Moderate Oral Epithelial Dysplasia Research Articles

P53 Abnormal Oral Epithelial Dysplasias are Associated With High Risks of Progression and Local Recurrence—A Retrospective Study in a Longitudinal Cohort

Grading of oral epithelial dysplasia (OED) can be challenging with considerable intraobserver and interobserver variability. Abnormal immunohistochemical (IHC) staining patterns of the tumor suppressor protein, p53, have been recently shown to be potentially associated with progression in OED. We retrospectively identified 214 oral biopsies from 203 patients recruited in a longitudinal study between 2001 and 2008 with a diagnosis of reactive, nondysplastic lesions, low-grade lesions (mild OED and moderate OED) and high-grade lesions (HGLs; severe OED/carcinoma in situ). Tissue microarrays were constructed from the most representative area of the pathology. Three consecutive sections were sectioned and stained for hematoxylin and eosin, p53 IHC, and p16 IHC. The staining results were reviewed by 2 pathologists blinded to clinical outcome. Samples were categorized into p53 abnormal OED (n = 46), p53 conventional OED (n = 118), and p53 human papillomavirus (HPV) OED (HPV associated) (n = 12) using a previously published pattern-based approach. All cases of p53 HPV OED (HPV associated) were identified in HGLs. In contrast, cases of p53 abnormal OED were observed in mild OED (9.5%), moderate OED (23%), and severe OED/carcinoma in situ (51%). None of the 27 reactive or nondysplastic lesions showed abnormal p53 staining patterns. Among the 135 low-grade lesions, 23 cases (17.0%; 2 mild OEDs and 21 moderate OEDs) progressed to HGL or squamous cell carcinoma, with 11 cases showing progression within the first 3 years. Remarkably, 82% (9/11) of these faster progressors showed abnormal p53 patterns. Survival analysis revealed that p53 abnormal OED had significantly poorer progression-free probability (P < .0001) with hazard ratio of 11.24 (95% CI, 4.26-29.66) compared with p53 conventional OED. Furthermore, p53 abnormal OED had poorer local recurrence–free survival compared with p53 wild-type OED (P = .03). The study supports that OED with p53 abnormal pattern is at high risk for progression and recurrence independent of the dysplasia grade.

Visual Analytics: A Method to Explore Natural Histories of Oral Epithelial Dysplasia.

Risk assessment and follow-up of oral potentially malignant disorders in patients with mild or moderate oral epithelial dysplasia is an ongoing challenge for improved oral cancer prevention. Part of the challenge is a lack of understanding of how observable features of such dysplasia, gathered as data by clinicians during follow-up, relate to underlying biological processes driving progression. Current research is at an exploratory phase where the precise questions to ask are not known. While traditional statistical and the newer machine learning and artificial intelligence methods are effective in well-defined problem spaces with large datasets, these are not the circumstances we face currently. We argue that the field is in need of exploratory methods that can better integrate clinical and scientific knowledge into analysis to iteratively generate viable hypotheses. In this perspective, we propose that visual analytics presents a set of methods well-suited to these needs. We illustrate how visual analytics excels at generating viable research hypotheses by describing our experiences using visual analytics to explore temporal shifts in the clinical presentation of epithelial dysplasia. Visual analytics complements existing methods and fulfills a critical and at-present neglected need in the formative stages of inquiry we are facing.

An Immunohistochemical Study of HIF-1 Alpha in Oral Epithelial Dysplasia and Oral Squamous Cell Carcinoma.

To evaluate and compare the expression of HIF-1 Alpha (HIF-1α) in oral epithelial dysplasia (OED) and various grades of Oral squamous cell carcinoma (OSCC). 30 cases each of OEDand OSCC were stained with HIF-1α antibody. Quantification of HIF-1α positive cellswas carried out and the data was statistically analysed. The mean % HIF-1α labeling index (HIF-1α LI) increased significantly from mild OED (32.11%), moderate OED (55.07%), to severe OED (64.58%). There was a statistically significant increase in the expression of HIF-1α as grades of OED increased. The mean HIF-1α LI % in well differentiated OSCC was 46.3%, Moderately differentiated OSCC-76.31% and Poorly differentiated OSCC-89.9%. The mean HIF-1α LI was found to increase with increasing grades of OSCC which was statistically significant (P < 0.05). Further a comparison of mean HIF-1α LI in OED with different histologic grades of OSCC by Independent samples t test was performed. We found statistically significant difference between OED and moderately differentiated OSCC and OED and poorly differentiated OSCC (P = 0.000). Progressive increase in expression of HIF-1α was noted from OED to OSCC. It can be postulated that epithelial dysplastic lesions with increased HIF-1α expression are at greater risk of malignant transformation, suggesting that the expression of HIF-1α is an early event in oral carcinogenesis.

Similar Effect of P16 Hydroxymethylation and True Methylation on Prediction of Malignant Transformation of Oral Epithelial Dysplasia: A Prospective Study

Background: Total P16 methylation (P16M), including P16 hydroxymethylation (P16H) and true-P16M, correlates with malignant transformation of oral epithelial dysplasia (OED). Both true-P16M and P16H are early events in carcinogenesis. Aim: The aim of this study is to prospectively determine if discrimination of true-P16M from P16H similarly is necessary for prediction of cancer development from OEDs. Methods: Patients (n = 265) with mild or moderate OED were recruited into the double-blind 2-center cohort. Total-P16M and P16H were analyzed using the 115-bp MethyLight, TET-assistant bisulfite (TAB) methylation-specific PCR (MSP), and TAB-sequencing. Total-P16M-positive and P16H-negative samples were defined as true-P16M-positive. Progression of OEDs was monitored for a minimum 24 months follow-up period. Results: P16H was detected in 23 of 73 (31.5%) total-P16M-positive OEDs. Follow-up information was obtained from 247 patients with an ultimate compliance of 93.2%. OED-derived squamous cell carcinomas were observed in 13.0% (32/247) patients during the follow-up (median, 41.0 months). The cancer progression rate for total-P16M-positive patients was significantly increased when compared with total-P16M-negative patients (23.3% vs 8.6%; adjusted odds ratio = 2.67 [95% CI: 1.19-5.99]). However, the cancer progression rate was similar between P16H- and true-P16M-positive OEDs (26.1% [6/23] vs 22.0% [11/50]; odds ratio = 0.80 [95% CI: 0.22-2.92]). The progression-free survival was also similar for these patients. Conclusion: P16H and true-P16M are similar biomarkers for determining malignant potential of OEDs. Discrimination of P16H from true-P16M, at least in OED, may be not necessary in clinical applications.

A similar effect of P16 hydroxymethylation and true-methylation on the prediction of malignant transformation of oral epithelial dysplasia: observation from a prospective study

BackgroundTotal P16 methylation (P16M), including P16 hydroxymethylation (P16H) and true-P16M, correlates with malignant transformation of oral epithelial dysplasia (OED). Both true-P16M and P16H are early events in carcinogenesis. The aim of this study is to prospectively determine if discrimination of true-P16M from P16H is necessary for prediction of cancer development from OEDs.MethodsPatients (n = 265) with mild or moderate OED were recruited into the double blind two-center cohort. Total-P16M and P16H were analyzed using the 115-bp MethyLight, TET-assisted bisulfite (TAB) methylation-specific PCR (MSP), and TAB-sequencing. Total-P16M-positive and P16H-negative samples were defined as true-P16M-positive. Progression of OEDs was monitored for a minimum 24 months follow-up period.ResultsP16H was detected in 23 of 73 (31.5%) total-P16M-positive OEDs. Follow-up information was obtained from 247 patients with an ultimate compliance rate of 93.2%. OED-derived squamous cell carcinomas were observed in 13.0% (32/247) patients during follow-up (median, 41.0 months). The cancer progression rate for total-P16M-positive patients was significantly increased when compared to total-P16M-negative patients [23.3% vs 8.6%; adjusted odds ratio = 2.67 (95% CI: 1.19–5.99)]. However, the cancer progression rates were similar between P16H- and true-P16M-positive OEDs [26.1% (6/23) vs 22.0% (11/50); odds ratio = 0.80 (95% CI: 0.22–2.92)]. The cancer-free survival was also similar for these patients.ConclusionP16H and true-P16M are similar biomarkers for determining malignant potential of OEDs. Discrimination of P16H from true-P16M, at least in OED, may be not necessary in clinical applications.Trial registrationThis study is registered prospectively in the U.S. National Institutes of Health Clinical Trials Protocol Registration System (trial number NCT02967120, available at https://ClinicalTrials.gov/ct2/show/NCT02967120).

Coalition of E-cadherin and vascular endothelial growth factor expression in predicting malignant transformation in common oral potentially malignant disorders.

Background:Reduced E-cadherin expression and increased VEGF expression is known to be involved in tissue growth and transformation of Oral Potentially Malignant Disorders (OPMDs) and has been correlated with their differing histological grades in numerous studies.Aim:To evaluate Immunohistochemical (IHC) expression of both E-cadherin and VEGF in predicting the malignant transformation potential of common OPMDs.Materials and Methods:Ten cases each of Normal Oral mucosa (NOM), Mild Oral Epithelial Dysplasia (OED), Moderate OED, Severe OED, Oral Submucous Fibrosis, (OSMF) and Oral Squamous Cell Carcinoma (OSCC) were stained and evaluated for the expression of Ecadherin and VEGF. Quick score (QS) for expression intensity in all epithelial layers was calculated for both markers and results statistically analysed using Kruskal –Wallis ANOVA and Mann-Whitney “U” test.Results:E-cadherin expression was continuous and membranous in all the layers of NOM and reduced with progressing grades of OED to OSCC. In OSMF, expression was intermediate between moderate and severe OED. VEGF expression increased as the disease progressed from normal to increasing grades of OED to malignancy. In OSMF, expression was similar to that in mild OED. VEGF, E-cadherin expression for basal and parabasilar cells showed a strong statistically significant negative correlation in NOM. A very strong statistically significant positive correlation with perfect monotonic relation was noted in superficial cells in severe OED group and OSCC group.Conclusion:E-Cadherin and VEGF could be used as combination markers to predict the potential risk for malignant transformation in OEDs.

Abstract 3265: Characterization of epithelial oral dysplasia in non-smokers: working towards precision medicine

Abstract Objectives: Although tobacco usage is still one of the strongest risk factors associated with oral cancer risk, there is a subset of non-smokers who develop oral cancer. Tobacco cessation efforts have resulted in a drop in oral cancer rates associated with this habit, leading to a growing interest in the increased proportion of cases occurring among non-smokers (NS). Lesions with oral epithelial dysplasia (OED) are at risk of progressing to oral cancer. Not only is the natural history of OED in NS poorly understood, but the path to interception of disease in NS is poorly defined. There is a gap in the knowledge surrounding the clinicopathological and genetic characterization, and the risk of progression in this growing category. This information is critical to the evolution of precision medicine in this subgroup. The aim of this study was to: 1) Describe the molecular and clinicopathological features of OED in NS as compared to smokers in longitudinal follow-up; and 2) To compare progression rates and time to progression in NS and smokers with OED. Methods: The study focused on cases with histologically confirmed mild or moderate OED in follow-up in the Oral Cancer Prediction Longitudinal Study. Clinicopathological data, including lesion site, size, texture, colour, consistency, border characteristics, fluorescence visualization (FV), and toluidine blue (TB) retention, were collected in addition to detailed smoking history. A genomic based marker test (gMART) which uses loss of heterozygosity (LOH) at key chromosomal loci to stratify lesions to progression risk, was performed on baseline biopsies. Progression was considered to be advancement to severe dysplasia, carcinoma in situ, or squamous cell carcinoma. Results: Out of 231 OED, 30% were NS related, based on self-reported smoking status. Although there were more smokers with OED than NS, a significantly higher proportion of the OED underwent malignant transformation in NS (P=.048). Although not significant, time to outcome was also faster in this group. Most clinical features were equally predictive except for lesion site. Ever smokers (ES) were more likely to have OED at the floor of mouth while OED was more likely to occur at the tongue or gingiva (P&amp;lt;.001) in NS. LOH risk patterns were strongly associated with progression (moderate risk = OR 4.84; high risk = OR 28.1; P=.001) and equally sensitive in both NS and ES subgroups. Conclusions: These findings support the premise that progressive lesions in NS and ES have some similar genetic underpinnings, and emphasize the need for clinicians to consider the molecular genomic profiles in the triage of OED. LOH markers can sort high-risk lesions, despite risk habits, and should be an important consideration in the treatment of OED. This marker is of particular use in the targeting of candidates for chemoprevention. Citation Format: Leigha D. Rock, Miriam P. Rosin, Lewei Zhang, Batoul Shariati, Denise M. Laronde. Characterization of epithelial oral dysplasia in non-smokers: working towards precision medicine [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 3265. doi:10.1158/1538-7445.AM2017-3265

Langerhans cell counts in oral epithelial dysplasia and their correlation to clinicopathological parameters.

Langerhans cells (LCs) are antigen presenting cells. This study assessed the LC counts in oral epithelial dysplasia (OED) and their correlation to clinicopathological parameters. This study examined the LC counts in the epithelia and subepithelial connective tissues of 58 patients with OED (21 mild, 18 moderate, and 19 severe OED lesions) and 10 specimens of normal oral mucosa (NOM) by anti-S-100 protein immunostaining. We found that the mean LC counts in the epithelia or subepithelial connective tissues increased significantly from NOM samples through mild and moderate OED to severe OED samples. In addition, a significant correlation was found between higher mean LC counts in the dysplastic epithelia of OED samples and OED lesions with thicker epithelial layers (p<0.001) or wider inflammatory zones (p<0.001), and between higher mean LC counts in the subepithelial connective tissues of OED samples and OED lesions with wider inflammatory zones (p<0.001). Moreover, the nine OED lesions with malignant transformation had a significantly lower mean LC count than the 49 OED lesions without malignant transformation. The significant and gradual elevation in LC count from NOM through mild and moderate OED to severe OED lesions suggests an upregulation of immunosurveillance ability in OED patients during the early oral carcinogenesis process. A low LC count in OED lesions may suggest the partial loss of immunosurveillance ability against dysplastic cells; this in turn favors the malignant transformation of an OED lesion into oral cancer.

P16 Methylation as an Early Predictor for Cancer Development From Oral Epithelial Dysplasia: A Double-blind Multicentre Prospective Study

BackgroundSilencing of P16 through methylation and locus deletion is the most frequent early events in carcinogenesis. The aim of this study is to prospectively determine if early P16 methylation is a predictor for oral cancer development. MethodsPatients (n=181) with mild or moderate oral epithelial dysplasia (OED) were recruited into the double blind multicentre cohort. P16 methylation was analyzed using the MethyLight assay. Progression of OEDs was monitored for a minimum 3year follow-up period. FindingsP16 methylation-informative cases (n=152) were enrolled in the prospective multicenter cohorts with an ultimate compliance of 96.7%. OED-derived squamous cell carcinomas were observed in 21 patients (14.3%) during the follow-up (median, 41.0months). The cancer progression rate from the P16 methylation-positive patients was significantly increased when compared to P16 methylation-negative patients [27.1% vs 8.1%; adjusted odds ratio=4.6; P=0.006]. When the P16 methylation-positive criteria were used as a biomarker for early prediction of cancer development from OEDs, sensitivity and specificity of 62% and 76% were obtained, respectively. InterpretationP16 methylation is unequivocally a marker for determining the malignant potential of OED and there is no need for further research regarding this aspect. FundingNational Basic Research Programs of China (2011CB504201 and 2015CB553902), Beijing Science and Technology Commission (Z090507017709016), and Beijing Municipal Administration of Hospital (XM201303) to Dajun Deng. The funding agencies have no role in the actual experimental design, patient recruitment, data collection, analysis, interpretation, or writing of this manuscript.

Methylation of p16 CpG Island Associated with Malignant Progression of Oral Epithelial Dysplasia: A Prospective Cohort Study

Inactivation of p16 gene by CpG methylation is a frequent event in oral epithelial dysplasia. To investigate the predictive value of p16 methylation on malignant potential in oral epithelial dysplasia, we carried out the prospective cohort study. One hundred one patients with histologically confirmed mild or moderate oral epithelial dysplasia were included in the present cohort study. p16 Methylation status of the oral epithelial dysplasia lesions from 93 cases was obtained by methylation-specific PCR. Progression of the oral epithelial dysplasia lesions was examined in 78 cases histologically during a 45.8 months follow-up period. The association between p16 methylation and progression of oral epithelial dysplasia was analyzed. Of the 93 enrolled cases, 15 cases were lost during the follow-up because of changes of contact information, with a compliance of 83.9%. p16 Methylation was detectable in oral epithelial dysplasia lesions from 32 (41.0%) of 78 enrolled patients. Oral epithelial dysplasia-related squamous cell carcinomas were observed in 22 patients (28.2%) during the follow-up. Rate of progression to oral cancer in patients with the p16-methylated oral epithelial dysplasia was significantly higher than that with the p16-unmethylated oral epithelial dysplasia (43.8% versus 17.4%; adjusted odds ratio, 3.7; P = 0.013), especially for patients at the baseline age of > or = 60 years (adjusted odds ratio, 12.0; P = 0.003) and patients with moderate oral epithelial dysplasia (adjusted odds ratio, 15.6; P = 0.022). The overall sensitivity and specificity of prediction of malignant transformation of oral epithelial dysplasia by p16 methylation were 63.6% and 67.9%, respectively. p16 Methylation was correlated with malignant transformation of oral epithelial dysplasia and is a potential biomarker for prediction of prognosis of mild or moderate oral epithelial dysplasia.

Outcome of oral dysplasia: a retrospective hospital‐based study of 207 patients with a long follow‐up

The aim of this retrospective hospital-based study was to review and evaluate the long-term outcome of patients with oral epithelial dysplasia (OED), with or without surgical intervention, to identify factors affecting clinical course and malignant evolution. Patients with a follow-up of at least 12 months were included. Data collected were statistically analyzed. The mean age was 63.58 years for women (n = 100) and 64.17 years for men (n = 107). One hundred and thirty-five of the patients had lesions with histopathological features of mild OED, 50 had moderate OED and 22 had severe OED. Gender and risk factors seemed not to be related with the development of OED. One hundred and thirty-three patients underwent active treatment. During the period considered, 39.4% of the 207 lesions disappeared; 19.66% remained stable and 33.7% of the total cases showed a new dysplastic event after treatment. Fifteen (7.24%) out of 207 developed a squamous cell carcinoma during follow-up. Our data showed that speckled lesions are more often associated with high histological grade. The risk of malignant development does not seem to be predictable. Surrounded by the limitations of the retrospective designs, we have showed that there is no eminent benefit of surgical intervention of OED in preventing recurrences and malignant development.

Angiogenic Squamous Dysplasia in Oral Epithelial Dysplastic Lesions

Statement of problem: Angiogenic squamous dysplasia (ASD), a qualitative distinct form of angiogenesis, was first described in pre-invasive bronchial mucosa of high-risk individuals. It is characterized histologically by the presence of capillary tufts that are closely juxtaposed to and projecting into the dysplastic bronchial epithelium. Objective: To determine whether ASD occurs in oral epithelial dysplastic lesions. Methods: Sixty cases of potentially malignant oral epithelial lesions comprising 20 mild epithelial dysplasia (ED), 20 moderate ED and 20 severe ED(inclusive of carcinoma-in-situ), and 10 normal oral mucosa (as normal controls) were retrieved from the archives of the Department of Oral Pathology, Oral Medicine & Periodontology, Faculty of Dentistry, University of Malaya, and the Cancer Research Centre, Institute for Medical Research, Kuala Lumpur, Malaysia. The grading of oral ED was in accordance with the recommendations of the World Health Organization. For all selected cases, new 5 micron thick sections were prepared for H&E staining, and for immunohistochemistry with three vascular markers, CD31, CD34 and CD105, with appropriate positive and negative controls. The established criteria for identification of ASD was applied. Results: ASD was identified in parts of severe oral ED, but was absent in normal oral epithelium, mild and moderate oral ED. About 65% of these severe oral ED cases were from individuals with high-risk habits. As with bronchial ASD, the capillaries typically formed CD31- and CD34-positive projections or loops that abut onto the overlying dysplastic oral epithelium causing the latter to assume a papillary surface configuration. Unlike bronchial ASD which occurs in respiratory-type epithelium, oral ASD was found in keratinzed stratified squamous epithelium. CD105 confirmed the presence of neoangiogenesis. Conclusions: Present findings confirm that ASD can occur in oral severe ED. It also demonstrates that this angiogenic abnormality is not unique to bronchial mucosal dysplastic lesions.

Evaluation of glutathione S-transferase activity in human buccal epithelial dysplasias and squamous cell carcinomas

Glutathione S-transferase (GST) activity and amount of GST α, μ and π isoforms were measured in 40 patients with histopathologically confirmed oral epithelial dysplasia (OED) and squamous cell carcinoma of buccal mucosa. The results were compared with those of normal mucosa in an equal number of age- and sex-matched healthy controls. Mean total GST activities were significantly elevated from normal buccal mucosa for mild OED, moderate OED, severe OED and squamous cell carcinoma. GST activity of value approximating 100 nmol/min/mg distinguished between normal and dysplasia, and of value about 400 nmol/min/mg delineated between dysplasia and squamous cell carcinoma were observed. GST π was the predominant class in both the diseased and normal buccal mucosa examined. This class π GST was present at an intracellular concentration, which was significantly higher in diseased buccal mucosa than in normal buccal mucosa. These results indicated that π class GST was the major form of this enzyme in the cytosolic fraction of oral mucosa. The severity of OED related to squamous cell carcinoma development seemed to increase concomitantly with an increase in the level of this enzyme. Further studies will validate the role of GST π estimation in predicting the potential malignancy of OED.