Abstract
Simple SummaryIncisional tissue biopsy is highly invasive, but it remains the current practice for oral cancer diagnosis through histopathological interpretation. To reduce biopsy invasiveness without compromising diagnosis, we pioneered a multigene RT-qPCR method for cancer detection using only a tiny 1 mm3 minimally invasive biopsy. Here we presented international multicohort validation of our second-generation method which involved new genes from matrix/stroma and immune regulations enabling sensitive, quantitative and precise oral cancer detection in otherwise ambiguous oral lesions.Background: Heterogeneity in oral potentially malignant disorder (OPMD) poses a problem for accurate prognosis that impacts on treatment strategy and patient outcome. A holistic assessment based on gene expression signatures from both the tumour cells and their microenvironment is necessary to provide a more precise prognostic assessment than just tumour cell signatures alone. Methods: We reformulated our previously established multigene qPCR test, quantitative Malignancy Index Diagnostic System (qMIDS) with new genes involved in matrix/stroma and immune modulation of the tumour microenvironment. An algorithm calculates and converts a panel of 16 gene mRNA expression levels into a qMIDS index to quantify risk of malignancy for each sample. Results: The new qMIDSV2 assay was validated in a UK oral squamous cell carcinoma (OSCC) cohort (n = 282) of margin and tumour core samples demonstrating significantly better diagnostic performance (AUC = 0.945) compared to previous qMIDSV1 (AUC = 0.759). Performance of qMIDSV2 were independently validated in Chinese (n = 35; AUC = 0.928) and Indian (n = 95; AUC = 0.932) OSCC cohorts. Further, 5-year retrospective analysis on an Indian dysplastic lesion cohort (n = 30) showed that qMIDSV2 was able to significantly differentiate between lesions without transformation and those with malignant transformation. Conclusions: This study validated a novel multi-gene qPCR test on a total of 535 tissue specimens from UK, China and India, demonstrating a rapid minimally invasive method that has a potential application for dysplasia risk stratification. Further study is required to establish if qMIDSV2 could be used to improve OPMD patient management, guide treatment strategy and reduce oral cancer burden.
Highlights
This study investigated the possibility to evolve and improve our original qMIDSV1 assay [24,30] by including new genes with functions in stroma/matrix and immune regulation to enhance diagnostic performance for early oral cancer detection and/or to predict malignant transformation in oral potentially malignant disorder (OPMD)
Since our first publication validating the use of quantitative Malignancy Index Diagnostic System (qMIDS) for early oral squamous cell carcinoma (OSCC) diagnosis [24], we have accumulated large number of qMIDS data on 1761 tissue fragment samples from normal and disease
We subjected our dataset to three methods of sub-groupings prior to linear and polynomial curve-fitting methods to access how well each gene correlated with qMIDSV1 index values (Supplementary Materials Figure S3A)
Summary
The global cancer statistics 2020 (GLOBOCAN 2020) indicated oral cancer (including lip, C00–C06) as one of the eight leading cancer types for both incidence and mortality in men [1]. Majority of OSCC patients have some form of pre-existing oral potentially malignant disorder (OPMD) lesions amenable to early diagnosis and risk stratification [4–8]. Most if not all OPMD patients are either put on regular surveillance or are biopsied and put on a variable period of review before discharge [4–6,8,12]. For the low-risk OPMD patients, regular review will result in unnecessary health service costs and may cause continued anxiety for the patients. For the high-risk patients (
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