High-avidity TCR signaling drives an NKG2A-positive CD8 T cell exhaustion state in human tumors
Year: 2026
Type: preprint
Abstract: ABSTRACT T cells adopt diverse states in human disease, with tumor-specific CD8 T cells serving as the quintessential example. These cells are critical for immunotherapy response, yet acquire hypofunctional states of exhaustion that impair treatment efficacy. While significant heterogeneity is observed among exhausted CD8 T cells (T EX ), the mechanisms directing their unique phenotypes remain unresolved, limiting our efforts to augment T EX cell function. Here, we profile T EX cells in human head and neck squamous cell carcinoma (HNSCC) tumors and define a major mechanism regulating their cell state. With single-cell RNA and T cell receptor (TCR) sequencing of 106,667 tumor-infiltrating CD8 T cells, we define and validate three fundamental T EX subsets, each harboring dominant clonotypes: a progenitor subset (T PEX ) and two intra -tumoral populations marked by intermediate (T EX-int ) or high expression of inhibitory receptors, including killer cell lectin-like receptors, such as KLRC1 /NKG2A (T EX-KLR ). Using an in vitro model, we show that TCR avidity-dependent NFAT signaling governs an axis between the T EX-int and T EX-KLR states across human cancers, with high-avidity signaling driving the T EX-KLR state. Additionally, we uncover that interleukin-12 signaling selectively antagonizes high-avidity T EX-KLR -specific genes, despite broadly potentiating activation and cytotoxicity. Finally, we find that T EX-int and T EX-KLR cells are associated with anti-PD-1 response in HNSCC, with both of these cell types enriched near viable cancer cells following treatment. Together, these findings reveal that TCR avidity orchestrates tumor-infiltrating T EX cell states through NFAT signaling – a tunable axis that may be targeted to improve T EX cell function.
Authors: Riley D.Z. Mullins, Jesse M. Zaretsky, Thomas F. Barrett, Emily Stoller, Porter Bischoff, Ann Marie Egloff, Cody Huffman, Salma Ramadan, Amna Ramadan, Jessica Ley, Katherine Schwetye, Daniel Miller, Jennifer De Los Santos, Anthony J. Apicelli, Nikhil Rammohan, Wade Thorstad, Peter Oppelt, Brendan J. Knapp, Christine Auberle, Guangyong Peng, Craig Buchman, Andres Bur, Ben Wahle, Jason T. Rich, Ryan S. Jackson, Patrik Pipkorn, Paul A. Zolkind, Richard A. Harbison, Takeshi Egawa, Nathan Singh, Robi D. Mitra, Sana D. Karam, Douglas R. Adkins, Ravindra Uppaluri, Sidharth V. Puram
Language: English
Subfield: Oncology
Field: Medicine
Domain: Health Sciences
Open Access status: green
Awards: Understanding programs of invasion and metastasis in head and neck cancer, RNA modification and the regulation of partial EMT in head and neck cancer, The Determinants of CD8 T cell Dysfunction in Oral Cavity Squamous Cell Carcinoma, Deciphering epigenetically-regulated pathways to improve targeted therapy for invasion and metastasis in head and neck cancer, TRAINING IN THE IMMUNOBIOLOGY AND MOLECULAR CELL BIOLOGY OF CANCER, The transcription factor c-MYC in lymphocyte expansion and restriction of stemness, Molecular mechanisms of CD8 T cell fate decision instructed by cytokine signaling, Washington University Paul Calabresi K12 Career Development Award for Clinical Oncology, Dissecting hybrid epithelial-mesenchymal states in head and neck cancer
OpenAlex ID: W7163395623