A 4-gene signature from histologically normal surgical margins predicts local recurrence in patients with oral carcinoma: clinical validation

Abstract

Prognostic biomarkers for recurrence of Oral Squamous Cell Carcinoma (OSCC) are urgently needed. We aimed to independently validate a 4-gene expression signature (MMP1, COL4A1, P4HA2, THBS2) predictive of OSCC recurrence risk. Gene expression was measured using Nanostring nCounter ® in 245 histologically normal surgical resection margins from 62 patients. Association between risk scores for individual patients and recurrence was assessed by Kaplan-Meier analysis. Signature performance was quantified by concordance index (CI), hazard ratio (HR) and the area under receiver operating characteristics (AUC). Risk scores for recurrence were significantly higher than recurrence-free patients (p = 9.58e-7, Welch's t-test). A solid performance of the 4-gene signature was determined: CI = 0.64, HR = 3.38 (p = 1.4E-4; log-rank test), AUC = 0.71. We showed that three margins per patient are sufficient to preserve predictive performance (CI = 0.65; HR = 2.92; p = 2.94e-3; AUC = 0.71). Association between the predicted risk scores and recurrence was assessed and showed HR = 2.44 (p = 9.6E-3; logrank test, N = 62). Signature performance analysis was repeated using an optimized threshold (70 th percentile of risks), resulting in HR = 3.38 (p = 1.4E-4; log-rank test, N = 62). The 4-gene signature was validated as predictive of recurrence risk in an independent cohort of patients with resected OSCC and histologically negative margins, and is potentially applicable for clinical decision making on adjuvant treatment and disease monitoring. Oral Squamous Cell Carcinoma (OSCC) is the most common head and neck cancer, and the 8 th most frequent cancer 1 , accounting for >300,000 new cases and >145,000 deaths every year, worldwide 2. Conventional treatment for OSCC includes surgery, radiotherapy and chemotherapy. However, patients often exhibit poor response to treatment and high rates of recurrence, typically between 20-40% 3 , reducing the five-year survival to 50% 4 .

Circulating miR-16-5p, miR-92a-3p, and miR-451a in Plasma from Lung Cancer Patients: Potential Application in Early Detection and a Regulatory Role in Tumorigenesis Pathways

“…Nanostring has been shown to have high sensitivity and specificity and assay reproducibility, which are critical features of clinical utility [40]. More recently, this technology was used to develop a molecular assay that determines the risk of recurrence in oral cancer [41] and triple-negative breast cancer patients [42]. Our results, along with others, suggest that the Nanostring assay is useful for molecular testing development across multiple tumor types, including early detection of lung cancer.…”

“…The Nanostring nCounter ® assay used 200 ng of RNA from each sample. This assay was performed at the Princess Margaret Genomics Centre ( ), according to the manufacturer’s protocol, and as described in previous studies [ 41 ]. The nSolver™ Analysis Software ( ) was used for global normalization of miRNA expression data against positive and negative controls, exogenous and stable endogenous controls, and to calculate fold change (FC) and associated p -values for deregulated miRNAs in patient plasma compared to reference, healthy controls.…”

LncRNA MIR31HG Drives Oncogenicity by Inhibiting the Limb-Bud and Heart Development Gene (LBH) during Oral Carcinoma

“…The roles played by LBH in OSCC pathogenesis remain hitherto obscure. MMP1 and BMP2 are known to modulate cell activity and the tumor microenvironment, as well as being factors associated with OSCC relapse [5,46,47]. This study identifies the up-regulation of MIR31HG in cytobrushed OPMD samples.…”

“…This study has identified a MIR31HG induced differential gene expression profile that is independent of miR-31. MMP1 and BMP2, which are prognostic factors of HN-SCC [5,46,47], and are up-regulated by MIR31HG at the transcription level. During the senescence induced by oncogenic stimulation, cytosolic MIR31HG would seem to increase IL1A protein levels, which then transactivates a panel of secretory molecules such as MMP1 [17].…”

Contribution of Genomics to the Surgical Management and Study of Oral Cancer

“…Microsatellite instability Surgical margins 61 mRNA Nanostring/4-gene signature Surgical margins 64 Methylated DNA Quantitative methylation-specific PCR Surgical margins 65 DNA NGS: missense mutations TP53 (EA score), intratumoral heterogeneity (MATH)…”