Application of a non-invasive oral brushing procedure based on bisulfite sequencing of a 13-gene panel to study high-risk OSCC patients

Pre-Operative Evaluation of DNA Methylation Profile in Oral Squamous Cell Carcinoma Can Predict Tumor Aggressive Potential

“…Previous studies, performed by our group, demonstrated the importance of epigenetic alterations and aberrant DNA methylation of specific genes to discriminate OSCC and its precursors lesions from benign oral mucosal lesions [14][15][16]. Specifically, 19 genes known to be altered in OSCC [14,[17][18][19][20] were evaluated by bisulfite Next Generation Sequencing (NGS) with the aim of developing a noninvasive procedure for OSCC detection based on oral brushing.…”

A 13-Gene DNA Methylation Analysis Using Oral Brushing Specimens as an Indicator of Oral Cancer Risk: A Descriptive Case Report

“…Furthermore, few studies investigate methylation profile in premalignancy with the aim to investigate the predictive value of methylation markers. In particular, some authors demonstrated a significant relationship between the presence of histological dysplasia and an altered methylation profile of multiple genes [ 12 , 33 , 34 , 35 ]. Other authors identified epigenetic markers with predictive power in oral premalignancy.…”

“…We used our recently developed non-invasive procedure consisting of an oral-brushing minimally invasive sampling procedure and DNA methylation analysis of a preselected panel of 13 genes in the oral mucosa. Brushing samples were collected as described previously [ 10 , 12 , 13 , 16 ] at five different times as shown in Table 1 : at the time of OL diagnosis, collected from the surface of the white lesion (December 2016); concomitant with the biopsy, which led to the diagnosis of the index OSCC in the proliferative area with homogeneous dye (April 2019); 6 months after surgical removal of the primitive OSCC in the regenerative area following the primary OSCC resection (October 2019); at the time the secondary tumor appeared, collected from the tumor mass (May 2020); and 6 months after surgical removal of the secondary tumor in the regenerative area following the second OSCC resection (December 2020). Brushing cell collection in the regenerative mucosa after the primary and secondary tumor was performed in the area correspondent the original place of OL: vestibular and lingual gingiva, left cheek, and soft palate.…”

“…A recent multicenter study validated the procedure in an extensive collection of 220 brushing specimens: 93.6% (103/110) of the brushing specimens collected from patients with OSCC lesions were identified in blindness as positive, and 84.9% (90/106) of the samples from healthy individuals were identified as negative [ 11 ]. Subsequently, a 13-gene DNA methylation analysis was applied to the oral brushing samples from high-risk OSCC patients in two subsequent preliminary studies to evaluate the role of our procedure as an indicator of oral cancer risk [ 12 , 13 ]. In the first study, we identified a significant relationship between high-grade dysplasia and positive brushing specimen values collected in patients affected by OL [ 12 ].…”

“…Subsequently, a 13-gene DNA methylation analysis was applied to the oral brushing samples from high-risk OSCC patients in two subsequent preliminary studies to evaluate the role of our procedure as an indicator of oral cancer risk [ 12 , 13 ]. In the first study, we identified a significant relationship between high-grade dysplasia and positive brushing specimen values collected in patients affected by OL [ 12 ]. In the second study, a positive result in an oral brushing specimen collected from regenerating mucosa after resecting OSCC was the most powerful variable related to the appearance of a secondary tumor [ 13 ].…”