MRNA and miRNA Profiles of Exosomes from Cultured Tumor Cells Reveal Biomarkers Specific for HPV16-Positive and HPV16-Negative Head and Neck Cancer.

Sonja Ludwig,Muller Fabbri,Petra Wise,Janette Lamb,Richard Sposto,Priyanka Sharma,Deborah Hollingshead,Stephan Lang,Theresa L Whiteside

doi:10.3390/ijms21228570

Abstract

Human papillomavirus (HPV)(+) and HPV(−) head and neck cancer (HNC) cells’ interactions with the host immune system are poorly understood. Recently, we identified molecular and functional differences in exosomes produced by HPV(+) vs. HPV(−) cells, suggesting that genetic cargos of exosomes might identify novel biomarkers in HPV-related HNCs. Exosomes were isolated by size exclusion chromatography from supernatants of three HPV(+) and two HPV(−) HNC cell lines. Paired cell lysates and exosomes were analyzed for messenger RNA (mRNA) by qRT-PCR and microRNA (miR) contents by nanostring analysis. The mRNA profiles of HPV(+) vs. HPV(−) cells were distinct, with EGFR, TP53 and HSPA1A/B overexpressed in HPV(+) cells and IL6, FAS and DPP4 in HPV(−) cells. The mRNA profiles of HPV(+) or HPV(−) exosomes resembled the cargo of their parent cells. miR expression profiles in cell lysates identified 8 miRs expressed in HPV(−) cells vs. 14 miRs in HPV(+) cells. miR-205-5p was exclusively expressed in HPV(+) exosomes, and miR-1972 was only detected in HPV(−) exosomes. We showed that HPV(+) and HPV(−) exosomes recapitulated the mRNA expression profiles of their parent cells. Expression of miRs was dependent on the HPV status, and miR-205-5p in HPV(+) and miR-1972 in HPV(−) exosomes emerge as potential discriminating HPV-associated biomarkers.

Highlights

Head and neck cancers (HNCs) belong to the ten most common malignant diseases worldwide [1]
We report that the messenger RNA (mRNA) profiles of HNC cells and exosomes these cells produced were similar, human papillomavirus (HPV)(+) cells and exosomes were enriched in HPV-related transcripts
These exosomes co-incubated with human primary T cells suppressed T-cell activation and proliferation and induced T-cell apoptosis regardless of the HPV status of parent cells [17]

Summary

Introduction

Head and neck cancers (HNCs) belong to the ten most common malignant diseases worldwide [1]. Due to their distinct pathogenesis, HNCs are heterogeneous, and symptoms often appear when tumors are in late stages. The HPV(+) oropharyngeal squamous cell carcinoma (OPSCC) differs from other HNCs by histopathologic, molecular and clinical characteristics. It is found in younger patients who usually do not have a history of alcohol abuse or smoking [5]. At the time of diagnosis, the majority of HNCs are locally advanced, and the standard therapy regimens are given to patients regardless of their HPV status. The use of more targeted and less toxic therapies, e.g., immunotherapies in combination with chemotherapy, for treatment of HPV(+) patients offers an opportunity for improvement [9,10]

Methods

Results

Conclusion