Integrative analysis reveals a macrophage-predominant, immunosuppressive immune microenvironment and subtype-specific therapeutic vulnerabilities in advanced salivary gland cancer

Author:

Zuljan Erika,von der Emde Benjamin,Piwonski Iris,Afonseca Pestana Ana Cristina,Klinghammer Konrad,Mock Andreas,Horak PeterORCID,Heining Christoph,Klauschen Frederick,Pretzell Ina,Boerries Melanie,Brandts Christian HORCID,Kreutzfeldt SimonORCID,Teleanu Maria-Veronica,Hübschmann DanielORCID,Morris Luc G T,Keller UlrichORCID,Glimm Hanno,Fröhling Stefan,Ochsenreither Sebastian,Keilholz Ulrich,Blanc Eric,Beule Dieter,Rieke Damian TORCID

Abstract

ABSTRACTBackgroundSalivary gland cancers (SGC) are rare and heterogeneous malignant tumors. Advanced SGC lack established treatment options and show poor response to immunotherapy. Here, an integrative multi-omics analysis in a large cohort of advanced SGC revealed insights into the tumor immune microenvironment (TIM) and distinct mechanisms of immune evasion.MethodsA total of 104 patients with recurrent/metastatic SGC from the DKTK MASTER program were included in this study. Whole-exome or whole-genome sequencing and RNA-sequencing was performed on fresh frozen tumor tissue. The tumor immune microenvironment was analyzed using CIBERSORT deconvolution analysis and immune gene expression scores in bulk RNA-sequencing data. Single-nuclei sequencing and immunohistochemistry analyses were performed in selected samples. Results were validated in bulk RNA-sequencing data of a previously published independent dataset.ResultsBulk transcriptome analysis revealed an immune-deserted TIM in the majority of advanced SGC samples. Immune exclusion was most prominent in adenoid cystic carcinoma (ACC) subgroup 1 exhibiting a downregulation of the antigen processing machinery. Only a small subset of advanced SGC, including few adenoid cystic carcinoma, exhibited T-cell inflammation, which was correlated with tumor mutational burden in Non-ACC samples. Subtype specific expression of immune checkpoints as well as cancer testis antigens were identified with prominent expression of VTCN1 in luminal cells within ACC. Single-cell RNA-sequencing and bulk RNA-seq deconvolution analysis validated immune cell exclusion and revealed a TIM that was dominated by M2 macrophages across SGC subtypes. Among evaluable patients treated with immune checkpoint inhibitors, a high T-cell to macrophage ratio was associated with clinical benefit.ConclusionsThese data support biomarker-based development of immune-checkpoint inhibition and the development of novel immune-checkpoint inhibitors and cellular therapies in SGC.Trial RegistrationRetrospectively registered,NCT05852522

Publisher

Cold Spring Harbor Laboratory

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