Anticancer chemotherapy and radiotherapy trigger both non-cell-autonomous and cell-autonomous death

Isabelle Martins,Frédéric Law,Syed Qasim Raza,Delphine Dugué,David M Ojcius,Guido Kroemer,Dora Sabino,Nazanine Modjtahedi,Dorine De Jong,Laurent Voisin,Fanny Jaulin,Awatef Allouch,Elodie Mintet,Éric Deutsch ,Marie‐Lise Gougeon ,Haithem Dakhli,Catherine Brenner,Maxime Thoreau,Mauro Piacentini,Pascale Bertrand,Jean-Luc Perfettini

doi:10.1038/s41419-018-0747-y

Abstract

Even though cell death modalities elicited by anticancer chemotherapy and radiotherapy have been extensively studied, the ability of anticancer treatments to induce non-cell-autonomous death has never been investigated. By means of multispectral imaging flow-cytometry-based technology, we analyzed the lethal fate of cancer cells that were treated with conventional anticancer agents and co-cultured with untreated cells, observing that anticancer agents can simultaneously trigger cell-autonomous and non-cell-autonomous death in treated and untreated cells. After ionizing radiation, oxaliplatin, or cisplatin treatment, fractions of treated cancer cell populations were eliminated through cell-autonomous death mechanisms, while other fractions of the treated cancer cells engulfed and killed neighboring cells through non-cell-autonomous processes, including cellular cannibalism. Under conditions of treatment with paclitaxel, non-cell-autonomous and cell-autonomous death were both detected in the treated cell population, while untreated neighboring cells exhibited features of apoptotic demise. The transcriptional activity of p53 tumor-suppressor protein contributed to the execution of cell-autonomous death, yet failed to affect the non-cell-autonomous death by cannibalism for the majority of tested anticancer agents, indicating that the induction of non-cell-autonomous death can occur under conditions in which cell-autonomous death was impaired. Altogether, these results reveal that chemotherapy and radiotherapy can induce both non-cell-autonomous and cell-autonomous death of cancer cells, highlighting the heterogeneity of cell death responses to anticancer treatments and the unsuspected potential contribution of non-cell-autonomous death to the global effects of anticancer treatment.

Highlights

From the initial discovery of programmed cell death during animal development[1] to the recent identification of entotic death during embryo implantation[2], a cornucopiaOfficial journal of the Cell Death Differentiation AssociationMartins et al Cell Death and Disease (2018)9:716 catabolic activities (involving caspases, receptorinteracting protein kinases (RIPKs), mixed lineage kinase domain-like proteins, or cathepsins), and in relation to their ability to elicit an inflammatory reaction or to stimulate an immune response
To precisely analyze the cellular mechanisms through which cancer cells may simultaneously undergo direct and bystander cell killing in response to Ionizing radiation (IR), we designed a novel cell death profiling assay based on co-culture of untreated cancer cells that have been labeled with the red fluorescent probe 5-(and-6)-(((4-chloromethyl)benzoyl) amino) tetramethylrhodamine (CMTMR) and irradiated isogenic cancer cells that have been labeled with the green fluorescent probe 5-chloromethylfluorescein diacetate (CMFDA)
The simultaneous detection of the above-indicated parameters allowed us to detect, through the use of multispectral imaging flow cytometry, the execution of at least four types of cell death on target cells and on neighboring cells (Fig. 1b), discriminating cell-autonomous death (CAD) from non-cell-autonomous death (NCAD) and direct cell killing from bystander lethal effects

Summary

Introduction

From the initial discovery of programmed cell death during animal development[1] to the recent identification of entotic death during embryo implantation[2], a cornucopiaOfficial journal of the Cell Death Differentiation AssociationMartins et al Cell Death and Disease (2018)9:716 catabolic activities (involving (or not) caspases, receptorinteracting protein kinases (RIPKs), mixed lineage kinase domain-like proteins, or cathepsins), and in relation to their ability to elicit an inflammatory reaction or to stimulate an immune response. A classification of cell death modalities built on these criteria has been proposed[5] and led to the ordering of lethal processes into three distinct types: type I cell death (or apoptosis), type II cell death (or autophagic cell death), and type III cell death (or necrosis) All these processes, which are executed in a cellautonomous manner, can be induced in the targeted stressed cells or at a distance, in the neighboring cells (through bystander effects). Additional cell death mechanisms (such as entosis or emperitosis) have been described and associated with this neglected subgroup of cell death modalities[7, 8] Their examination revealed the existence of cell death processes that are elicited after the engulfment of live cells by neighboring live cells. We recently defined NCAD as type IV cell death[6]

Methods

Results

Conclusion