TY - JOUR
T1 - TMED inhibition suppresses cell surface PD-1 expression and overcomes T cell dysfunction
AU - Vredevoogd, David W.
AU - Apriamashvili, Georgi
AU - Levy, Pierre L.
AU - Sinha, Sanju
AU - Huinen, Zowi R.
AU - Visser, Nils L.
AU - de Bruijn, Beaunelle
AU - Boshuizen, Julia
AU - van Hal-van Veen, Susan E.
AU - Ligtenberg, Maarten A.
AU - Bleijerveld, Onno B.
AU - Lin, Chun Pu
AU - Díaz-Gómez, Judit
AU - Sánchez, Santiago Duro
AU - Markovits, Ettai
AU - Simon Nieto, Juan
AU - van Vliet, Alex
AU - Krijgsman, Oscar
AU - Markel, Gal
AU - Besser, Michal J.
AU - Altelaar, Maarten
AU - Ruppin, Eytan
AU - Peeper, Daniel S.
N1 - Publisher Copyright:
© Author(s) (or their employer(s)) 2024. Re-use permitted under CC BY. Published by BMJ.
PY - 2024/11/7
Y1 - 2024/11/7
N2 - 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.
AB - 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.
KW - Adoptive cell therapy - ACT
KW - Immune checkpoint inhibitor
KW - Immunotherapy
KW - T cell
KW - Tumor infiltrating lymphocyte - TIL
UR - http://www.scopus.com/inward/record.url?scp=85209166982&partnerID=8YFLogxK
U2 - 10.1136/jitc-2024-010145
DO - 10.1136/jitc-2024-010145
M3 - Article
C2 - 39510795
AN - SCOPUS:85209166982
SN - 2051-1426
VL - 12
JO - Journal for ImmunoTherapy of Cancer
JF - Journal for ImmunoTherapy of Cancer
IS - 11
M1 - e010145
ER -