DNA aneuploidy with image cytometry for detecting dysplasia and carcinoma in oral potentially malignant disorders: A prospective diagnostic study

Abstract

Background Current evidence on diagnostic value of aneuploidy with DNA image cytometry (ICM) using brushings for oral potentially malignant disorders (OPMD) is limited by sample size and inconsistent classification criteria of aneuploidy. This study aimed to explore the optimal cut‐off values of DNA content and evaluate the diagnostic accuracy of DNA‐ICM for detecting dysplasia and/or carcinoma in OPMD. Materials and Methods A total of 401 consecutive patients with OPMD were enrolled in this prospective diagnostic study. Brushing and biopsy sample form each patient was processed by DNA‐ICM and histological examination respectively. Results When the optimal cut‐off of at least one aneuploid cell with DNA index (DI) ≥2.3, the area under the curves (AUC) was 0.735 and positive predictive value was 92.7% for detecting dysplasia within OPMD. When the optimal cut‐off of at least one aneuploid cell with DI ≥ 3.5, the AUC was 0.851 and negative predictive values was 96.8% for detecting carcinoma within OPMD. Importantly, multivariate analysis revealed that aneuploidy with DI ≥ 2.3 in OPMD was significantly associated with dysplasia risk (adjusted OR, 5.52; 95%CI, 2.90‐10.51; P < .001), and aneuploidy with DI ≥ 3.5 in OPMD was strongly associated with malignant risk (adjusted OR, 21.05; 95%CI, 9.34‐47.41; P < .001). Conclusions This largest‐scale diagnostic study optimized the criteria of aneuploidy cytology for noninvasive detection of oral dysplasia and carcinoma within OPMD. DNA aneuploidy in OPMD was an independent marker that strongly associated with oral dysplasia/carcinoma. Our findings suggest that DNA‐ICM may serve as a useful noninvasive adjunctive tool for oral cancer and OPMD screening.

Aneuploidy and loss of heterozygosity as risk markers for malignant transformation in oral mucosa

“…Several groups have confirmed the value of DNA ploidy as a predictor of malignant transformation (Bradley et al, 2010;Bremmer et al, 2011;Siebers et al, 2013;Sperandio et al, 2013;Torres-Rendon et al, 2009;Zaini et al, 2018). Abnormal DNA ploidy correlates with the presence and severity of dysplasia (Donadini et al, 2010;Grässel-Pietrusky et al, 1982;Pentenero et al, 2009;Saito et al, ,1995Saito et al, , , 1998van Zyl et al, 2012), is more frequently found in higher grades of dysplasia (Li et al, 2020;Sperandio et al, 2013;van Zyl et al, 2012) and in lesions considered high-risk clinically (Xiao et al, 2015;van Zyl et al, 2012). Correlation with dysplasia is not necessarily an advantage since the predictive value of dysplasia is reported to be extremely variable, but DNA ploidy is able to detect changes in OPMD that have minimal or no dysplasia (Sperandio et al, 2013;Zaini et al, 2018), as can flow cytometry (Pentenero et al, 2012).…”

“…Very small amounts of tissue are required for analysis. Both flow cytometry and image cytometry can be applied to scrapings of mucosa, endoscopic forceps biopsy, brush biopsy or other cytological specimens (Cowpe et al, 1985;Dineshkumar et al, 2019;Kammerer et al, 2013;Pektas et al, 2006;Remmerbach et al, 2001Remmerbach et al, , 2003Velleuer et al, 2020) and results match those on incisional biopsy (Li et al, 2020;Maraki et al, 2004) and correlate with clinical risk F I G U R E 1 Typical DNA ploidy histograms obtained by image-based cytometry. Y-axis total nuclei analysed and X-axis optical density in Feulgen stain is directly proportional to DNA content, calibrated as c value (2c = diploid).…”

“…The second major factor in predictive value is diagnostic threshold. Accepting a very low threshold for aneuploidy produces a very sensitive positive predictive value but very low negative predictive value (Li et al, 2020). An third factor that may alter predictive values is that some publications use a system that combines DNA aneuploidy with an element of cytology to make a final diagnosis (Bradley et al, 2010).…”

“…Whether patterns of ploidy such as high 5c exceeding rates, multiple stem lines or diploid index could be used to improve diagnostic criteria is not clear. So far, few studies have addressed these issues (Bradley et al, 2010;Giaretti et al, 2013;Seoane et al, 1998), but suggest that peaks of higher diploid index carry higher risk and correlate better with dysplasia (Li et al, 2020). Combining the DNA ploidy result with dysplasia grade (Zaini et al, 2018), other biomarkers (Fleskens et al, 2010;Ogden et al, 1994) and clinical information (Castagnola et al, 2017) appears to provide additional predictive information but has less value when transformation rates in the population are very high (Sathasivam et al, 2020).…”

A Systematic Review of Oral Biopsies, Sample Types, and Detection Techniques Applied in Relation to Oral Cancer Detection

“…The following supporting information can be downloaded at: , Table S1: Summary characteristics of included studies [ 190 , 191 , 192 , 193 , 194 , 195 , 196 , 197 , 198 , 199 , 200 , 201 , 202 , 203 , 204 , 205 , 206 , 207 , 208 , 209 , 210 , 211 , 212 , 213 , 214 , 215 , 216 , 217 , 218 , 219 , 220 , 221 , 222 , 223 , 224 , 225 , 226 , 227 , 228 , 229 , 230 , 231 , 232 , 233 , 234 , 235 , 236 , 237 , 238 , 239 , 240 , 241 , 242 <...…”