Genetic Mutation and Exosome Signature of Human Papilloma Virus Associated Oropharyngeal Cancer

Abstract

Human papilloma virus-16 (HPV-16) associated oropharyngeal cancer (HPVOPC) is increasing alarmingly in the United States. We performed whole genome sequencing of a 44 year old, male HPVOPC subject diagnosed with moderately differentiated tonsillar carcinoma. We identified new somatic mutation in MUC16 (A.k.a. CA-125), MUC12, MUC4, MUC6, MUC2, SIRPA, HLA-DRB1, HLA-A and HLA-B molecules. Increased protein expression of MUC16, SIRPA and decreased expression of HLA-DRB1 was further demonstrated in this HPVOPC subject and an additional set of 15 HPVOPC cases. Copy number gain (3 copies) was also observed for MUC2, MUC4, MUC6 and SIRPA. Enhanced expression of MUC16, SIRPA and HPV-16-E7 protein was detectable in the circulating exosomes of numerous HPVOPC subjects. Treatment of non-tumorigenic mammary epithelial cells with exosomes derived from aggressive HPVOPC cells harboring MUC16, SIRPA and HPV-16-E7 proteins augmented invasion and induced epithelial to mesenchymal transition (EMT) accompanied by an increased expression ratio of the EMT markers Vimentin/E-cadherin. Exosome based screening of key HPVOPC associated molecules could be beneficial for early cancer diagnosis, monitoring and surveillance.

Expression of human papillomavirus oncoproteins E6 and E7 inhibits invadopodia activity but promotes cell migration in HPV‐positive head and neck squamous cell carcinoma cells

“…HPV − SCC‐25, SCC‐61, and SCC‐1 cells are frequently used as cell line models for studying invadopodia . HPV + SCC‐47 and SCC‐104 cells are common cell line models for studying the effects of HPV‐16 . We performed invadopodia assays on these cell lines and quantitated invadopodia numbers and ECM degradation of a fluorescently labeled ECM (Figure B‐D).…”

Human Papillomavirus and carcinogenesis: Novel mechanisms of cell communication involving extracellular vesicles

“…In addition, the recurrence of HPV + oropharyngeal cancer manifests later than in HPV − patients [38,[42][43][44]. Using whole genome sequencing analysis of HPV + oropharyngeal cancer specimen, it has been identified a panel of mutated genes involved in malignant progression including several members of Mucin family, HLA-A, -B and DRB1 as well as the surface marker CD172a/SIRPA [45]. These mutations are responsible for the overexpression of MUC16 and SIRPA in HPV + oropharyngeal cancer as well as in circulating exosomes isolated from blood of patients.…”

“…These mutations are responsible for the overexpression of MUC16 and SIRPA in HPV + oropharyngeal cancer as well as in circulating exosomes isolated from blood of patients. EVs derived from HPV + oropharyngeal cancer specimen possess the ability to induce both epithelial to mesenchymal transition (EMT) and increase the migration and invasion of a HPV − mammary epithelial cell line and, contrary to EVs derived from anogenital squamous cell carcinoma, they express HPV16 E7 [45].…”

Role of Extracellular Vesicles in Human Papillomavirus-Induced Tumorigenesis

“…(HPVOPC), serum collected from patients contains Exos expressing key altered proteins, namely, mucin-16 (MUC-16) and signal regulatory protein α (SIRPA) as well as E7 oncoprotein [81]. Similarly, Exos derived from a HPVOPC cell line displayed the ability to induce epithelial-mesenchymal transition (EMT) and invasiveness of two human mammary epithelial cell lines [81]. Finally, the presence of EVs containing HPV DNA was analyzed in the serum of patients with breast cancer.…”