TMED inhibition suppresses cell surface PD-1 expression and overcomes T cell dysfunction

Background Blockade of the programmed cell death protein 1 (PD-1) immune checkpoint (ICB) is revolutionizing cancer therapy, but little is known about the mechanisms governing its expression on CD8 T cells. Because PD-1 is induced during activation of T cells, we set out to uncover regulators whose...

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Main Authors: Gal Markel, Michal J Besser, Eytan Ruppin, Oscar Krijgsman, Daniel S Peeper, Judit Díaz-Gómez, Ettai Markovits, David W Vredevoogd, Georgi Apriamashvili, Pierre L Levy, Sanju Sinha, Zowi R Huinen, Nils L Visser, Beaunelle de Bruijn, Julia Boshuizen, Susan E van Hal-van Veen, Maarten A Ligtenberg, Onno B Bleijerveld, Chun-Pu Lin, Santiago Duro Sánchez, Juan Simon Nieto, Alex van Vliet, Maarten Altelaar
Format: Article
Language:English
Published: BMJ Publishing Group 2024-11-01
Series:Journal for ImmunoTherapy of Cancer
Online Access:https://jitc.bmj.com/content/12/11/e010145.full
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author Gal Markel
Michal J Besser
Eytan Ruppin
Oscar Krijgsman
Daniel S Peeper
Judit Díaz-Gómez
Ettai Markovits
David W Vredevoogd
Georgi Apriamashvili
Pierre L Levy
Sanju Sinha
Zowi R Huinen
Nils L Visser
Beaunelle de Bruijn
Julia Boshuizen
Susan E van Hal-van Veen
Maarten A Ligtenberg
Onno B Bleijerveld
Chun-Pu Lin
Santiago Duro Sánchez
Juan Simon Nieto
Alex van Vliet
Maarten Altelaar
author_facet Gal Markel
Michal J Besser
Eytan Ruppin
Oscar Krijgsman
Daniel S Peeper
Judit Díaz-Gómez
Ettai Markovits
David W Vredevoogd
Georgi Apriamashvili
Pierre L Levy
Sanju Sinha
Zowi R Huinen
Nils L Visser
Beaunelle de Bruijn
Julia Boshuizen
Susan E van Hal-van Veen
Maarten A Ligtenberg
Onno B Bleijerveld
Chun-Pu Lin
Santiago Duro Sánchez
Juan Simon Nieto
Alex van Vliet
Maarten Altelaar
author_sort Gal Markel
collection DOAJ
description Background Blockade of the programmed cell death protein 1 (PD-1) immune checkpoint (ICB) is revolutionizing cancer therapy, but little is known about the mechanisms governing its expression on CD8 T cells. Because PD-1 is induced during activation of T cells, we set out to uncover regulators whose inhibition suppresses PD-1 abundance without adversely impacting on T cell activation.Methods To identify PD-1 regulators in an unbiased fashion, we performed a whole-genome, fluorescence-activated cell sorting (FACS)-based CRISPR-Cas9 screen in primary murine CD8 T cells. A dual-readout design using the activation marker CD137 allowed us to uncouple genes involved in PD-1 regulation from those governing general T cell activation.Results We found that the inactivation of one of several members of the TMED/EMP24/GP25L/p24 family of transport proteins, most prominently TMED10, reduced PD-1 cell surface abundance, thereby augmenting T cell activity. Another client protein was cytotoxic T lymphocyte-associated protein 4 (CTLA-4), which was also suppressed by TMED inactivation. Treatment with TMED inhibitor AGN192403 led to lysosomal degradation of the TMED-PD-1 complex and reduced PD-1 abundance in tumor-infiltrating CD8 T cells (TIL) in mice, thus reversing T cell dysfunction. Clinically corroborating these findings, single-cell RNA analyses revealed a positive correlation between TMED expression in CD8 TIL, and both a T cell dysfunction signature and lack of ICB response. Similarly, patients receiving a TIL product with high TMED expression had a shorter overall survival.Conclusion Our results uncover a novel mechanism of PD-1 regulation, and identify a pharmacologically tractable target whose inhibition suppresses PD-1 abundance and T cell dysfunction.
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spelling doaj-art-b3b1db25d0594fcdad82ad2a293ad0502025-08-20T03:54:01ZengBMJ Publishing GroupJournal for ImmunoTherapy of Cancer2051-14262024-11-01121110.1136/jitc-2024-010145TMED inhibition suppresses cell surface PD-1 expression and overcomes T cell dysfunctionGal Markel0Michal J Besser1Eytan Ruppin2Oscar Krijgsman3Daniel S Peeper4Judit Díaz-Gómez5Ettai Markovits6David W Vredevoogd7Georgi Apriamashvili8Pierre L Levy9Sanju Sinha10Zowi R Huinen11Nils L Visser12Beaunelle de Bruijn13Julia Boshuizen14Susan E van Hal-van Veen15Maarten A Ligtenberg16Onno B Bleijerveld17Chun-Pu Lin18Santiago Duro Sánchez19Juan Simon Nieto20Alex van Vliet21Maarten Altelaar22Davidoff Center and Samueli Integrative Cancer Pioneering Center, Rabin Medical Center, Petah Tikva, IsraelDepartment of Clinical Microbiology and Immunology, Faculty of Medical & Health Sciences, Tel Aviv University, Tel Aviv, Israel23 Cancer Data Science Laboratory, National Cancer Institute, Bethesda, Maryland, USADepartment of Molecular oncology and immunology, Netherlands Cancer Institute, Oncode Institute, Amsterdam, The NetherlandsDepartment of Molecular oncology and immunology, Netherlands Cancer Institute, Oncode Institute, Amsterdam, The NetherlandsDepartment of Molecular oncology and immunology, Netherlands Cancer Institute, Oncode Institute, Amsterdam, The NetherlandsNucleai, Tel Aviv, IsraelDepartment of Molecular oncology and immunology, Netherlands Cancer Institute, Oncode Institute, Amsterdam, The NetherlandsDepartment of Molecular oncology and immunology, Netherlands Cancer Institute, Oncode Institute, Amsterdam, The NetherlandsDepartment of Molecular oncology and immunology, Netherlands Cancer Institute, Oncode Institute, Amsterdam, The NetherlandsCancer Data Science Laboratory, National Cancer Institute Center for Cancer Research, Bethesda, Maryland, USADepartment of Molecular oncology and immunology, Netherlands Cancer Institute, Oncode Institute, Amsterdam, The NetherlandsDepartment of Molecular oncology and immunology, Netherlands Cancer Institute, Oncode Institute, Amsterdam, The NetherlandsDepartment of Molecular oncology and immunology, Netherlands Cancer Institute, Oncode Institute, Amsterdam, The NetherlandsDepartment of Molecular oncology and immunology, Netherlands Cancer Institute, Oncode Institute, Amsterdam, The NetherlandsDepartment of Molecular oncology and immunology, Netherlands Cancer Institute, Oncode Institute, Amsterdam, The NetherlandsDepartment of Molecular oncology and immunology, Netherlands Cancer Institute, Oncode Institute, Amsterdam, The NetherlandsDepartment of Molecular oncology and immunology, Netherlands Cancer Institute, Oncode Institute, Amsterdam, The NetherlandsDepartment of Molecular oncology and immunology, Netherlands Cancer Institute, Oncode Institute, Amsterdam, The NetherlandsDepartment of Molecular oncology and immunology, Netherlands Cancer Institute, Oncode Institute, Amsterdam, The NetherlandsDepartment of Molecular oncology and immunology, Netherlands Cancer Institute, Oncode Institute, Amsterdam, The NetherlandsDepartment of Molecular oncology and immunology, Netherlands Cancer Institute, Oncode Institute, Amsterdam, The NetherlandsDepartment of Molecular oncology and immunology, Netherlands Cancer Institute, Oncode Institute, Amsterdam, The NetherlandsBackground Blockade of the programmed cell death protein 1 (PD-1) immune checkpoint (ICB) is revolutionizing cancer therapy, but little is known about the mechanisms governing its expression on CD8 T cells. Because PD-1 is induced during activation of T cells, we set out to uncover regulators whose inhibition suppresses PD-1 abundance without adversely impacting on T cell activation.Methods To identify PD-1 regulators in an unbiased fashion, we performed a whole-genome, fluorescence-activated cell sorting (FACS)-based CRISPR-Cas9 screen in primary murine CD8 T cells. A dual-readout design using the activation marker CD137 allowed us to uncouple genes involved in PD-1 regulation from those governing general T cell activation.Results We found that the inactivation of one of several members of the TMED/EMP24/GP25L/p24 family of transport proteins, most prominently TMED10, reduced PD-1 cell surface abundance, thereby augmenting T cell activity. Another client protein was cytotoxic T lymphocyte-associated protein 4 (CTLA-4), which was also suppressed by TMED inactivation. Treatment with TMED inhibitor AGN192403 led to lysosomal degradation of the TMED-PD-1 complex and reduced PD-1 abundance in tumor-infiltrating CD8 T cells (TIL) in mice, thus reversing T cell dysfunction. Clinically corroborating these findings, single-cell RNA analyses revealed a positive correlation between TMED expression in CD8 TIL, and both a T cell dysfunction signature and lack of ICB response. Similarly, patients receiving a TIL product with high TMED expression had a shorter overall survival.Conclusion Our results uncover a novel mechanism of PD-1 regulation, and identify a pharmacologically tractable target whose inhibition suppresses PD-1 abundance and T cell dysfunction.https://jitc.bmj.com/content/12/11/e010145.full
spellingShingle Gal Markel
Michal J Besser
Eytan Ruppin
Oscar Krijgsman
Daniel S Peeper
Judit Díaz-Gómez
Ettai Markovits
David W Vredevoogd
Georgi Apriamashvili
Pierre L Levy
Sanju Sinha
Zowi R Huinen
Nils L Visser
Beaunelle de Bruijn
Julia Boshuizen
Susan E van Hal-van Veen
Maarten A Ligtenberg
Onno B Bleijerveld
Chun-Pu Lin
Santiago Duro Sánchez
Juan Simon Nieto
Alex van Vliet
Maarten Altelaar
TMED inhibition suppresses cell surface PD-1 expression and overcomes T cell dysfunction
Journal for ImmunoTherapy of Cancer
title TMED inhibition suppresses cell surface PD-1 expression and overcomes T cell dysfunction
title_full TMED inhibition suppresses cell surface PD-1 expression and overcomes T cell dysfunction
title_fullStr TMED inhibition suppresses cell surface PD-1 expression and overcomes T cell dysfunction
title_full_unstemmed TMED inhibition suppresses cell surface PD-1 expression and overcomes T cell dysfunction
title_short TMED inhibition suppresses cell surface PD-1 expression and overcomes T cell dysfunction
title_sort tmed inhibition suppresses cell surface pd 1 expression and overcomes t cell dysfunction
url https://jitc.bmj.com/content/12/11/e010145.full
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