Oral Potentially Malignant Lesions Research Articles

Micronuclei detection in oral cytologic smear: does it add diagnostic value?

BackgroundScreening and early diagnosis of oral squamous cell carcinoma (OSCC) are directly associated with increased survival rate and improved prognosis. Noninvasive diagnostic tools have been implemented in the early detection as toluidine blue staining, optical imaging, and oral cytology. This study aimed to assess and compare the presence of micronuclei (MN) in oral exfoliative cytology of healthy controls, subjects exposed to high-risk factors for oral cancer, subjects with oral potentially malignant lesions (OPMLs), and those with malignant oral lesions.Subjects and methodsA total number of 92 subjects were divided into 46 healthy controls with no oral mucosal lesions (23 with no evidence of cancer risk factors and 23 with cancer risk factors), 23 with OPMLs and 23 with oral malignant lesions. All the 92 participants were subjected to cytological sampling for detection of MN. The final diagnosis of the oral lesions was confirmed by the histopathological picture and compared to the cytological results.ResultsThe results showed that the diagnostic accuracy of MN was higher in OPMLs group (95.2%). The sensitivity of MN test in malignant group was much lower (52.2%); however, all the cytological criteria of malignancy were markedly detected as compared to the OPMLs group.ConclusionsConventional oral cytology supported by MN is highly beneficial as adjunctive tool in the screening for early detection of dysplastic oral lesions.

Accuracy of Remote Examination for Detecting Potentially Malignant Oral Lesions: A Systematic Review and Meta-Analysis.

Objective: We aimed to investigate the accuracy of remote examination by photographs compared to in-person clinical examination for detecting potentially malignant oral lesions (PMOLs). Methods: The Reporting Guide and Guidelines for Writing Systematic Reviews (Preferred Reporting Items for Systematic Reviews and Meta-Analysis [PRISMA]) guided the reporting of findings. The search was conducted by two independent reviewers in six databases with no language restriction until November 2022. The Population, Test-Index, Reference Standard, Outcome and Study Design (PIROS) strategy guided the eligibility criteria, and studies with adult patients (P) examined remotely (I) and in-person (R) to verify the detection of PMOLs (O) were considered. The methodological quality was assessed by QUADAS-2, and the certainty of the evidence was measured by the Grading of Recommendations, Assessment, Development, and Evaluation (GRADE). Results: A total of 769 files were identified. After removing duplicates and reading titles and abstracts, 16 were read in full, from which 6 articles then comprised the qualitative synthesis. The oral clinical examination was the reference standard in four studies. Five studies presented high risk of bias in at least one assessment domain. A high probability of detection of PMOL by remote examination (97.37%) was observed for the three studies included in the meta-analysis, which presented high heterogeneity among them. The certainty of evidence for the outcome was considered very low. Conclusions: Remote tools for detecting PMOLs may be feasible and assertive, but new studies are required to incorporate them into clinical practice. Clinical Relevance: Remote examination for the detection of PMOLs has the potential to favoring the early diagnosis of malignant lesions.

Early Diagnosis of Oral Cancer and Lesions in Fanconi Anemia Patients: A Prospective and Longitudinal Study Using Saliva and Plasma.

Fanconi anemia (FA) patients display an exacerbated risk of oral squamous cell carcinoma (OSCC) and oral potentially malignant lesions (OPMLs) at early ages. As patients have defects in their DNA repair mechanisms, standard-of-care treatments for OSCC such as radiotherapy and chemotherapy, give rise to severe toxicities. New methods for early diagnosis are urgently needed to allow for treatment in early disease stages and achieve better clinical outcomes. We conducted a prospective, longitudinal study wherein liquid biopsies from sixteen patients with no clinical diagnoses of OPML and/or OSCC were analyzed for the presence of mutations in cancer genes. The DNA from saliva and plasma were sequentially collected and deep-sequenced, and the clinical evaluation followed over a median time of approximately 2 years. In 9/16 FA patients, we detected mutations in cancer genes (mainly TP53) with minor allele frequencies (MAF) of down to 0.07%. Importantly, all patients that had mutations and clinical follow-up data after mutation detection (n = 6) developed oral precursor lesions or OSCC. The lead-time between mutation detection and tumor diagnosis ranged from 23 to 630 days. Strikingly, FA patients without mutations displayed a significantly lower risk of developing precursor lesions or OSCCs. Therefore, our diagnostic approach could help to stratify FA patients into risk groups, which would allow for closer surveillance for OSCCs or precursor lesions.

Use of DNA image cytometry in conducting oral cancer screening in rural India

Oral cancer screening can assist in the early detection of oral potentially malignant lesions (OPMLs) and prevention of oral cancers. It can be challenging for clinicians to differentiate OPMLs from benign conditions. Adjunct screening tools such as fluorescence visualisation (FV) and DNA image cytometry (DNA-ICM) have shown success in identifying OPMLs in high-risk clinics. For the first time we aimed to assess these technologies in Indian rural settings and evaluate if these tools helped clinicians identify high-risk lesions during screening. Dental students and residents screened participants in five screening camps held in villages outside of Hyderabad, India, using extraoral, intraoral, and FV examinations. Lesion and normal tissue brushings were collected for DNA-ICM analysis and cytology. Of the 1116 participants screened, 184 lesions were observed in 152 participants. Based on white light examination (WLE), 45 lesions were recommended for biopsy. Thirty-five were completed on site; 25 (71%) were diagnosed with low-grade dysplasias (17 mild, 8 moderate) and the remaining 10 showed no signs of dysplasia. FV loss was noted in all but one dysplastic lesion and showed a sensitivity of 96% and specificity of 17%. Cytology combined with DNA-ICM had a sensitivity of 64% and specificity of 86% in detecting dysplasia. DNA-ICM combined with cytology identified the majority of dysplastic lesions and identified additional lesions, which were not considered high-risk during WLE and biopsy on site. Efforts to follow-up with these participants are ongoing. FV identified most high-risk lesions but added limited value over WLE.

Cell Block Method: An Imperative Tool for Cytological Diagnosis of Oral Potentially Malignant Lesions

Globally, oral squamous cell carcinoma (OSCC) is one of the commonest reported malignancies usually arising from oral potentially malignant lesions (OPMLs) such as leukoplakia, erythroplakia, oral submucous fibrosis (OSMF) and oral lichen planus (OLP). Hence timely and early diagnosis of these disorders is of prime importance to halt their malignant transformation. A search of published works was done using the online databases of PubMed, Scopus, Web of Science, and Cochrane Library”. Data from resource-constrained laboratory settings worldwide shows limited documentation of the efficacy of advanced cytological techniques, including LBC and cell block preparations. The oral mucosa can be a suitable area for regular cytological screening due to its easy accessibility. Liquid-based cytology (LBC) can preserve the cellular details and reduce the overlapping of cells, enabling precise interpretation, reducing false-negative results and aiding in the diagnosis of premalignant and malignant lesions of the oral cavity with more accuracy compared to exfoliative cytology. The remaining sample in LBC can be used in cell block formation and various ancillary tests like immunocytochemistry, immunofluorescence, and molecular studies. Literature showed a scarcity of data available regarding Pakistan. Therefore, the review is aimed to explore the cell block method as a minimally invasive technique for reducing morbidity and mortality associated with OSCC. Keywords: Cell Biology; Oral Squamous Cell Carcinoma; Malignancy.

Predicting Progression of Low-Grade Oral Dysplasia Using Brushing-Based DNA Ploidy and Chromatin Organization Analysis.

Most oral cancers arise from oral potentially malignant lesions, which show varying grades of dysplasia. Risk of progression increases with increasing grade of dysplasia; however, risk prediction among oral low-grade dysplasia (LGD), that is, mild and moderate dysplasia can be challenging as only 5%-15% transform. Moreover, grading of dysplasia is subjective and varies with the area of the lesion being biopsied. To date, no biomarkers or tools are used clinically to triage oral LGDs. This study uses a combination of DNA ploidy and chromatin organization (CO) scores from cells obtained from lesion brushings to identify oral LGDs at high-risk of progression. A total of 130 lesion brushings from patients with oral LGDs were selected of which 16 (12.3%) lesions progressed to severe dysplasia or cancer. DNA ploidy and CO scores were analyzed from nuclear features measured by our in-house DNA image cytometry (DNA-ICM) system and used to classify brushings into low-risk and high-risk. A total of 57 samples were classified as high-risk of which 13 were progressors. High-risk DNA brushing was significant for progression (P = 0.001) and grade of dysplasia (P = 0.004). Multivariate analysis showed high-risk DNA brushing showed 5.1- to 8-fold increased risk of progression, a stronger predictor than dysplasia grading and lesion clinical features. DNA-ICM can serve as a non-invasive, high-throughput tool to identify high-risk lesions several years before transformation. This will help clinicians focus on such lesions whereas low-risk lesions may be spared from unnecessary biopsies.Prevention Relevance: DNA ploidy and chromatin organization of cells collected from oral potentially malignant lesions (OPMLs) can identify lesions at high-risk of progression several years prior. This non-invasive test would enable clinicians to triage high-risk (OPMLs) for closer follow-up while low-risk lesions can undergo less frequent biopsies reducing burden on healthcare resources.

Prevalence and determinants of oral potentially malignant lesions using mobile health in a rural block, northeast India.

In a rural block in North East India, community health workers (CHW) empowered with a mobile phone-based application screened a total of 2,686 participants for Oral Potentially Malignant Lesions (OPMLs), and an oral medicine specialist recommended treatment remotely. Independent risk factors were determined using independent multiple logistic regression models. Nearly 700 (26%) participants were identified with OPMLs. The sensitivity, specificity, positive predictive values, negative predictive values and accuracy of the CHW was 70.3, 88.4, 66.8, 89.9% and 83.7% respectively. Male gender, married status, smokeless tobacco, paan, areca-nut and alcohol consumption were independent predictors of OPMLs, the burden of which in North East India can be attributed to the high consumption of tobacco and non-tobacco products. Such programmes, with the recommendations from remote specialists, will facilitate early detection in remote settings.

Bimodal multispectral imaging system with cloud-based machine learning algorithm for real-time screening and detection of oral potentially malignant lesions and biopsy guidance.

.Significance: Screening and early detection of oral potentially malignant lesions (OPMLs) are of great significance in reducing the mortality rates associated with head and neck malignancies. Intra-oral multispectral optical imaging of tissues in conjunction with cloud-based machine learning (CBML) can be used to detect oral precancers at the point-of-care (POC) and guide the clinician to the most malignant site for biopsy.Aim: Develop a bimodal multispectral imaging system (BMIS) combining tissue autofluorescence and diffuse reflectance (DR) for mapping changes in oxygenated hemoglobin () absorption in the oral mucosa, quantifying tissue abnormalities, and guiding biopsies.Approach: The hand-held widefield BMIS consisting of LEDs emitting at 405, 545, 575, and 610 nm, 5MPx monochrome camera, and proprietary Windows-based software was developed for image capture, processing, and analytics. The DR image ratio (R610/R545) was compared with pathologic classification to develop a CBML algorithm for real-time assessment of tissue status at the POC.Results: Sensitivity of 97.5% and specificity of 92.5% were achieved for discrimination of OPML from patient normal in 40 sites, whereas 82% sensitivity and 96.6% specificity were obtained for discrimination of abnormal (OPML + SCC) in 89 sites. Site-specific algorithms derived for buccal mucosa (27 sites) showed improved sensitivity and specificity of 96.3% for discrimination of OPML from normal.Conclusions: Assessment of oral cancer risk is possible by mapping of absorption in tissues, and the BMIS system developed appears to be suitable for biopsy guidance and early detection of oral cancers.

RARβ Expression in Keratinocytes from Potentially Malignant Oral Lesions: The Functional Consequences of Re-Expression by De-Methylating Agents.

Simple SummaryPatients may develop white or red patches of the lining of the mouth with an increased risk of developing oral cancer. Treatment with Vitamin A derivatives (retinoids) results in some improvement in these lesions, but this is not maintained, and there are side effects. We know that the cells of the mouth lose cellular receptors for retinoids as these lesions develop, initially by a reversible alteration to the DNA (DNA methylation). Drugs, such as 5-AZA-CdR, which reduce DNA methylation, may restore sensitivity to the effects of retinoids. Treatment of a panel of cells from mouth precancer white patches with retinoids, 5-AZA-CdR and a combination results in varied responses: some cells re-sensitise to retinoids, whereas in others, the main effects on cell division rate and cell lifespan seem related to the effects of 5-AZA-CdR alone. These findings help us to understand the varied responses to retinoids in the clinical setting.Loss of RARβ2 expression by promoter methylation is an early event in oral carcinogenesis. Understanding the mechanisms and consequences of RARβ loss may aid in understanding the disappointing results of retinoid chemoprevention trials. This study aimed to describe the effects of all-trans retinoic acid (ATRA) and the de-methylating agent 5-Aza-2′ deoxycytidine (5-AZA-CdR) on a panel of immortal potentially malignant oral lesion (PMOL) cell cultures. RARβ expression was assessed in PMOL tissues by immunohistochemistry. Cells were treated with ATRA ± 5-AZA-CdR, and the effects on the cell cycle and senescence were assessed. In PMOL tissues, RARβ expression was variable, but lower in biopsies which gave rise to immortal cell cultures. Treatment of iPMOL cells with ATRA resulted in little change in RARβ expression, but the addition of 5-AZA-CdR resulted in significant increases. The effects on the cell cycle and senescence were variable and may be related to 5-AZA-CdR, as this has wider effects on the cell cycle. Overall, the response of iPMOL cells to ATRA and 5-AZA-CdR treatment was variable and is dependent on several factors, including RARβ-promoter methylation. These findings may help to explain the lack of consistent effect of retinoids in PMOLs seen in chemoprevention trials.

Validation of a Point-of-Care Optical Coherence Tomography Device with Machine Learning Algorithm for Detection of Oral Potentially Malignant and Malignant Lesions.

Simple SummaryEarly detection is crucial towards improving survival in patients diagnosed with oral cancer. Non-invasive strategies equivalent to histology diagnosis are extremely valuable in oral cancer screening and early detection in resource-constrained settings. Optical coherence tomography (OCT), an optical biopsy technique enables real-time imaging with periodic surveillance and capability to image architectural features of the tissues. We report that while OCT system delineates oral pre-cancer and cancer with more than 90% sensitivity, integration, with artificial neural network-based analysis efficiently identifies high-risk, oral pre-cancer (83%). This study provides evidence that the robust, low-cost system was effective as a point-of-care device in resource-constrained settings. The high accuracy and portability signify widespread clinical application in oral cancer screening and/or surveillance.Non-invasive strategies that can identify oral malignant and dysplastic oral potentially-malignant lesions (OPML) are necessary in cancer screening and long-term surveillance. Optical coherence tomography (OCT) can be a rapid, real time and non-invasive imaging method for frequent patient surveillance. Here, we report the validation of a portable, robust OCT device in 232 patients (lesions: 347) in different clinical settings. The device deployed with algorithm-based automated diagnosis, showed efficacy in delineation of oral benign and normal (n = 151), OPML (n = 121), and malignant lesions (n = 75) in community and tertiary care settings. This study showed that OCT images analyzed by automated image processing algorithm could distinguish the dysplastic-OPML and malignant lesions with a sensitivity of 95% and 93%, respectively. Furthermore, we explored the ability of multiple (n = 14) artificial neural network (ANN) based feature extraction techniques for delineation high grade-OPML (moderate/severe dysplasia). The support vector machine (SVM) model built over ANN, delineated high-grade dysplasia with sensitivity of 83%, which in turn, can be employed to triage patients for tertiary care. The study provides evidence towards the utility of the robust and low-cost OCT instrument as a point-of-care device in resource-constrained settings and the potential clinical application of device in screening and surveillance of oral cancer.

Abstract 2539: DNA content and chromatin texture measurement to predict malignant transformation in low-grade oral dysplasia

Abstract Objectives: The key to reducing oral cancer morbidity and mortality is the early identification of oral potentially malignant lesions (OPMLs) at high risk of malignant transformation (MT). The gold standard to assess OPML risk is the grade of dysplasia. Risk prediction of low-grade dysplasias (LGDs) is challenging as only a small proportion progress. Currently no tools or biomarkers are used clinically to predict MT. Abnormal DNA content has shown to be a marker of malignancy in various sites including the oral cavity. Changes in nuclear morphology and chromatin texture have also been observed in cells with normal DNA content adjacent to cancerous fields as a result of genetic or epigenetic alterations that drive MT. The aim of this study is to use OPML lesion brushings to measure nuclear DNA content and morphometric features to identify oral LGDs at high risk of MT. Methods: Patients with oral LGDs consented into the British Columbia Oral Cancer Prediction Longitudinal Study were included in the study. Inclusion criteria included patients with mild or moderate dysplasia with lesion brushings prior to or concurrent to biopsy dates and had more than 6 months of follow up. Outcome was defined as progression to severe dysplasia or cancer. Thin monolayer slides were prepared and stained using Feulgen Thionin. Slides were scanned using the Cancer Imaging Scanner at BC Cancer using machine learning algorithms to measure DNA amount and over 120 nuclear morphometric features. Linear discriminant function (LDF) analysis of nuclear features from normal diploid cells were used to train classifiers to differentiate progressing from non-progressing lesions. Results: A total of 143 lesion brushings from 140 patients were selected. Forty-nine mild, 48 moderate, and 46 lesions with a previous history of dysplasia at the same site were included of which 16 (11%) progressed. DNA content of each nucleus was measured by a normalized scale known as DNA Index (DI) and were classified into groups: diploid (0.9 <DI<1.1), tetraploid (1.8<DI<2.2), proliferating (1.2<DI<1.7) and aneuploid (DI>2.5). Abnormal DNA content threshold was determined using combinations of cell frequencies/percentage of each group. A lesion was considered at high-risk of MT when it showed 5 or more non-diploid cells (DI>1.2). 12 out of 16 progressors and 41 out of 116 non-progressors showed high-risk DNA content. Preliminary results from LDF show that chromatin texture features best differentiated progressing cells from non-progressing. Analysis to determine the best threshold for chromatin texture features is ongoing. Conclusion: This suggests that chromatin texture changes can be observed in cells with normal DNA content during MT. Together, they can serve as a quick, non-invasive, cost effective tool to triage high risk OPMLs to cancer care centres for monitoring and early intervention. A larger sample size is required to validate our results. Citation Format: Madhurima Datta, Denise M. Laronde, Miriam Rosin, Anita Carraro, Korbelik Jagoda, Alan Harrison, Zhaoyang Chen, Martial Guillaud. DNA content and chromatin texture measurement to predict malignant transformation in low-grade oral dysplasia [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 2539.

Abstract 2813: A multiplexed, multispectral approach to analyzing the immune microenvironment of oral potentially malignant lesions

Abstract Objectives: Tissue sections of oral potentially malignant lesions (OPML) can be used not only to provide pathological assessment (diagnosis) but could also be used to analyze the interactions between cellular populations, signaling molecules, and structural proteins that impact the clinical course of disease. However, much of this information remains unpacked due to methodological limitations. Traditional immunohistochemistry (IHC) limits the number of proteins able to be simultaneously analyzed within a tissue section and is also prone to human error in its analysis. Newer multiplexed IHC (mIHC) methods involving repeated cycles of staining allow for quantification of a greater number of markers; however, tissue integrity may be compromised, and complexity is added to the interpretation of the results. There is a need to develop an immunostaining method that overcomes these barriers. Hypothesis: mIHC and an in-house Hyperspectral Cell Sociology (HCS) platform will allow for robust and detailed investigation of the immune microenvironment of OPML compared to traditional IHC staining and scoring techniques. Methods: Automated mIHC staining with a seven immune marker panel was completed on annotated formalin-fixed paraffin-embedded OPML tissue. A cocktail of three primary antibodies was applied, then antigens and chromogens stripped using SDS-glycine before second round staining with four antibodies and a hematoxylin counterstain was completed. Slides were then digitally imaged, regions of interest selected in conjunction with an oral pathologist and staining quantified computationally using the HCS platform. Traditional IHC was completed on a randomized subset of cases using sequentially cut tissue sections and a double-staining technique. Scoring was completed by two blinded clinicians. Results: One cycle of multiplexed staining and de-staining allowed for the detection of seven markers on one section while minimizing loss of tissue integrity. The HCS platform captures a single tissue section at multiple wavelengths, enabling the unmixing of multiple overlapping, colocalized chromogens. The resulting set of images displayed each stain separately, allowing for nuclei segmentation and the generation of a map of the epithelium and underlying connective tissue. True positive cells for each stain were demarcated on this map, allowing for investigation of marker positivity, co-positivity, cell to cell spatial relationships, and layer-based analysis, compared to cell count and density analyses obtained with traditional IHC. Conclusion: The immune microenvironment contains a wealth of information pertaining to the biology, pathogenesis, and outcome of disease. Multiplexed staining and imaging methods to analyze and unpack this information is of great clinical utility. Citation Format: Iris Lin, Kouther Noureddine, Paul Gallagher, Martial Guillaud, Lewei Zhang, Leigha Rock, Miriam Rosin, Denise Laronde. A multiplexed, multispectral approach to analyzing the immune microenvironment of oral potentially malignant lesions [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 2813.

Patients’ views on a proposed oral cancer screening technology

Brush cytology and telemedicine platforms are used with proven benefits in remote diagnosis of malignancies like cervical cancer. In low-resource settings access to the current standard of care, biopsy and histopathology grading of oral lesions, can be unfeasible. Risk-stratifying early malignant and oral potentially-malignant lesions can help instigate therapeutic treatment and improve prognosis. Therefore, a telecytology platform enabling remote connectivity to specialist centres may aid early detection of oral cancer. We outline head and neck (H&N) cancer patients’ views on an oral brush cytology investigation.

TISSUE FLUORESCENCE IMAGING FOR QUICK NON-INVASIVE DIAGNOSIS IN ORAL AND MAXILLOFACIAL SURGERY

Significant vital functions take place in the oral cavity and oropharynx, primarily mastication, as the initial function of the gastrointestinal system, swallowing, respiration and speech. All these vital biological functions can be endangered, ie aggravated, and even disabled by the appearance of neoplasms in that anatomical space. In the maxillofacial region, neoplasms can originate from a variety of tissues, from the mucous membranes of the oral cavity to the jaws, salivary glands, and even tumors of odontogenic origin. However, the most common tumor in the oral cavity is squamous cell carcinoma (OSSC), which originates from the oral mucosa. To determine the accuracy, sensitivity (Se) and specificity (Sp) of the Velscope screening method in the detection of premalignant and initial malignant lesions compared to the gold standard surgical biopsy. The study group consisted of 60 patients divided into two groups. The first group was formed by 30 patients with potentially malignant oral lesions (PML). Another 30 patients with preliminary diagnosis - oral cancer (OC) were included in the second group of examinees. The high sensitivity value of 92.86% and the accuracy of the method of 86.67% largely confirm the reliability and efficacy of the Velscope method in patients with highly suspected oral cancer lesions (OC), significantly more, comparing to the group of premalignant lesions. Key words: Velscope, oral cancer, premalignant lesions, tumor.&nbsp

The T102C polymorphism of 5HT2A receptor in oral epithelial dysplasia: A pilot case-control study

Objectiveinvestigate the T102C polymorphism of 5HT2A receptor in dysplasia in oral potentially malignant lesions and its association with smoking and alcohol habits. Methodscase-control study that included patients with oral potentially malignant lesions (OPML) histopathologically diagnosed with dysplasia and healthy controls, and within these group patients with and without smoking and alcohol consumption habits. Cell samples from the oral lesions were collected with the patients previously anesthetized using disposable cytological brushes. Deoxyribonucleic acid (DNA) extraction was performed and the T102C polymorphism (rs6313) was genotyped in a real-time polymerase chain reaction (PCR) allelic discrimination assays. Results110 individuals were included in this study (38 with dysplasia and 72 controls). The genotype (p = 0.016), allele (p = 0.020) and smoking habits (<0.001) distribution differed significantly between dysplasia and control group, where the CT and TT (C – cytosine/ T – thymine) genotype and the T allele showed a higher frequency in dysplasia (65.6, 18.8 and 84.4 %, respectively) than in controls (55.7, 4.9 and 60.7). Concerning smoking habits, the higher frequency was in the dysplasia group. The multivariate logistic regression analysis, associating variables of interest and the presence of dysplasia, showed that individuals with smoking habits present 7.58 increase risk to develop dysplasia than non-smokers; and individuals carrying the T allele for the T102C polymorphism have a 4.6 increased risk to develop oral dysplasia in OPML. Conclusionsthe T102C polymorphism is associated with oral dysplasia in OPML, however, failed to show association with smoking and alcohol habits in OPML dysplasia.

Point-of-care oral cytology tool for the screening and assessment of potentially malignant oral lesions.

BackgroundThe effective detection and monitoring of potentially malignant oral lesions (PMOL) are critical to identifying early‐stage cancer and improving outcomes. In the current study, the authors described cytopathology tools, including machine learning algorithms, clinical algorithms, and test reports developed to assist pathologists and clinicians with PMOL evaluation.MethodsData were acquired from a multisite clinical validation study of 999 subjects with PMOLs and oral squamous cell carcinoma (OSCC) using a cytology‐on‐a‐chip approach. A machine learning model was trained to recognize and quantify the distributions of 4 cell phenotypes. A least absolute shrinkage and selection operator (lasso) logistic regression model was trained to distinguish PMOLs and cancer across a spectrum of histopathologic diagnoses ranging from benign, to increasing grades of oral epithelial dysplasia (OED), to OSCC using demographics, lesion characteristics, and cell phenotypes. Cytopathology software was developed to assist pathologists in reviewing brush cytology test results, including high‐content cell analyses, data visualization tools, and results reporting.ResultsCell phenotypes were determined accurately through an automated cytological assay and machine learning approach (99.3% accuracy). Significant differences in cell phenotype distributions across diagnostic categories were found in 3 phenotypes (type 1 [“mature squamous”], type 2 [“small round”], and type 3 [“leukocytes”]). The clinical algorithms resulted in acceptable performance characteristics (area under the curve of 0.81 for benign vs mild dysplasia and 0.95 for benign vs malignancy).ConclusionsThese new cytopathology tools represent a practical solution for rapid PMOL assessment, with the potential to facilitate screening and longitudinal monitoring in primary, secondary, and tertiary clinical care settings.

Knowledge assessment among the fourth and fifth year students on the potentially malignant lesions of oral cavity

Background/Aim: Potentially malignant oral lesions (PMOL) are lesions that have an increased risk of malignant transformation concerning healthy oral mucosa. This research aimed to assess the knowledge of students of final years of study on the potentially malignant lesions of the oral cavity. Material and Methods: 120 students of the fourth and fifth years of study participated in this research,60 respondents were fourth-year students, 60 respondents were fifth-year students. All of the respondents signed informed consent.According to the type of research, this is a cross-sectional study which was conducted by completing a 15-item questionnaire. The results were statistically analyzed and processed in the SPSS Statistics 21.0 program. Results: The results indicate that during the clinical examination the fourth and fifth-year students examine the oral mucosa as well. 61% of the students find themselves poorly informed on the PMOL. In the fourth year of study, only 5,5% of students responded that they were well informed, while in the fifth year that number totaled 28,5%. When knowledge self-assessment is concerned the difference has shown itself to be statistically significant between the fourth and fifth-year students. When asked what knowledge on the prevention of oral cancer they expect to receive during their studies, 78% of the students expected more knowledge than at that moment - 36% of which were fifth-year students, and 42% were fourth-year students. Conclusions: Even though the oral cavity is easily accessible to examination, the discovery of oral cancer occurs quite late. Ours, as well as results of other studies, impose the need for better theoretical and practical education of the students.

Abstract 4223: Conducting community oral cancer screening among South Asians in British Columbia

Abstract Introduction: Globally, more than 300,000 cases of oral cancer are diagnosed annually. South Asian countries, such as India, bear the brunt of this disease due to rampant use of chewing tobacco, betel quid and areca nut. BC has a high proportion of South Asian immigrants. Oral cancer has a high mortality rate (~50% 5-year survival) due to the advanced stage at which it is often diagnosed. It is purported that the majority of oral cancers develop from oral potentially malignant lesions (OPML). While lesions can be easily detected by oral health care providers, it can be challenging to differentiate benign lesions from OPML. DNA aneuploidy has been shown to be an effective marker to predict malignant transformation in OPML. Quantitative cytology (QC) studies DNA content (ploidy) and nuclear morphometric changes within the cell. The aim of this project is to assess the need for oral cancer screening in South Asians in BC and to validate QC as an adjunct screening device in a predominantly South Asian community screening setting to assess its effectiveness in identifying high-risk lesions among visually suspicious lesions. Methods: Demographic information (gender, age, country of birth, ethnicity, risk habit information and dental usage) were collected at the time of screening. Extraoral, intraoral and fluorescence visualization (FV) examinations were conducted. Buccal mucosal brushings were collected from each participant. Brushings were also collected from lesions or areas that had a loss of fluorescence. Thin-layer cytology slides were prepared and stained using Feulgen Eosin. Slides were scanned using the Cancer Imaging Scanner at BC Cancer using machine learning classification algorithms to identify single, in-focus epithelial nucleus. Cells are classified based on DNA ploidy and malignant associated changes. Results: 307 participants were screened of which 303 were eligible. More than 99% of the participants were South Asian or Asian. 104 (34%) lesions were documented: 45 (15%) were high risk (white or red lesions, lichen planus (LP)) and 59 (19%) were low risk (trauma, candidiasis, aphthous ulcers). Twenty participants (7%) were suspected to have high-risk OPMLs (not LP): 12 were referred directly to our Next Gen Oral Dysplasia clinic for biopsy, while 8 were reassessed at 3 weeks. Chewing tobacco was found to be strongly associated with lesion presence (p&amp;lt;0.01). QC analysis is ongoing for 320 samples. To date, 5 biopsies have been performed resulting in 1 mild dysplasia, 1 severe dysplasia and 3 hyperplasia. Conclusion: South Asians in BC were found to be at high risk for OPMLs. QC may help to improve the sensitivity and specificity of oral cancer screening by distinguishing false FV positive inflammatory lesions from high-risk lesions. Citation Format: Leigha D. Rock, Madhurima Datta, Denise M. Laronde, Anita Carraro, Jagoda Korbelik, Alan Harrison, Martial Guillaud. Conducting community oral cancer screening among South Asians in British Columbia [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 4223.

Abstract 659: Expression patterns of NF-κB in inflammatory oral potentially malignant lesions

Abstract Introduction: Oral lichen planus (OLP), a common chronic autoimmune inflammatory condition, is recognized as a potentially malignant condition by the World Health Organization. However, some argue that only OLP with epithelial dysplasia - termed lichenoid dysplasia (LD) - have malignant potential. As research continues to characterize the immune microenvironment of OLP, there is a need to elucidate factors favorable for malignant change. Recent research has demonstrated that activation of nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) is key to cancer development. This transcription factor has been extensively explored in inflammation-associated cancers, such as colon and gastric cancers. There is a need to investigate these factors in oral potentially malignant lesions (OPML), including OPL. In this study, we aim to compare clinical and risk habit differences between OLP and LD, and determine if NF-κB expression is associated with histological and clinical features of OPML indicative of cancer risk. Methods: Clinical, demographic, and histological data have been collected from the Oral Cancer Prediction Longitudinal (OCPL) study and the CoPath Vancouver Coastal Health Database. Patients with a primary diagnosis of OLP or low-grade LD were eligible to participate. Patients with previous history of head and neck cancer, or who have less than one year of follow-up are excluded from enrollment. Demographic, risk habit and clinical information was collected.For completed cases, immunohistochemistry (IHC) has been performed on formalin-fixed and paraffin-embedded tissue. Nuclear reactivity of NF-κB in the epithelium was counted in 10 high-power fields, and cytoplasmic positivity classified into 4 categories. Chi-squared tests were performed on categorical demographic and risk habit data. Results: To date, 51 participants have been recruited into this ongoing study: 37 with OLP and 14 cases of LD. There is no significant difference in gender and age between groups (p=0.297, p=0.120, respectively). Ever smokers and lesion location at a high-risk site were significantly associated with a diagnosis of LD compared to OLP (p=0.002, p&amp;lt;0.001, respectively). Preliminary IHC results show strong NF-κB positivity in the sub-epithelial lymphocytic infiltrate and strong to moderate cytoplasmic reactivity in the epithelium. Nuclear reactivity appears to increase from lichen planus to dysplasia. Conclusion: Patients with LD were more apt to be smokers, and more often presented with lesions at a high-risk site compared to those with OLP. Strong NF-κB cytoplasmic positivity in OLP, especially adjacent to areas with intense cytotoxic inflammatory infiltrate, reinforces the prominent role of NF-κB in inflammation. Citation Format: Iris Lin, Lewei Zhang, Miriam Rosin, Leigha Rock, Denise Laronde. Expression patterns of NF-κB in inflammatory oral potentially malignant lesions [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 659.

EXPRESSSION OF SERUM AND SALIVARY MMP-1 AND MMP-9 IN ORAL CANCER, ORAL POTENTIALLY MALIGNANT DISORDERS AND NORMALS.

Background: Matrix metalloproteinases (MMPs) have been implicated in aggressiveness, invasiveness and metastasis of many malignancies including oral squamous cell carcinoma. In the meta-analysis of microarray based Oral Tongue SCC gene expression profiles, MMP-1 and MMP-9 was highly up-regulated .Till date, non-invasive diagnostic aid to detect the malignant potential of OPML or the aggressiveness of OSCC is lacking. With this background we decided to see the level of MMP-1 and MMP- 9 in saliva and serum of OSCC patients with controls.&#x0D; Aim: To determine the level of MMP1 and MMP9 in serum and saliva of patients diagnosed with OSCC, OPML, normal and to correlate with the aggressiveness of the disease.&#x0D; Methodology: The epidemiologic data of the patients reporting to our dental college &amp; Hospital were obtained through an interview. After obtaining written consent from the patient, complete intra oral examination was done, blood, saliva was collected and stored using the standard protocol. ELISA was done on all the samples for MMP-1 and MMP9.&#x0D; Results: MMP-1 in serum and MMP-9 in saliva/ serum was up-regulated in OSCC, moderately up-regulated in OPML when compared to the control group. However MMP-1 in saliva was found to be very low or non detectable.&#x0D; Conclusion: The MMP-9 in saliva/serum and MMP-1 in serum was up-regulated in the OSCC. In future, may be these body fluids can be used to diagnose and to determine aggressiveness of OPML and OSCC by non-invasive techniques.&#x0D; Keywords:Enzyme-Linked Immunosorbent Assay (ELISA ), Matrix Metalloproteinase (MMP),Oral Potentially Malignant Lesions (OPML), Oral Squamous Cell Carcinoma( OSCC), Oral Tongue Squamous Cell Carcinoma&#x0D; ( OTSCC).