Three-dimensional collagen-based scaffold model to study the microenvironment and drug-resistance mechanisms of oropharyngeal squamous cell carcinomas

Objective:Squamous cell carcinoma(SCC)represents the most common histotype of all head and neck malignancies and includes oropharyngeal squamous cell carcinoma(OSCC),a tumor associated with different clinical outcomes and linked to human papilloma virus(HPV)status.Translational research has few available in vitro models with which to study the different pathophysiological behavior of OSCCs.The present study proposes a 3-dimensional(3 D)biomimetic collagen-based scaffold to mimic the tumor microenvironment and the crosstalk between the extracellular matrix(ECM)and cancer cells.Methods:We compared the phenotypic and genetic features of HPV-positive and HPV-negative OSCC cell lines cultured on common monolayer supports and on scaffolds.We also explored cancer cell adaptation to the 3 D microenvironment and its impact on the efficacy of drugs tested on cell lines and primary cultures.Results:HPV-positive and HPV-negative cell lines were successfully grown in the 3 D model and displayed different collagen fiber organization.The 3 D cultures induced an increased expression of markers related to epithelial–mesenchymal transition(EMT)and to matrix interactions and showed different migration behavior,as confirmed by zebrafish embryo xenografts.The expression of hypoxia-inducible factor 1α(1α)and glycolysis markers were indicative of the development of a hypoxic microenvironment inside the scaffold area.Furthermore,the 3 D cultures activated drug-resistance signaling pathways in both cell lines and primary cultures.Conclusions:Our results suggest that collagen-based scaffolds could be a suitable model for the reproduction of the pathophysiological features of OSCCs.Moreover,3 D architecture appears capable of inducing drug-resistance processes that can be studied to better our understanding of the different clinical outcomes of HPV-positive and HPV-negative patients with OSCCs.

Therapeutic Outcomes of HPV Positive and HPV Negative Oropharyngeal Squamous Cell Carcinomas

Background & Objectives: HPV associated (HPV+) oropharyngeal cancers, compared with HPV non-associated ones (HPV?) have different characters and significantly better outcomes. HPV and cancer characteristics differ among countries and few data is available in Indian population regarding this issue. Thus, we attempted to determine the treatment response and survival rates between HPV+ versus HPV negative oropharyngeal cancers in Indian population. Methods: This prospective observational study was conducted from December 2016 - October 2018 in patients with stage III/IV oropharyngeal squamous cell carcinomas (SCC) (n = 65). The patients received induction chemotherapy with Docetaxel, Cisplatin, 5-Fluorouracil × 3 cycles, followed by concurrent chemoradiation 66 Gy as 33 fractions along with cisplatin 40 mg/m2 weekly. The primary outcome was treatment response rate, which was defined as per Response Evaluation Criteria in Solid Tumours (RECIST) criteria. Secondary outcomes were clinico-pathological differences between two groups, overall survival (OS), progression free survival (PFS). Results: Of the 65 patients included in the study, 17 were HPV positive and 48 patients were HPV negative. Median age in HPV positive arm is 48 years and HPV negative arm is 59 years. HPV positive patients presented with early T stage and advanced nodal (N) stage. Most common histopathology in both arms was moderately differentiated squamous cell carcinomas followed by well differentiated squamous cell carcinomas. HPV+ vs HPV? showed the following, treatment response 82.4% vs 52.1% (p-0.029). 1 year progression free survival (PFS) of 76.5% vs 52.1% in HPV negative arm (p-0.08) & 1 year overall survival was 82.4% vs 70.8% (p-0.353). Grade 3 or 4 toxicities did not differ significantly between HPV positive and HPV negative arms. Conclusion: HPV positive oropharyngeal SCC patients showed significantly better treatment response than HPV negative ones. Progression free survival, overall survival and toxicity profile did not differ significantly between the two groups. Although due to small size, we did not compare data stratified by the cancer characteristics, the data is worthy to further characterize this tumour especially as HPV positive versus HPV negative.

Detection of human papillomavirus in oropharyngeal squamous cell carcinoma

Worldwide there has been a significant increase in the incidence of oropharyngeal squamous cell carcinoma(OPSCC)etiologically attributed to oncogenic human papillomavirus(HPV).Reliable and accurate identification and detection tools are important as the incidence of HPV-related cancer is on the rise.Several HPV detection methods for OPSCC have been developed and each has its own advantages and disadvantages in regard to sensitivity,specificity,and technical difficulty.This review summarizes our current knowledge of molecular methods for detecting HPV in OPSCC,including HPV DNA/RNA polymerase chain reaction(PCR),loop-mediated isothermal amplification(LAMP),p16 immunohistochemistry(IHC),and DNA/RNA in situ hybridization(ISH)assays.This summary may facilitate the selection of a suitable method for detecting HPV infection,and therefore may help in the early diagnosis of HPV-related carcinoma to reduce its mortality,incidence,and morbidity.