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An Optimized GD2-Targeting Retroviral Cassette for More Potent and Safer Cellular Therapy of Neuroblastoma and Other Cancers.

Thomas S, Straathof K, Himoudi N, Anderson J, Pule M - PLoS ONE (2016)

Bottom Line: GD2 is also expressed in a range of other cancer but with the exception of some peripheral nerves is largely absent from non-transformed tissues.Chimeric Antigen Receptors (CARs) are artificial type I proteins which graft the specificity of a monoclonal antibody onto a T-cell.Here, we describe a GD2-targeting CAR retroviral cassette, which has been optimized for CAR T-cell persistence, efficacy and safety.

View Article: PubMed Central - PubMed

Affiliation: Cancer Institute, University College London, London, United Kingdom.

ABSTRACT
Neuroblastoma is the commonest extra cranial solid cancer of childhood. Despite escalation of treatment regimens, a significant minority of patients die of their disease. Disialoganglioside (GD2) is consistently expressed at high-levels in neuroblastoma tumors, which have been targeted with some success using therapeutic monoclonal antibodies. GD2 is also expressed in a range of other cancer but with the exception of some peripheral nerves is largely absent from non-transformed tissues. Chimeric Antigen Receptors (CARs) are artificial type I proteins which graft the specificity of a monoclonal antibody onto a T-cell. Clinical data with early CAR designs directed against GD2 have shown some promise in Neuroblastoma. Here, we describe a GD2-targeting CAR retroviral cassette, which has been optimized for CAR T-cell persistence, efficacy and safety.

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Related in: MedlinePlus

Consequence and function of iCasp9.The SAR codon-optimized cassette was taken further and compared with the original cassette without iCasp9. (a) Expression of the CAR was unchanged. Depletion is shown by facs after addition of CID. The function of cassettes with and without iCasp9 were assessed by (b) Killing (c) IFN-γ release and (d) IL-2 release. Data shown as means +/-SEM from 4 independent experiments with different donors.
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pone.0152196.g005: Consequence and function of iCasp9.The SAR codon-optimized cassette was taken further and compared with the original cassette without iCasp9. (a) Expression of the CAR was unchanged. Depletion is shown by facs after addition of CID. The function of cassettes with and without iCasp9 were assessed by (b) Killing (c) IFN-γ release and (d) IL-2 release. Data shown as means +/-SEM from 4 independent experiments with different donors.

Mentions: Hence, in terms of expression, stability and response to CID, the codon-optimized SAR-containing receptor (iCasp-HuK) performed the best and this receptor was compared to the original HuK666 CAR for its capacity to mediate cytotoxicity and cytokine release in response to Lan1 cells (Fig 5). Transduced PBMCs were incubated for 72 hours in the absence or presence of 10nm CID and then co-cultured with Lan-1 or A204 cells. An example of the expression of these CARs in the presence and absence of CID is shown in Fig 5A. Levels of HuK666-iCasp9 expression were effectively eliminated by induction of the iCasp9 (Fig 5A).


An Optimized GD2-Targeting Retroviral Cassette for More Potent and Safer Cellular Therapy of Neuroblastoma and Other Cancers.

Thomas S, Straathof K, Himoudi N, Anderson J, Pule M - PLoS ONE (2016)

Consequence and function of iCasp9.The SAR codon-optimized cassette was taken further and compared with the original cassette without iCasp9. (a) Expression of the CAR was unchanged. Depletion is shown by facs after addition of CID. The function of cassettes with and without iCasp9 were assessed by (b) Killing (c) IFN-γ release and (d) IL-2 release. Data shown as means +/-SEM from 4 independent experiments with different donors.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4816271&req=5

pone.0152196.g005: Consequence and function of iCasp9.The SAR codon-optimized cassette was taken further and compared with the original cassette without iCasp9. (a) Expression of the CAR was unchanged. Depletion is shown by facs after addition of CID. The function of cassettes with and without iCasp9 were assessed by (b) Killing (c) IFN-γ release and (d) IL-2 release. Data shown as means +/-SEM from 4 independent experiments with different donors.
Mentions: Hence, in terms of expression, stability and response to CID, the codon-optimized SAR-containing receptor (iCasp-HuK) performed the best and this receptor was compared to the original HuK666 CAR for its capacity to mediate cytotoxicity and cytokine release in response to Lan1 cells (Fig 5). Transduced PBMCs were incubated for 72 hours in the absence or presence of 10nm CID and then co-cultured with Lan-1 or A204 cells. An example of the expression of these CARs in the presence and absence of CID is shown in Fig 5A. Levels of HuK666-iCasp9 expression were effectively eliminated by induction of the iCasp9 (Fig 5A).

Bottom Line: GD2 is also expressed in a range of other cancer but with the exception of some peripheral nerves is largely absent from non-transformed tissues.Chimeric Antigen Receptors (CARs) are artificial type I proteins which graft the specificity of a monoclonal antibody onto a T-cell.Here, we describe a GD2-targeting CAR retroviral cassette, which has been optimized for CAR T-cell persistence, efficacy and safety.

View Article: PubMed Central - PubMed

Affiliation: Cancer Institute, University College London, London, United Kingdom.

ABSTRACT
Neuroblastoma is the commonest extra cranial solid cancer of childhood. Despite escalation of treatment regimens, a significant minority of patients die of their disease. Disialoganglioside (GD2) is consistently expressed at high-levels in neuroblastoma tumors, which have been targeted with some success using therapeutic monoclonal antibodies. GD2 is also expressed in a range of other cancer but with the exception of some peripheral nerves is largely absent from non-transformed tissues. Chimeric Antigen Receptors (CARs) are artificial type I proteins which graft the specificity of a monoclonal antibody onto a T-cell. Clinical data with early CAR designs directed against GD2 have shown some promise in Neuroblastoma. Here, we describe a GD2-targeting CAR retroviral cassette, which has been optimized for CAR T-cell persistence, efficacy and safety.

Show MeSH
Related in: MedlinePlus