Frontiers in Immunology (Jul 2024)

Multi-omics analysis of overexpressed tumor-associated proteins: gene expression, immunopeptide presentation, and antibody response in oropharyngeal squamous cell carcinoma, with a focus on cancer-testis antigens

  • Tsima Abou Kors,
  • Matthias Meier,
  • Lena Mühlenbruch,
  • Lena Mühlenbruch,
  • Lena Mühlenbruch,
  • Lena Mühlenbruch,
  • Annika C. Betzler,
  • Annika C. Betzler,
  • Franziska Oliveri,
  • Martin Bens,
  • Jaya Thomas,
  • Johann M. Kraus,
  • Johannes Doescher,
  • Johannes Doescher,
  • Adrian von Witzleben,
  • Linda Hofmann,
  • Jasmin Ezic,
  • Diana Huber,
  • Julian Benckendorff,
  • Thomas F. E. Barth,
  • Jens Greve,
  • Patrick J. Schuler,
  • Patrick J. Schuler,
  • Cornelia Brunner,
  • Cornelia Brunner,
  • Jonathan M. Blackburn,
  • Thomas K. Hoffmann,
  • Thomas K. Hoffmann,
  • Christian Ottensmeier,
  • Hans A. Kestler,
  • Hans A. Kestler,
  • Hans-Georg Rammensee,
  • Hans-Georg Rammensee,
  • Hans-Georg Rammensee,
  • Juliane S. Walz,
  • Juliane S. Walz,
  • Juliane S. Walz,
  • Juliane S. Walz,
  • Simon Laban,
  • Simon Laban

DOI
https://doi.org/10.3389/fimmu.2024.1408173
Journal volume & issue
Vol. 15

Abstract

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IntroductionThe human leukocyte antigen complex (HLA) is essential for inducing specific immune responses to cancer by presenting tumor-associated peptides (TAP) to T cells. Overexpressed tumor associated antigens, mainly cancer-testis antigens (CTA), are outlined as essential targets for immunotherapy in oropharyngeal squamous cell carcinoma (OPSCC). This study assessed the degree to which presentation, gene expression, and antibody response (AR) of TAP, mainly CTA, are correlated in OPSCC patients to evaluate their potential as immunotherapy targets.Materials and methodsSnap-frozen tumor (NLigand/RNA=40), healthy mucosa (NRNA=6), and healthy tonsils (NLigand=5) samples were obtained. RNA-Seq was performed using Illumina HiSeq 2500/NovaSeq 6000 and whole exome sequencing (WES) utilizing NextSeq500. HLA ligands were isolated from tumor tissue using immunoaffinity purification, UHPLC, and analyzed by tandem MS. Antibodies were measured in serum (NAb=27) utilizing the KREX™ CT262 protein array. Data analysis focused on 312 proteins (KREX™ CT262 panel + overexpressed self-proteins).Results183 and 94 of HLA class I and II TAP were identified by comparative profiling with healthy tonsils. Genes from 26 TAP were overexpressed in tumors compared to healthy mucosa (LFC>1; FDR<0.05). Low concordance (r=0.25; p<0.0001) was found between upregulated mRNA and class I TAP. The specific mode of correlation of TAP was found to be dependent on clinical parameters. A lack of correlation was observed both between mRNA and class II TAP, as well as between class II tumor-unique TAP (TAP-U) presentation and antibody response (AR) levels.DiscussionThis study demonstrates that focusing exclusively on gene transcript levels fails to capture the full extent of TAP presentation in OPSCC. Furthermore, our findings reveal that although CTA are presented at relatively low levels, a few CTA TAP-U show potential as targets for immunotherapy.

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