Cargando…

GD2 redirected CAR T and activated NK-cell-mediated secretion of IFNγ overcomes MYCN-dependent IDO1 inhibition, contributing to neuroblastoma cell immune escape

Immune escape mechanisms employed by neuroblastoma (NB) cells include secretion of immunosuppressive factors disrupting effective antitumor immunity. The use of cellular therapy to treat solid tumors needs to be implemented. Killing activity of anti-GD2 Chimeric Antigen Receptor (CAR) T or natural k...

Descripción completa

Detalles Bibliográficos
Autores principales: Caforio, Matteo, Sorino, Cristina, Caruana, Ignazio, Weber, Gerrit, Camera, Antonio, Cifaldi, Loredana, De Angelis, Biagio, Del Bufalo, Francesca, Vitale, Alessia, Goffredo, Bianca Maria, De Vito, Rita, Fruci, Doriana, Quintarelli, Concetta, Fanciulli, Maurizio, Locatelli, Franco, Folgiero, Valentina
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BMJ Publishing Group 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7978286/
https://www.ncbi.nlm.nih.gov/pubmed/33737337
http://dx.doi.org/10.1136/jitc-2020-001502
_version_ 1783667179700355072
author Caforio, Matteo
Sorino, Cristina
Caruana, Ignazio
Weber, Gerrit
Camera, Antonio
Cifaldi, Loredana
De Angelis, Biagio
Del Bufalo, Francesca
Vitale, Alessia
Goffredo, Bianca Maria
De Vito, Rita
Fruci, Doriana
Quintarelli, Concetta
Fanciulli, Maurizio
Locatelli, Franco
Folgiero, Valentina
author_facet Caforio, Matteo
Sorino, Cristina
Caruana, Ignazio
Weber, Gerrit
Camera, Antonio
Cifaldi, Loredana
De Angelis, Biagio
Del Bufalo, Francesca
Vitale, Alessia
Goffredo, Bianca Maria
De Vito, Rita
Fruci, Doriana
Quintarelli, Concetta
Fanciulli, Maurizio
Locatelli, Franco
Folgiero, Valentina
author_sort Caforio, Matteo
collection PubMed
description Immune escape mechanisms employed by neuroblastoma (NB) cells include secretion of immunosuppressive factors disrupting effective antitumor immunity. The use of cellular therapy to treat solid tumors needs to be implemented. Killing activity of anti-GD2 Chimeric Antigen Receptor (CAR) T or natural killer (NK) cells against target NB cells was assessed through coculture experiments and quantified by FACS analysis. ELISA assay was used to quantify interferon-γ (IFNγ) secreted by NK and CAR T cells. Real Time PCR and Western Blot were performed to analyze gene and protein levels modifications. Transcriptional study was performed by chromatin immunoprecipitation and luciferase reporter assays on experiments of mutagenesis on the promoter sequence. NB tissue sample were analyzed by IHC and Real Time PCR to perform correlation study. We demonstrate that Indoleamine-pyrrole 2,3-dioxygenase1 (IDO1), due to its ability to convert tryptophan into kynurenines, is involved in NB resistance to activity of immune cells. In NB, IDO1 is able to inhibit the anti-tumor effect displayed by of both anti-GD2 CAR (GD2.CAR) T-cell and NK cells, mainly by impairing their IFNγ production. Furthermore, inhibition of MYCN expression in NB results into accumulation of IDO1 and consequently of kynurenines, which negatively affect the immune surveillance. Inverse correlation between IDO1 and MYCN expression has been observed in a wide cohort of NB samples. This finding was supported by the identification of a transcriptional repressive role of MYCN on IDO1 promoter. The evidence of IDO1 involvement in NB immune escape and its ability to impair NK and GD2.CAR T-cell activity contribute to clarify one of the possible mechanisms responsible for the limited efficacy of these immunotherapeutic approaches. A combined therapy of NK or GD2.CAR T-cells with IDO1 inhibitors, a class of compounds already in phase I/II clinical studies, could represent a new and still unexplored strategy capable to improve long-term efficacy of these immunotherapeutic approaches.
format Online
Article
Text
id pubmed-7978286
institution National Center for Biotechnology Information
language English
publishDate 2021
publisher BMJ Publishing Group
record_format MEDLINE/PubMed
spelling pubmed-79782862021-03-30 GD2 redirected CAR T and activated NK-cell-mediated secretion of IFNγ overcomes MYCN-dependent IDO1 inhibition, contributing to neuroblastoma cell immune escape Caforio, Matteo Sorino, Cristina Caruana, Ignazio Weber, Gerrit Camera, Antonio Cifaldi, Loredana De Angelis, Biagio Del Bufalo, Francesca Vitale, Alessia Goffredo, Bianca Maria De Vito, Rita Fruci, Doriana Quintarelli, Concetta Fanciulli, Maurizio Locatelli, Franco Folgiero, Valentina J Immunother Cancer Immunotherapy Biomarkers Immune escape mechanisms employed by neuroblastoma (NB) cells include secretion of immunosuppressive factors disrupting effective antitumor immunity. The use of cellular therapy to treat solid tumors needs to be implemented. Killing activity of anti-GD2 Chimeric Antigen Receptor (CAR) T or natural killer (NK) cells against target NB cells was assessed through coculture experiments and quantified by FACS analysis. ELISA assay was used to quantify interferon-γ (IFNγ) secreted by NK and CAR T cells. Real Time PCR and Western Blot were performed to analyze gene and protein levels modifications. Transcriptional study was performed by chromatin immunoprecipitation and luciferase reporter assays on experiments of mutagenesis on the promoter sequence. NB tissue sample were analyzed by IHC and Real Time PCR to perform correlation study. We demonstrate that Indoleamine-pyrrole 2,3-dioxygenase1 (IDO1), due to its ability to convert tryptophan into kynurenines, is involved in NB resistance to activity of immune cells. In NB, IDO1 is able to inhibit the anti-tumor effect displayed by of both anti-GD2 CAR (GD2.CAR) T-cell and NK cells, mainly by impairing their IFNγ production. Furthermore, inhibition of MYCN expression in NB results into accumulation of IDO1 and consequently of kynurenines, which negatively affect the immune surveillance. Inverse correlation between IDO1 and MYCN expression has been observed in a wide cohort of NB samples. This finding was supported by the identification of a transcriptional repressive role of MYCN on IDO1 promoter. The evidence of IDO1 involvement in NB immune escape and its ability to impair NK and GD2.CAR T-cell activity contribute to clarify one of the possible mechanisms responsible for the limited efficacy of these immunotherapeutic approaches. A combined therapy of NK or GD2.CAR T-cells with IDO1 inhibitors, a class of compounds already in phase I/II clinical studies, could represent a new and still unexplored strategy capable to improve long-term efficacy of these immunotherapeutic approaches. BMJ Publishing Group 2021-03-18 /pmc/articles/PMC7978286/ /pubmed/33737337 http://dx.doi.org/10.1136/jitc-2020-001502 Text en © Author(s) (or their employer(s)) 2021. Re-use permitted under CC BY-NC. No commercial re-use. See rights and permissions. Published by BMJ. http://creativecommons.org/licenses/by-nc/4.0/ http://creativecommons.org/licenses/by-nc/4.0/This is an open access article distributed in accordance with the Creative Commons Attribution Non Commercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited, appropriate credit is given, any changes made indicated, and the use is non-commercial. See http://creativecommons.org/licenses/by-nc/4.0/.
spellingShingle Immunotherapy Biomarkers
Caforio, Matteo
Sorino, Cristina
Caruana, Ignazio
Weber, Gerrit
Camera, Antonio
Cifaldi, Loredana
De Angelis, Biagio
Del Bufalo, Francesca
Vitale, Alessia
Goffredo, Bianca Maria
De Vito, Rita
Fruci, Doriana
Quintarelli, Concetta
Fanciulli, Maurizio
Locatelli, Franco
Folgiero, Valentina
GD2 redirected CAR T and activated NK-cell-mediated secretion of IFNγ overcomes MYCN-dependent IDO1 inhibition, contributing to neuroblastoma cell immune escape
title GD2 redirected CAR T and activated NK-cell-mediated secretion of IFNγ overcomes MYCN-dependent IDO1 inhibition, contributing to neuroblastoma cell immune escape
title_full GD2 redirected CAR T and activated NK-cell-mediated secretion of IFNγ overcomes MYCN-dependent IDO1 inhibition, contributing to neuroblastoma cell immune escape
title_fullStr GD2 redirected CAR T and activated NK-cell-mediated secretion of IFNγ overcomes MYCN-dependent IDO1 inhibition, contributing to neuroblastoma cell immune escape
title_full_unstemmed GD2 redirected CAR T and activated NK-cell-mediated secretion of IFNγ overcomes MYCN-dependent IDO1 inhibition, contributing to neuroblastoma cell immune escape
title_short GD2 redirected CAR T and activated NK-cell-mediated secretion of IFNγ overcomes MYCN-dependent IDO1 inhibition, contributing to neuroblastoma cell immune escape
title_sort gd2 redirected car t and activated nk-cell-mediated secretion of ifnγ overcomes mycn-dependent ido1 inhibition, contributing to neuroblastoma cell immune escape
topic Immunotherapy Biomarkers
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7978286/
https://www.ncbi.nlm.nih.gov/pubmed/33737337
http://dx.doi.org/10.1136/jitc-2020-001502
work_keys_str_mv AT caforiomatteo gd2redirectedcartandactivatednkcellmediatedsecretionofifngovercomesmycndependentido1inhibitioncontributingtoneuroblastomacellimmuneescape
AT sorinocristina gd2redirectedcartandactivatednkcellmediatedsecretionofifngovercomesmycndependentido1inhibitioncontributingtoneuroblastomacellimmuneescape
AT caruanaignazio gd2redirectedcartandactivatednkcellmediatedsecretionofifngovercomesmycndependentido1inhibitioncontributingtoneuroblastomacellimmuneescape
AT webergerrit gd2redirectedcartandactivatednkcellmediatedsecretionofifngovercomesmycndependentido1inhibitioncontributingtoneuroblastomacellimmuneescape
AT cameraantonio gd2redirectedcartandactivatednkcellmediatedsecretionofifngovercomesmycndependentido1inhibitioncontributingtoneuroblastomacellimmuneescape
AT cifaldiloredana gd2redirectedcartandactivatednkcellmediatedsecretionofifngovercomesmycndependentido1inhibitioncontributingtoneuroblastomacellimmuneescape
AT deangelisbiagio gd2redirectedcartandactivatednkcellmediatedsecretionofifngovercomesmycndependentido1inhibitioncontributingtoneuroblastomacellimmuneescape
AT delbufalofrancesca gd2redirectedcartandactivatednkcellmediatedsecretionofifngovercomesmycndependentido1inhibitioncontributingtoneuroblastomacellimmuneescape
AT vitalealessia gd2redirectedcartandactivatednkcellmediatedsecretionofifngovercomesmycndependentido1inhibitioncontributingtoneuroblastomacellimmuneescape
AT goffredobiancamaria gd2redirectedcartandactivatednkcellmediatedsecretionofifngovercomesmycndependentido1inhibitioncontributingtoneuroblastomacellimmuneescape
AT devitorita gd2redirectedcartandactivatednkcellmediatedsecretionofifngovercomesmycndependentido1inhibitioncontributingtoneuroblastomacellimmuneescape
AT frucidoriana gd2redirectedcartandactivatednkcellmediatedsecretionofifngovercomesmycndependentido1inhibitioncontributingtoneuroblastomacellimmuneescape
AT quintarelliconcetta gd2redirectedcartandactivatednkcellmediatedsecretionofifngovercomesmycndependentido1inhibitioncontributingtoneuroblastomacellimmuneescape
AT fanciullimaurizio gd2redirectedcartandactivatednkcellmediatedsecretionofifngovercomesmycndependentido1inhibitioncontributingtoneuroblastomacellimmuneescape
AT locatellifranco gd2redirectedcartandactivatednkcellmediatedsecretionofifngovercomesmycndependentido1inhibitioncontributingtoneuroblastomacellimmuneescape
AT folgierovalentina gd2redirectedcartandactivatednkcellmediatedsecretionofifngovercomesmycndependentido1inhibitioncontributingtoneuroblastomacellimmuneescape