Choice of costimulatory domains and of cytokines determines CAR T-cell activity in neuroblastoma

Research output: Contribution to journalArticle

Abstract

Chimeric antigen receptor (CAR) T-cell therapy has been shown to be dramatically effective in the treatment of B-cell malignancies. However, there are still substantial obstacles to overcome, before similar responses can be achieved in patients with solid tumors. We evaluated both in vitro and in a preclinical murine model the efficacy of different 2nd and 3rd generation CAR constructs targeting GD2, a disial-ganglioside expressed on the surface of neuroblastoma (NB) tumor cells. In order to address potential safety concerns regarding clinical application, an inducible safety switch, namely inducible Caspase-9 (iC9), was also included in the vector constructs. Our data indicate that a 3rd generation CAR incorporating CD28.4-1BB costimulatory domains is associated with improved anti-tumor efficacy as compared with a CAR incorporating the combination of CD28.OX40 domains. We demonstrate that the choice of 4-1BB signaling results into significant amelioration of several CAR T-cell characteristics, including: 1) T-cell exhaustion, 2) basal T-cell activation, 3) in vivo tumor control and 4) T-cell persistence. The fine-tuning of T-cell culture conditions obtained using IL7 and IL15 was found to be synergic with the CAR.GD2 design in increasing the anti-tumor activity of CAR T cells. We also demonstrate that activation of the suicide gene iC9, included in our construct without significantly impairing neither CAR expression nor anti-tumor activity, leads to a prompt induction of apoptosis of GD2.CAR T cells. Altogether, these findings are instrumental in optimizing the function of CAR T-cell products to be employed in the treatment of children with NB.

Original languageEnglish
Article numbere1433518
JournalOncoImmunology
Volume7
Issue number6
DOIs
Publication statusPublished - Jun 3 2018

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Antigen Receptors
T-Cell Antigen Receptor
Neuroblastoma
Cytokines
Neoplasms
T-Lymphocytes
Caspase 9
Safety
Interleukin-15
Interleukin-7
Gangliosides
Cell- and Tissue-Based Therapy
Suicide
Transcriptional Activation
B-Lymphocytes
Cell Culture Techniques
Apoptosis
Therapeutics

Keywords

  • CAR.GD2 design
  • CD28.4-1BB costimulatory domains
  • Chimeric antigen receptor (CAR)
  • Neuroblastoma
  • solid tumors
  • T-cell exhaustion

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology
  • Oncology

Cite this

@article{6497176a61584a70a44037efcf3952d6,
title = "Choice of costimulatory domains and of cytokines determines CAR T-cell activity in neuroblastoma",
abstract = "Chimeric antigen receptor (CAR) T-cell therapy has been shown to be dramatically effective in the treatment of B-cell malignancies. However, there are still substantial obstacles to overcome, before similar responses can be achieved in patients with solid tumors. We evaluated both in vitro and in a preclinical murine model the efficacy of different 2nd and 3rd generation CAR constructs targeting GD2, a disial-ganglioside expressed on the surface of neuroblastoma (NB) tumor cells. In order to address potential safety concerns regarding clinical application, an inducible safety switch, namely inducible Caspase-9 (iC9), was also included in the vector constructs. Our data indicate that a 3rd generation CAR incorporating CD28.4-1BB costimulatory domains is associated with improved anti-tumor efficacy as compared with a CAR incorporating the combination of CD28.OX40 domains. We demonstrate that the choice of 4-1BB signaling results into significant amelioration of several CAR T-cell characteristics, including: 1) T-cell exhaustion, 2) basal T-cell activation, 3) in vivo tumor control and 4) T-cell persistence. The fine-tuning of T-cell culture conditions obtained using IL7 and IL15 was found to be synergic with the CAR.GD2 design in increasing the anti-tumor activity of CAR T cells. We also demonstrate that activation of the suicide gene iC9, included in our construct without significantly impairing neither CAR expression nor anti-tumor activity, leads to a prompt induction of apoptosis of GD2.CAR T cells. Altogether, these findings are instrumental in optimizing the function of CAR T-cell products to be employed in the treatment of children with NB.",
keywords = "CAR.GD2 design, CD28.4-1BB costimulatory domains, Chimeric antigen receptor (CAR), Neuroblastoma, solid tumors, T-cell exhaustion",
author = "Concetta Quintarelli and Domenico Orlando and Iolanda Boffa and Marika Guercio and Polito, {Vinicia Assunta} and Andrea Petretto and Chiara Lavarello and Matilde Sinibaldi and Gerrit Weber and {Del Bufalo}, Francesca and Ezio Giorda and Marco Scarsella and Stefania Petrini and Daria Pagliara and Franco Locatelli and {De Angelis}, Biagio and Ignazio Caruana",
year = "2018",
month = "6",
day = "3",
doi = "10.1080/2162402X.2018.1433518",
language = "English",
volume = "7",
journal = "OncoImmunology",
issn = "2162-4011",
publisher = "Taylor and Francis Inc.",
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T1 - Choice of costimulatory domains and of cytokines determines CAR T-cell activity in neuroblastoma

AU - Quintarelli, Concetta

AU - Orlando, Domenico

AU - Boffa, Iolanda

AU - Guercio, Marika

AU - Polito, Vinicia Assunta

AU - Petretto, Andrea

AU - Lavarello, Chiara

AU - Sinibaldi, Matilde

AU - Weber, Gerrit

AU - Del Bufalo, Francesca

AU - Giorda, Ezio

AU - Scarsella, Marco

AU - Petrini, Stefania

AU - Pagliara, Daria

AU - Locatelli, Franco

AU - De Angelis, Biagio

AU - Caruana, Ignazio

PY - 2018/6/3

Y1 - 2018/6/3

N2 - Chimeric antigen receptor (CAR) T-cell therapy has been shown to be dramatically effective in the treatment of B-cell malignancies. However, there are still substantial obstacles to overcome, before similar responses can be achieved in patients with solid tumors. We evaluated both in vitro and in a preclinical murine model the efficacy of different 2nd and 3rd generation CAR constructs targeting GD2, a disial-ganglioside expressed on the surface of neuroblastoma (NB) tumor cells. In order to address potential safety concerns regarding clinical application, an inducible safety switch, namely inducible Caspase-9 (iC9), was also included in the vector constructs. Our data indicate that a 3rd generation CAR incorporating CD28.4-1BB costimulatory domains is associated with improved anti-tumor efficacy as compared with a CAR incorporating the combination of CD28.OX40 domains. We demonstrate that the choice of 4-1BB signaling results into significant amelioration of several CAR T-cell characteristics, including: 1) T-cell exhaustion, 2) basal T-cell activation, 3) in vivo tumor control and 4) T-cell persistence. The fine-tuning of T-cell culture conditions obtained using IL7 and IL15 was found to be synergic with the CAR.GD2 design in increasing the anti-tumor activity of CAR T cells. We also demonstrate that activation of the suicide gene iC9, included in our construct without significantly impairing neither CAR expression nor anti-tumor activity, leads to a prompt induction of apoptosis of GD2.CAR T cells. Altogether, these findings are instrumental in optimizing the function of CAR T-cell products to be employed in the treatment of children with NB.

AB - Chimeric antigen receptor (CAR) T-cell therapy has been shown to be dramatically effective in the treatment of B-cell malignancies. However, there are still substantial obstacles to overcome, before similar responses can be achieved in patients with solid tumors. We evaluated both in vitro and in a preclinical murine model the efficacy of different 2nd and 3rd generation CAR constructs targeting GD2, a disial-ganglioside expressed on the surface of neuroblastoma (NB) tumor cells. In order to address potential safety concerns regarding clinical application, an inducible safety switch, namely inducible Caspase-9 (iC9), was also included in the vector constructs. Our data indicate that a 3rd generation CAR incorporating CD28.4-1BB costimulatory domains is associated with improved anti-tumor efficacy as compared with a CAR incorporating the combination of CD28.OX40 domains. We demonstrate that the choice of 4-1BB signaling results into significant amelioration of several CAR T-cell characteristics, including: 1) T-cell exhaustion, 2) basal T-cell activation, 3) in vivo tumor control and 4) T-cell persistence. The fine-tuning of T-cell culture conditions obtained using IL7 and IL15 was found to be synergic with the CAR.GD2 design in increasing the anti-tumor activity of CAR T cells. We also demonstrate that activation of the suicide gene iC9, included in our construct without significantly impairing neither CAR expression nor anti-tumor activity, leads to a prompt induction of apoptosis of GD2.CAR T cells. Altogether, these findings are instrumental in optimizing the function of CAR T-cell products to be employed in the treatment of children with NB.

KW - CAR.GD2 design

KW - CD28.4-1BB costimulatory domains

KW - Chimeric antigen receptor (CAR)

KW - Neuroblastoma

KW - solid tumors

KW - T-cell exhaustion

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