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 language | English |
|---|---|
| Article number | e1433518 |
| Journal | OncoImmunology |
| Volume | 7 |
| Issue number | 6 |
| DOIs | |
| Publication status | Published - Jun 3 2018 |
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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
Choice of costimulatory domains and of cytokines determines CAR T-cell activity in neuroblastoma. / Quintarelli, Concetta; Orlando, Domenico; Boffa, Iolanda; Guercio, Marika; Polito, Vinicia Assunta; Petretto, Andrea; Lavarello, Chiara; Sinibaldi, Matilde; Weber, Gerrit; Del Bufalo, Francesca; Giorda, Ezio; Scarsella, Marco; Petrini, Stefania; Pagliara, Daria; Locatelli, Franco; De Angelis, Biagio; Caruana, Ignazio.
In: OncoImmunology, Vol. 7, No. 6, e1433518, 03.06.2018.Research output: Contribution to journal › Article
}
TY - JOUR
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
UR - http://www.scopus.com/inward/record.url?scp=85044044943&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85044044943&partnerID=8YFLogxK
U2 - 10.1080/2162402X.2018.1433518
DO - 10.1080/2162402X.2018.1433518
M3 - Article
AN - SCOPUS:85044044943
VL - 7
JO - OncoImmunology
JF - OncoImmunology
SN - 2162-4011
IS - 6
M1 - e1433518
ER -