Increased antitumor efficacy of PD-1-deficient melanoma-specific human lymphocytes

Background Genome editing offers unique perspectives for optimizing the functional properties of T cells for adoptive cell transfer purposes. So far, PDCD1 editing has been successfully tested mainly in chimeric antigen receptor T (CAR-T) cells and human primary T cells. Nonetheless, for patients wi...

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Main Authors: Nathalie Labarrière, Lucine Marotte, Sylvain Simon, Virginie Vignard, Emilie Dupre, Malika Gantier, Jonathan Cruard, Jean-Baptiste Alberge, Melanie Hussong, Cecile Deleine, Jean-Marie Heslan, Jonathan Shaffer, Tiffany Beauvais, Joelle Gaschet, Emmanuel Scotet, Delphine Fradin, Anne Jarry, Tuan Nguyen
Format: Article
Language:English
Published: BMJ Publishing Group 2020-05-01
Series:Journal for ImmunoTherapy of Cancer
Online Access:https://jitc.bmj.com/content/8/1/e000311.full
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author Nathalie Labarrière
Lucine Marotte
Sylvain Simon
Virginie Vignard
Emilie Dupre
Malika Gantier
Jonathan Cruard
Jean-Baptiste Alberge
Melanie Hussong
Cecile Deleine
Jean-Marie Heslan
Jonathan Shaffer
Tiffany Beauvais
Joelle Gaschet
Emmanuel Scotet
Delphine Fradin
Anne Jarry
Tuan Nguyen
author_facet Nathalie Labarrière
Lucine Marotte
Sylvain Simon
Virginie Vignard
Emilie Dupre
Malika Gantier
Jonathan Cruard
Jean-Baptiste Alberge
Melanie Hussong
Cecile Deleine
Jean-Marie Heslan
Jonathan Shaffer
Tiffany Beauvais
Joelle Gaschet
Emmanuel Scotet
Delphine Fradin
Anne Jarry
Tuan Nguyen
author_sort Nathalie Labarrière
collection DOAJ
description Background Genome editing offers unique perspectives for optimizing the functional properties of T cells for adoptive cell transfer purposes. So far, PDCD1 editing has been successfully tested mainly in chimeric antigen receptor T (CAR-T) cells and human primary T cells. Nonetheless, for patients with solid tumors, the adoptive transfer of effector memory T cells specific for tumor antigens remains a relevant option, and the use of high avidity T cells deficient for programmed cell death-1 (PD-1) expression is susceptible to improve the therapeutic benefit of these treatments.Methods Here we used the transfection of CAS9/sgRNA ribonucleoproteic complexes to edit PDCD1 gene in human effector memory CD8+ T cells specific for the melanoma antigen Melan-A. We cloned edited T cell populations and validated PDCD1 editing through sequencing and cytometry in each T cell clone, together with T-cell receptor (TCR) chain’s sequencing. We also performed whole transcriptomic analyses on wild-type (WT) and edited T cell clones. Finally, we documented in vitro and in vivo through adoptive transfer in NOD scid gamma (NSG) mice, the antitumor properties of WT and PD-1KO T cell clones, expressing the same TCR.Results Here we demonstrated the feasibility to edit PDCD1 gene in human effector memory melanoma-specific T lymphocytes. We showed that PD-1 expression was dramatically reduced or totally absent on PDCD1-edited T cell clones. Extensive characterization of a panel of T cell clones expressing the same TCR and exhibiting similar functional avidity demonstrated superior antitumor reactivity against a PD-L1 expressing melanoma cell line. Transcriptomic analysis revealed a downregulation of genes involved in proliferation and DNA replication in PD-1-deficient T cell clones, whereas genes involved in metabolism and cell signaling were upregulated. Finally, we documented the superior ability of PD-1-deficient T cells to significantly delay the growth of a PD-L1 expressing human melanoma tumor in an NSG mouse model.Conclusion The use of such lymphocytes for adoptive cell transfer purposes, associated with other approaches modulating the tumor microenvironment, would be a promising alternative to improve immunotherapy efficacy in solid tumors.
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spelling doaj-art-33f76b9d6e7146feb568dd7170c5ed8c2024-11-09T09:40:08ZengBMJ Publishing GroupJournal for ImmunoTherapy of Cancer2051-14262020-05-018110.1136/jitc-2019-000311Increased antitumor efficacy of PD-1-deficient melanoma-specific human lymphocytesNathalie Labarrière0Lucine Marotte1Sylvain Simon2Virginie Vignard3Emilie Dupre4Malika Gantier5Jonathan Cruard6Jean-Baptiste Alberge7Melanie Hussong8Cecile Deleine9Jean-Marie Heslan10Jonathan Shaffer11Tiffany Beauvais12Joelle Gaschet13Emmanuel Scotet14Delphine Fradin15Anne Jarry16Tuan Nguyen172 LabEx IGO, Université de Nantes, Nantes, France1 Université de Nantes, Inserm, CRCINA, F-44000 Nantes, FranceImmunotherapy Integrated Research Center, Fred Hutchinson Cancer Center, Seattle, Washington, USA1 Université de Nantes, Inserm, CRCINA, F-44000 Nantes, Francede Duve Institute, Université catholique de Louvain, Brussels, Belgium2 LabEx IGO, Université de Nantes, Nantes, France2 LabEx IGO, Université de Nantes, Nantes, France1 Université de Nantes, Inserm, CRCINA, F-44000 Nantes, France4 NGS Assay Research & Development, Qiagen Sciences, Frederick, Maryland, United States1 Université de Nantes, Inserm, CRCINA, F-44000 Nantes, France2 LabEx IGO, Université de Nantes, Nantes, FranceSociology, University of Vermont, Burlington, Vermont, USA1 Inserm UMR1232, CRCINA, Nantes, Pays de la Loire, France1 Université de Nantes, Inserm, CRCINA, F-44000 Nantes, France1 Université de Nantes, Inserm, CRCINA, F-44000 Nantes, France1 Université de Nantes, Inserm, CRCINA, F-44000 Nantes, France1 Université de Nantes, Inserm, CRCINA, F-44000 Nantes, FranceTheravance Biopharma Inc, South San Francisco, California, USABackground Genome editing offers unique perspectives for optimizing the functional properties of T cells for adoptive cell transfer purposes. So far, PDCD1 editing has been successfully tested mainly in chimeric antigen receptor T (CAR-T) cells and human primary T cells. Nonetheless, for patients with solid tumors, the adoptive transfer of effector memory T cells specific for tumor antigens remains a relevant option, and the use of high avidity T cells deficient for programmed cell death-1 (PD-1) expression is susceptible to improve the therapeutic benefit of these treatments.Methods Here we used the transfection of CAS9/sgRNA ribonucleoproteic complexes to edit PDCD1 gene in human effector memory CD8+ T cells specific for the melanoma antigen Melan-A. We cloned edited T cell populations and validated PDCD1 editing through sequencing and cytometry in each T cell clone, together with T-cell receptor (TCR) chain’s sequencing. We also performed whole transcriptomic analyses on wild-type (WT) and edited T cell clones. Finally, we documented in vitro and in vivo through adoptive transfer in NOD scid gamma (NSG) mice, the antitumor properties of WT and PD-1KO T cell clones, expressing the same TCR.Results Here we demonstrated the feasibility to edit PDCD1 gene in human effector memory melanoma-specific T lymphocytes. We showed that PD-1 expression was dramatically reduced or totally absent on PDCD1-edited T cell clones. Extensive characterization of a panel of T cell clones expressing the same TCR and exhibiting similar functional avidity demonstrated superior antitumor reactivity against a PD-L1 expressing melanoma cell line. Transcriptomic analysis revealed a downregulation of genes involved in proliferation and DNA replication in PD-1-deficient T cell clones, whereas genes involved in metabolism and cell signaling were upregulated. Finally, we documented the superior ability of PD-1-deficient T cells to significantly delay the growth of a PD-L1 expressing human melanoma tumor in an NSG mouse model.Conclusion The use of such lymphocytes for adoptive cell transfer purposes, associated with other approaches modulating the tumor microenvironment, would be a promising alternative to improve immunotherapy efficacy in solid tumors.https://jitc.bmj.com/content/8/1/e000311.full
spellingShingle Nathalie Labarrière
Lucine Marotte
Sylvain Simon
Virginie Vignard
Emilie Dupre
Malika Gantier
Jonathan Cruard
Jean-Baptiste Alberge
Melanie Hussong
Cecile Deleine
Jean-Marie Heslan
Jonathan Shaffer
Tiffany Beauvais
Joelle Gaschet
Emmanuel Scotet
Delphine Fradin
Anne Jarry
Tuan Nguyen
Increased antitumor efficacy of PD-1-deficient melanoma-specific human lymphocytes
Journal for ImmunoTherapy of Cancer
title Increased antitumor efficacy of PD-1-deficient melanoma-specific human lymphocytes
title_full Increased antitumor efficacy of PD-1-deficient melanoma-specific human lymphocytes
title_fullStr Increased antitumor efficacy of PD-1-deficient melanoma-specific human lymphocytes
title_full_unstemmed Increased antitumor efficacy of PD-1-deficient melanoma-specific human lymphocytes
title_short Increased antitumor efficacy of PD-1-deficient melanoma-specific human lymphocytes
title_sort increased antitumor efficacy of pd 1 deficient melanoma specific human lymphocytes
url https://jitc.bmj.com/content/8/1/e000311.full
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