T cell receptor gene therapy: strategies for optimizing transgenic TCR pairing

Govers, Coen; Sebestyén, Zsolt; Coccoris, Miriam; Willemsen, Ralph A.; Debets, Reno

doi:10.1016/j.molmed.2009.12.004

Cited by 92 publications

(80 citation statements)

References 84 publications

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“…This method of TCR introduction results in T-cells which can be forced to target specific antigens expressed only by cancer cells. [5][6][7][8] To mimic the TCR function, antibodies and single-chain variable fragments (scFv) were produced showing specific binding and cell kill. Previously, Willemsen et al 5 demonstrated T-cell targeting to melanoma cells using the G8 Fab fragment.…”

Section: Introductionmentioning

confidence: 99%

Targeting melanoma with immunoliposomes coupled to anti-MAGE A1 TCR-like single-chain antibody

Saeed

Brakel

Zalba

et al. 2016

IJN

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Therapy of melanoma using T-cells with genetically introduced T-cell receptors (TCRs) directed against a tumor-selective cancer testis antigen (CTA) NY-ESO1 demonstrated clear antitumor responses in patients without side effects. Here, we exploited the concept of TCR-mediated targeting through introduction of single-chain variable fragment (scFv) antibodies that mimic TCRs in binding major histocompatibility complex-restricted CTA. We produced scFv antibodies directed against Melanoma AntiGEn A1 (MAGE A1) presented by human leukocyte antigen A1 (HLA-A1), in short M1/A1, and coupled these TCR-like antibodies to liposomes to achieve specific melanoma targeting. Two anti-M1/A1 antibodies with different ligand-binding affinities were derived from a phage-display library and reformatted into scFvs with an added cysteine at their carboxyl termini. Protein production conditions, ie, bacterial strain, temperature, time, and compartments, were optimized, and following production, scFv proteins were purified by immobilized metal ion affinity chromatography. Batches of pure scFvs were validated for specific binding to M1/A1-positive B-cells by flow cytometry. Coupling of scFvs to liposomes was conducted by employing different conditions, and an optimized procedure was achieved. In vitro experiments with immunoliposomes demonstrated binding of M1/A1-positive B-cells as well as M1/A1-positive melanoma cells and internalization by these cells using flow cytometry and confocal microscopy. Notably, the scFv with nonenhanced affinity of M1/A1, but not the one with enhanced affinity, was exclusively bound to and internalized by melanoma tumor cells expressing M1/A1. Taken together, antigen-mediated targeting of tumor cells as well as promoting internalization of nanoparticles by these tumor cells is mediated by TCR-like scFv and can contribute to melanoma-specific targeting.

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Section: Introductionmentioning

confidence: 99%

Targeting melanoma with immunoliposomes coupled to anti-MAGE A1 TCR-like single-chain antibody

Saeed

Brakel

Zalba

et al. 2016

IJN

Self Cite

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“…Current clinical data suggest that the increased functional avidity of the transduced cells may be associated with an improved clinical objective response (41). Previous studies have dealt with improving the functional avidity of TCR-transduced cells using structural modifications of mainly the extracellular parts of the TCR constant regions (10,42). In this study, we focused essentially on the TCR TM region and aimed at enhancing TCR-engineered lymphocyte avidity by stabilizing and increasing the expression of the exogenous TCR.…”

Section: Discussionmentioning

confidence: 99%

“…Recently, this approach has been proved clinically beneficial as the transfer of such "reprogrammed" cells to terminally ill cancer patients led to tumor regression in 13% (7) to up to 60% (8) of the patients treated. Although promising, the clinical use of this approach has been hampered by several hurdles, including the proper expression of the exogenous TCR (9,10). In that regard, we and others have devised strategies to increase T cell avidity by modifying essentially TCR extracellular regions, which include the promotion of preferential TCR chain pairing (11)(12)(13)(14)(15)(16), the use of TCR-CD3zeta fusion protein (17), and the deletion of TCR glycosylation sites (18).…”

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confidence: 99%

Incorporation of Transmembrane Hydrophobic Mutations in the TCR Enhance Its Surface Expression and T Cell Functional Avidity

Haga-Friedman

Horovitz-Fried

Cohen

2012

The Journal of Immunology

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TCR-gene transfer represents an effective way to redirect the specificity of T lymphocytes for therapeutic purposes. Recent successful clinical trials have underscored the potential of this approach in which efficient expression of the exogenous TCR has been directly linked to the efficacy of T cell activity. It has been also demonstrated that the TCR exhibits a lack of stability associated with the presence of positively charged residues in its transmembrane (TM) region. In this study, we designed an original approach selectively to improve exogenous TCR stability by increasing the hydrophobic nature of the TCRα TM region. Incorporation of hydrophobic residues at evolutionarily permissive positions resulted in an enhanced surface expression of the TCR chains, leading to an improved cellular avidity and anti-tumor TCR activity. Furthermore, this strategy was successfully applied to different TCRs, enabling the targeting of human tumors from different histologies. We also show that the combination of these hydrophobic mutations with another TCR-enhancing approach further improved TCR expression and function. Overall, these findings provide information regarding TCR TM composition that can be applied for the improvement of TCR-gene transfer-based treatments.

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“…While codon optimization, murinization of human TCRs, and the addition of cysteine residues have been found to reduce mispairing [54], complete knockout of the endogenous TCR would be advantageous. Further work exploring genome editing to knockout endogenous TCRα/β chains has been achieved using ZFNs [55, 56], TALENs [57, 58], megaTAL, and CRISPR/Cas [59] nucleases.…”

Section: Applicationsmentioning

confidence: 99%

Genome-Edited T Cell Therapies

Delhove

Qasim

2017

Curr Stem Cell Rep

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Purpose of ReviewAlternative approaches to conventional drug-based cancer treatments have seen T cell therapies deployed more widely over the last decade. This is largely due to their ability to target and kill specific cell types based on receptor recognition. Introduction of recombinant T cell receptors (TCRs) using viral vectors and HLA-independent T cell therapies using chimeric antigen receptors (CARs) are discussed. This article reviews the tools used for genome editing, with particular emphasis on the applications of site-specific DNA nuclease mediated editing for T cell therapies.Recent FindingsGenetic engineering of T cells using TCRs and CARs with redirected antigen-targeting specificity has resulted in clinical success of several immunotherapies. In conjunction, the application of genome editing technologies has resulted in the generation of HLA-independent universal T cells for allogeneic transplantation, improved T cell sustainability through knockout of the checkpoint inhibitor, programmed cell death protein-1 (PD-1), and has shown efficacy as an antiviral therapy through direct targeting of viral genomic sequences and entry receptors.SummaryThe combined use of engineered antigen-targeting moieties and innovative genome editing technologies have recently shown success in a small number of clinical trials targeting HIV and hematological malignancies and are now being incorporated into existing strategies for other immunotherapies.

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T cell receptor gene therapy: strategies for optimizing transgenic TCR pairing

Cited by 92 publications

References 84 publications

Targeting melanoma with immunoliposomes coupled to anti-MAGE A1 TCR-like single-chain antibody

Targeting melanoma with immunoliposomes coupled to anti-MAGE A1 TCR-like single-chain antibody

Incorporation of Transmembrane Hydrophobic Mutations in the TCR Enhance Its Surface Expression and T Cell Functional Avidity

Genome-Edited T Cell Therapies

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