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posted on 2024-02-28, 14:20 authored by Tsima Abou Kors, Linda Hofmann, Annika Betzler, Kathrina Payer, Martin Bens, Jens Truong, Adrian von Witzleben, Jaya Thomas, Johann M. Kraus, Randa Kalaajieh, Diana Huber, Jasmin Ezić, Julian Benckendorff, Jens Greve, Patrick J. Schuler, Christian H. Ottensmeier, Hans A. Kestler, Thomas K. Hoffmann, Marie-Nicole Theodoraki, Cornelia Brunner, Simon Laban INHBA expression in OPSCC tumor and cell lines and its correlation with EMT transcription factors. Correlation of INHBA expression with the expression of SNAI1 (A), SNAI2 (B), and TWIST1 (C) in OPSCC tumors (Ulm cohort). TPM = transcripts per million. D, Relative expression of INHBA mRNA in native UDSCC1 (HPV-negative) and UDSCC2 (HPV-positive) cells as determined by qRT-PCR and normalized to RPL30 and UDSCC1. Results are shown as mean ± SD of n = 6 replicates. INHBA expression was not detected in n = 3 UDSCC2 samples. Expression level between UDSCC1 and UDSCC2 was compared by unpaired t test with **, P ≤ 0.01. E, Representative Western blot analysis (of n = 3 replicates) showing INHBA protein expression in native UDSCC1 and UDSCC2 cells. Numbers below blots indicate relative INHBA intensity normalized to GAPDH. F, Relative expression of INHBA, SNAI1, SNAI2, and TWIST1 mRNA in UDSCC1 cells upon transfection with Mock siRNA [Mock knockdown (KD)] and two INHBA siRNAs (INHBA KD1 and KD2) after 24 hours (t24) and 48 hours (t48). Target gene expression was determined by qRT-PCR and normalized to RPL30 and Mock KD. Results are shown as mean ± SD of n = 3 replicates. Expression level between Mock KD and INHBA KD1/2 was compared by unpaired t test with *, P ≤ 0.05; **, P ≤ 0.01; and ns, P > 0.05. G, Representative Western blot analysis (of n = 3 replicates) showing INHBA protein expression in Mock KD and INHBA KD1/KD2 at t24 and t48. Numbers below blots indicate relative INHBA intensity normalized to GAPDH.
Funding
Deutsche Forschungsgemeinschaft (DFG)
History
ARTICLE ABSTRACT
Patients with oropharyngeal squamous cell carcinoma (OPSCC) caused by human papilloma virus (HPV) exhibit a better prognosis than those with HPV-negative OPSCC. This study investigated the distinct molecular pathways that delineate HPV-negative from HPV-positive OPSCC to identify biologically relevant therapeutic targets. Bulk mRNA from 23 HPV-negative and 39 HPV-positive OPSCC tumors (n = 62) was sequenced to uncover the transcriptomic profiles. Differential expression followed by gene set enrichment analysis was performed to outline the top enriched biological process in the HPV-negative compared with HPV-positive entity. INHBA, the highest overexpressed gene in the HPV-negative tumor, was knocked down. Functional assays (migration, proliferation, cell death, stemness) were conducted to confirm the target's oncogenic role. Correlation analyses to reveal its impact on the tumor microenvironment were performed. We revealed that epithelial-to-mesenchymal transition (EMT) is the most enriched process in HPV-negative compared with HPV-positive OPSCC, with INHBA (inhibin beta A subunit) being the top upregulated gene. INHBA knockdown downregulated the expression of EMT transcription factors and attenuated migration, proliferation, stemness, and cell death resistance of OPSCC cells. We uncovered that INHBA associates with a pro-tumor microenvironment by negatively correlating with antitumor CD8+ T and B cells while positively correlating with pro-tumor M1 macrophages. We identified three miRNAs that are putatively involved in repressing INHBA expression. Our results indicate that the upregulation of INHBA is tumor-promoting. We propose INHBA as an attractive therapeutic target for the treatment of INHBA-enriched tumors in patients with HPV-negative OPSCC to ameliorate prognosis.
Patients with HPV-negative OPSCC have a poorer prognosis due to distinct molecular pathways. This study reveals significant transcriptomic differences between HPV-negative and HPV-positive OPSCC, identifying INHBA as a key upregulated gene in HPV-negative OPSCC's oncogenic pathways. INHBA is crucial in promoting EMT, cell proliferation, and an immunosuppressive tumor environment, suggesting its potential as a therapeutic target for HPV-negative OPSCC.
