical Oncology, Experimental Cell Therapy, Candiolo Cancer Institute, FPO-IRCCS, Candiolo, Torino, Italy.|Department of Oncology, University of Torino, Candiolo, Torino, Italy.|Section of Dermatology, Department of Medical Sciences, University of Torino, Torino, Italy.
Martina
M
Sanlorenzo
Surgical Dermatology, Candiolo Cancer Institute, FPO-IRCCS, Candiolo, Torino, Italy.
Alessandro
A
Zaccagna
Pathology, Candiolo Cancer Institute, FPO-IRCCS, Candiolo, Torino, Italy.
Alberto
A
Pisacane
Pathology, Candiolo Cancer Institute, FPO-IRCCS, Candiolo, Torino, Italy.
Rebecca
R
Senetta
Department of Oncology, University of Torino, Candiolo, Torino, Italy.
Michela
M
Cangemi
Division of Medical Oncology, Experimental Cell Therapy, Candiolo Cancer Institute, FPO-IRCCS, Candiolo, Torino, Italy.|Department of Oncology, University of Torino, Candiolo, Torino, Italy.
Giulia
G
Cattaneo
Division of Medical Oncology, Experimental Cell Therapy, Candiolo Cancer Institute, FPO-IRCCS, Candiolo, Torino, Italy.
Valentina
V
Martin
Division of Medical Oncology, Experimental Cell Therapy, Candiolo Cancer Institute, FPO-IRCCS, Candiolo, Torino, Italy.
Valentina
V
Coha
Division of Medical Oncology, Experimental Cell Therapy, Candiolo Cancer Institute, FPO-IRCCS, Candiolo, Torino, Italy.
Susanna
S
Gallo
Division of Medical Oncology, Experimental Cell Therapy, Candiolo Cancer Institute, FPO-IRCCS, Candiolo, Torino, Italy.|Laboratory of Translational Cancer Medicine, Candiolo Cancer Institute, FPO-IRCCS, Candiolo, Torino, Italy.
Ymera
Y
Pignochino
Pathology, Candiolo Cancer Institute, FPO-IRCCS, Candiolo, Torino, Italy.
Anna
A
Sapino
Division of Medical Oncology, Experimental Cell Therapy, Candiolo Cancer Institute, FPO-IRCCS, Candiolo, Torino, Italy.
Giovanni
G
Grignani
Division of Medical Oncology, Experimental Cell Therapy, Candiolo Cancer Institute, FPO-IRCCS, Candiolo, Torino, Italy.
Fabrizio
F
Carnevale-Schianca
Division of Medical Oncology, Experimental Cell Therapy, Candiolo Cancer Institute, FPO-IRCCS, Candiolo, Torino, Italy.|Department of Oncology, University of Torino, Candiolo, Torino, Italy.
Massimo
M
Aglietta
Division of Medical Oncology, Experimental Cell Therapy, Candiolo Cancer Institute, FPO-IRCCS, Candiolo, Torino, Italy. dario.sangiolo@unito.it.|Department of Oncology, University of Torino, Candiolo, Torino, Italy.
Dario
D
Sangiolo
D000818:Animals; D063646:Carcinogenesis / Q000235:genetics / Q000276:immunology; D045744:Cell Line, Tumor; D055612:Cytokine-Induced Killer Cells / Q000276:immunology / Q000637:transplantation*; D003602:Cytotoxicity, Immunologic; D019008:Drug Resistance, Neoplasm / Q000235:genetics / Q000276:immunology; D006801:Humans; D007093:Imidazoles / Q000008:administration & dosage; D016219:Immunotherapy, Adoptive / Q000379:methods*; D016086:Lentivirus / Q000235:genetics; D008545:Melanoma / Q000188:drug therapy / Q000276:immunology / Q000473:pathology / Q000628:therapy*; D051379:Mice; D009364:Neoplasm Recurrence, Local; D014411:Neoplastic Stem Cells / Q000276:immunology / Q000637:transplantation; D009607:Nitrosourea Compounds / Q000008:administration & dosage; D009943:Organophosphorus Compounds / Q000008:administration & dosage; D010091:Oximes / Q000008:administration & dosage; D048493:Proto-Oncogene Proteins B-raf / Q000037:antagonists & inhibitors / Q000235:genetics; D023041:Xenograft Model Antitumor Assays
D007093:Imidazoles; D009607:Nitrosourea Compounds; D009943:Organophosphorus Compounds; D010091:Oximes; C482119:BRAF protein, human; D048493:Proto-Oncogene Proteins B-raf; C054368:fotemustine; C561627:dabrafenib