High-grade Epithelial Dysplasia Research Articles

Immunophenotypic and Gene Expression Analyses of the Inflammatory Microenvironment in High-Grade Oral Epithelial Dysplasia and Oral Lichen Planus

BackgroundOral lichen planus (OLP) and oral epithelial dysplasia (OED) present diagnostic challenges due to clinical and histologic overlap. This study explores the immune microenvironment in OED, hypothesizing that immune signatures could aid in diagnostic differentiation and predict malignant transformation.MethodsTissue samples from OED and OLP cases were analyzed using immunofluorescence/immunohistochemistry (IF/IHC) for CD4, CD8, CD163/STAT1, and PD-1/PDL-1 expression. RNA-sequencing was performed on the samples, and data was subjected to CIBERSORTx analysis for immune cell composition. Gene Ontology analysis on the immune differentially expressed genes was also conducted.ResultsIn OED, CD8 + T-cells infiltrated dysplastic epithelium, correlating with dysplasia severity. CD4 + lymphocytes increased in the basal layer. STAT1/CD163 + macrophages correlated with CD4 + intraepithelial distribution. PD-1/PDL-1 expression varied. IF/IHC analysis revealed differential immune cell composition between OED and OLP. RNA-sequencing identified upregulated genes associated with cytotoxic response and immunosurveillance in OED. Downregulated genes were linked to signaling, immune cell recruitment, and tumor suppression.ConclusionsThe immune microenvironment distinguishes OED and OLP, suggesting diagnostic potential. Upregulated genes indicate cytotoxic immune response in OED. Downregulation of TRADD, CX3CL1, and ILI24 implies dysregulation in TNFR1 signaling, immune recruitment, and tumor suppression. This study contributes to the foundation for understanding immune interactions in OED and OLP, offering insights into future objective diagnostic avenues.

Setd2 deficiency promotes gastric tumorigenesis through inhibiting the SIRT1/FOXO pathway

Gastric cancer (GC) is the fifth most common cancer and the second leading cause of cancer death globally. SETD2 is a histone methyltransferase catalyzing tri-methylation of H3K36 (H3K36me3) and has been shown to participate in diverse biological processes and human tumors. However, the mechanism of SETD2 in GC remains unclear. Here, we reported that Setd2 deficiency predicts poor prognosis of gastric cancer. SETD2 loss facilitated H. felis/MNU and c-Myc-induced gastric tumorigenesis, respectively. The mouse model of stomach-specific Setd2 depletion together with c-MYC overexpression (AMS) developed high-grade epithelial defects, intestinal metaplasia and dysplasia at only 10–12 weeks of age. Mechanistically, Setd2 depletion resulted in impaired epigenetic regulation of Sirt1, thus inhibiting the SIRT1/FOXO pathway. Moreover, the agonists of FOXO signaling or overexpression of SIRT1 significantly rescued the enhanced cell proliferation and migration caused by Setd2 deficiency in SGC7901 cells. Together, our findings highlight an epigenetic mechanism by which SETD2 regulates gastric tumorigenesis through SIRT1/FOXO pathway. It may also pave the way for the development of targeted, patient-tailored therapies for GC patients with Setd2 deficiency.

A Novel Role for the Soluble Isoform of CTLA-4 in Normal, Dysplastic and Neoplastic Oral and Oropharyngeal Epithelia

Background: Head and neck cancer (HNC) has a high mortality rate, with late diagnosis remaining the most important factor affecting patient survival. Therefore, it is imperative to identify markers that aid in early detection and prediction of disease progression. HNCs evade the immune system by different mechanisms, including immune checkpoints. Cytotoxic T-lymphocyte-associated protein-4 (CTLA-4) is an immune checkpoint receptor that downregulates anti-tumour immune responses, with evidence of involvement in HNC. The less studied, alternatively spliced, soluble isoform (sCTLA-4) also plays an immunosuppressive role that contributes to immune escape. We quantified sCTLA-4 in normal, potentially malignant, and malignant oral and oropharyngeal tissues to elucidate any role in tumourigenesis and identify its potential as a biomarker for diagnosis and patient stratification. Methods: Normal, low- and high-grade epithelial dysplasia, and squamous cell carcinoma oral and oropharyngeal biopsies were selectively stained for sCTLA-4 and quantified using the image analysis software QuPath. Results: Distinct sCTLA-4 staining patterns were observed, in which normal epithelial sCTLA-4 expression correlated with keratinocyte differentiation, while disrupted expression, both in intensity and localisation, was observed in dysplastic and neoplastic tissues. Conclusions: Our data indicate an additional, previously unknown role for sCTLA-4 in epithelial cell differentiation and proliferation. Furthermore, our findings suggest the potential of sCTLA-4 as a biomarker for predicting disease progression and patient stratification for targeted HNC therapies.

Молекулярно-генетические критерии малигнизации эпителия слизистой оболочки рта

Introduction. When examining the oral mucosa (OM) in the presence of an inflammatory process, the initial signs of malignancy are not always visible. Amid an inflammatory infiltrate, benign changes such as epithelial hyperplasia (EH), epithelial dysplasia (ED) of varying severity, and squamous cell carcinoma (SCC) can be hidden. The aim of the paper was to study the proliferative activity of epithelial cells of the OM, TERT expression, and amplification of telomerase TERC gene in the development of epithelial hyperplasia, dysplasia, and SCC. Materials and methods. We divided 60 patients into four groups: Group 1 with EH (n=16, 26.6%); Group 2 with low-grade epithelial dysplasia (LGED) [n=16, 26.6%]; Group 3 with high-grade epithelial dysplasia (HGED) [n=13, 21.6%]; and Group 4 with SCC (n=15, 25%). IHC and histological studies were carried out using standard methods; expression to Ki-67 protein and telomerase activity were evaluated. The FISH method was used with LSP TERC and CCP3 DNA probes. Results. High-cell proliferation and telomerase activity were revealed by IHC study in all groups. Amplification of TERC RNA site in chromosome 3 by FISH in Group 1 was almost absent and showed a 2:2 signal ratio in 1.3% of cases; in other groups, the amplification of TERC gene was positive: in groups 2, 3, and 4 it was in 83.9%, 97.3%, and 89.8% of cases, respectively. Conclusion. Amplification of TERC site signals in chromosome 3 is detected during malignization which may be the main criterion for early diagnosis of malignization of OM epithelium. Keywords: squamous cell carcinoma, epithelial dysplasia, TERC amplification, telomerase

Whole-exome sequencing of oral epithelial dysplasia samples reveals an association with new genes.

The genetic basis of oral epithelial (OED) is unknown, and there is no reliable method for evaluating the risk of malignant transformation. Somatic mutations are responsible for the transformation of dysplastic mucosa to invasive cancer. In addition, these genomic variations could represent objective markers of the potential for malignant transformation. We performed whole-exome sequencing of 10 OED samples from Brazilian and Chilean patients. Using public genetic repositories, we identified 41 deleterious variants that could produce high-impact changes in the amino acid structures of 38 genes. In addition, the variants were filtered according to normal skin and Native American genome profiles. Finally, 13 genes harboring 15 variants were found to be exclusively related to OED. High-grade epithelial dysplasia samples showed a tendency to accumulate highly deleterious variants. We observed that 62% of 13 OED genes identified in our study were also found in head and neck squamous cell carcinoma. Among the shared genes, eight were not identified in oral squamous cell carcinoma. To our knowledge, we have described for the first time 13 genes that are found in OED in a Latin American population, of which five genes have already been observed in oral squamous cell carcinoma. Through this study, we identified genes that may be related to basal biological functions in OED.

Laminin Immunostaining in Biopsies as a Useful Biomarker of Early Invasion in Actinic Cheilitis and Differential Diagnosis Between Actinic Cheilitis and Lip Cancer: New Insights

BackgroundSquamous cell carcinoma of the lip (LSCC) and oral cavity can be life-threatening if not diagnosed early. Precancerous lesions like actinic cheilitis (AC), can transform into LSCC. Laminin is a fundamental component for basement membrane (BM) and its integrity may prevent neoplastic invasion. Therefore, laminin immunostaining of BM may be useful in identifying early invasion in actinic cheilitis and thus in the differential diagnosis between AC and invasive LSCC or high-grade epithelial dysplasia (ED).Materials and methodsBiopsies from 46 patients with oral lesions were histologically analyzed and immunohistochemically stained for laminin-1.ResultsAC was diagnosed in 34 patients and LSCC in 12 patients, including 3 patients with AC and concomitant high-grade ED/in situ carcinoma. Laminin-1 immunostaining revealed intense and linear expression of the BM in AC with low-grade ED. Loss of laminin expression was observed in LSCC. Intracellular laminin expression in parabasal cells was noted in AC with high-grade ED/in situ carcinoma.ConclusionLaminin immunostaining could be useful in identifying AC cases suspected of early invasion. It could also contribute to the histopathological differential diagnosis between AC with low- and high-grade ED and between AC and invasive LSCC. The findings of this study provide new insights into the mechanism involved in the progression process of AC into LSCC, encouraging preclinical studies that may document the stochastic role of laminin in this process.

Living guideline on ulcerative colitis

The treatment spectrum for ulcerative colitis has greatly increased in recent years. Mesalazine is still the standard drug treatment for uncomplicated ulcerative colitis in various forms of administration. Glucocorticoids are highly effective in the acute treatment of ulcerative colitis but should only be used on a short-term basis due to the pronounced side effects. For forms with a complicated course of ulcerative colitis, immunosuppressive and immunomodulating substances, such as azothioprine as well as various biologicals, Janus kinase inhibitors (JAKi), sphingosine-1-phosphate receptor agonists (S1PR agonists) and calcineurin inhibitors are available after failure of conventional treatment. A proctocolectomy should be considered in cases of a treatment-refractive course or with detection of carcinomas and high-grade epithelial dysplasia.

Melanoma Inhibitory Activity and Melanoma Inhibitory Activity 2 as Novel Immunohistochemical Markers of Oral Epithelial Dysplasia.

Oral potentially malignant disorders are associated with the development of oral squamous cell carcinoma (OSCC). Most OSCCs are diagnosed via histopathology as oral epithelial dysplasia (OED), but the histologic diagnostic criteria remain non-uniform. Accordingly, the establishment of a diagnostic marker to assist in diagnosis could contribute towards cancer prevention. Melanoma inhibitory activity (MIA) and MIA2 are involved in tumor growth, invasion, and lymph node metastasis in various malignancies. The purpose of this study was to clarify the usefulness of MIA and MIA2 as diagnostic markers of oral mucosal lesions. The expression of MIA and MIA2 was analyzed immunohistochemically in 100 specimens (10 specimens with normal oral mucosa (NOM) and 30 specimens each with low-grade epithelial dysplasia (LED), high-grade epithelial dysplasia (HED), and OSCC). Immunohistochemical results were evaluated based on the Allred scoring system. Cytoplasmic expression of MIA and MIA2 increased in the order of LED, HED, and OSCC. All NOM specimens were negative for cytoplasmic expression. Significant differences were observed between the groups (NOM vs. HED, p < 0.05, NOM vs. OSCC, p < 0.001). These results demonstrate that MIA and MIA2 are expressed in the oral mucosa within early neoplastic lesions and suggest that MIA and MIA2 are useful novel immunohistochemical markers for discriminating between normal tissue and OED.

CONSERVATIVE APPROACH FOR ACTINIC CHEILITIS WITH HIGH-GRADE EPITHELIAL DYSPLASIA

Actinic cheilitis (AC) is a potentially malignant disorder of the lip caused by chronic exposure to ultraviolet radiation. This study aimed to report a case of AC with high-grade epithelial dysplasia submitted to a conservative treatment. Patient E.F.N.F., a 50-year-old man with melanoderma, sought dental care because of the presence of a lesion in the lower lip with a history of about 6 months. He denied painful symptoms. On clinical examination, fissures, edema, and presence of nonraised ulcer showing reddish central area and leucoplast peripheral region were observed in the lower lip. The patient was advised to undergo diagnostic biopsy. The anatomopathologic report resulted in AC with high-grade epithelial dysplasia. He was advised to use hats in addition to moisturizer and sunscreen. After 2 months significant regression of the ulcer and improvement of lip edema was observed. Currently, the patient is in observation, and reviews are made every 3 months. Actinic cheilitis (AC) is a potentially malignant disorder of the lip caused by chronic exposure to ultraviolet radiation. This study aimed to report a case of AC with high-grade epithelial dysplasia submitted to a conservative treatment. Patient E.F.N.F., a 50-year-old man with melanoderma, sought dental care because of the presence of a lesion in the lower lip with a history of about 6 months. He denied painful symptoms. On clinical examination, fissures, edema, and presence of nonraised ulcer showing reddish central area and leucoplast peripheral region were observed in the lower lip. The patient was advised to undergo diagnostic biopsy. The anatomopathologic report resulted in AC with high-grade epithelial dysplasia. He was advised to use hats in addition to moisturizer and sunscreen. After 2 months significant regression of the ulcer and improvement of lip edema was observed. Currently, the patient is in observation, and reviews are made every 3 months.

Ulcerative Colitis-Diagnostic and Therapeutic Algorithms.

Ulcerative colitis is a chronic inflammatory bowel disease with an estimated 150 000 patients in Germany alone. This review is based on publications about current diagnostic and therapeutic strategies for ulcerative colitis that were retrieved by a selective search in PubMed, and on current guidelines. The primary goal of treatment is endoscopically confirmed healing of the mucosa. Mesalamine, in various forms of administration, remains the standard treatment for uncomplicated ulcerative colitis. Its superiority over placebo has been confirmed in meta-analyses of randomized, controlled trials. Glucocorticoids are highly effective in the acute treatment of ulcerative colitis, but they should only be used over the short term, because of their marked side effects. Further drugs are available to treat patients with a more complicated disease course of ulcerative colitis, including azathioprine, biological agents, JAK inhibitors (among them TNF antibodies, biosimilars, ustekinumab, vedolizumab, and tofacitinib), and calcineurin inhibitors. Proctocolectomy should be considered in refractory cases, or in the presence of high-grade epithelial dysplasia. Ulcerative colitis beginning in childhood or adolescence is often characterized by rapid progression and frequent comorbidities that make its treatment a special challenge. A wide variety of drugs are now available for the treatment of ulcerative colitis, enabling the individualized choice of the best treatment for each patient. Regular surveillance colonoscopies to rule out colon carcinoma should be scheduled at intervals that depend on risk stratification.

Diagnostic accuracy of brush biopsy-based cytology for the early detection of oral cancer and precursors in Fanconi anemia.

Individuals with Fanconi anemia (FA) have a 500-fold to 700-fold elevated risk, much earlier onset, and limited therapeutic options for oral squamous cell carcinoma (SCC) compared with the general population. The early detection of SCC, or preferably its precursors, is mandatory to retain curative therapeutic options. Due to frequent synchronic and metachronic oral lesions, tissue biopsies, as usually recommended by guidelines, often are not feasible. In the current study, an alternative strategy for early detection using oral brush biopsy-based cytology was validated regarding its diagnostic accuracy. Over a 12-year period, the oral cavities of a large cohort of 713 individuals with FA were inspected systematically and brush biopsy-based cytology of 1233 visible oral lesions was performed. In cases of inconclusive cytology, analysis of DNA ploidy was performed whenever possible. The results were correlated to a long-term clinicopathological follow-up reference standard. A total of 737 lesions were suitable for statistical analysis, including 86 lesions with at least high-grade oral epithelial dysplasia in 30 patients. For cytology, the sensitivity and specificity were 97.7% and 84.5%, respectively. Additional analysis of DNA ploidy increased the sensitivity and specificity to 100% and 92.2%, respectively. Careful inspection of the oral cavity of individuals with FA followed by brush biopsy-based cytology appears to identify visible oral, potentially malignant and malignant lesions that warrant treatment. Approximately 63% of SCC and precursor lesions are detected at a noninvasive or early stage. Negative cytology or a lack of DNA aneuploidy can exclude high-grade oral epithelial dysplasia or SCC with high accuracy and thus reduce the need for invasive diagnostic biopsies.

BMI-1 expression increases in oral leukoplakias and correlates with cell proliferation.

Oral leukoplakia (OL) is a white lesion of an indeterminate risk not related to any excluded (other) known diseases or disorders that carry no increased risk for cancer. Many biological markers have been used in an attempt to predict malignant transformation; however, no reliable markers have been established so far. Objective To evaluate cell proliferation and immortalization in OL, comparing non-dysplastic (Non-dys OL) and dysplastic OL (Dys OL).Methodology This is a cross-sectional observational study. Paraffin-embedded tissue blocks of 28 specimens of Non-dys OL, 33 of Dys OL, 9 of normal oral mucosa (NOM), 17 of inflammatory hyperplasia (IH), and 19 of oral squamous cell carcinomas (OSCC) were stained for Ki-67 and BMI-1 using immunohistochemistry.Results A gradual increase in BMI-1 and K-i67 expression was found in oral carcinogenesis. The immunolabeling for those markers was higher in OSCC when compared with the other groups (Kruskal-Wallis, p<0.05). Ki-67 expression percentage was higher in OL and in IH when compared with NOM (Kruskal-Wallis/Dunn, p<0.05). Increased expression of BMI-1 was also observed in OL when compared with NOM (Kruskal-Wallis/Dunn, p<0.05). No differences were observed in expression of both markers when non-dysplastic and dysplastic leukoplakias were compared. A significant positive correlation between Ki-67 and BMI-1 was found (Spearman correlation coefficient, R=0.26, p=0.01). High-grade epithelial dysplasia was associated with malignant transformation (Chi-squared, p=0.03).Conclusions These findings indicate that BMI-1 expression increases in early oral carcinogenesis and is possibly associated with the occurrence of dysplastic changes. Furthermore, our findings indicate that both Ki-67 and BMI-1 are directly correlated and play a role in initiation and progression of OSCC.

Assessing the Expression of Aquaporin 3 Antigen-Recognition Sites in Oral Squamous Cell Carcinoma.

Aquaporin 3 (AQP3) serves as a water and glycerol transporter facilitating epithelial cell hydration. Recently, the involvement of AQP3 in cancers has been reported. However, the immunohistochemical expression of AQP3 in carcinomas remains controversial. We hypothesized that differences in aquaporin 3 antigen recognition (AQP3 AR) may influence their expressions. Thus, our study aimed to assess the immunostaining patterns of 3 AQP3 AR sites in oral squamous cell carcinoma (OSCC) and to compare the adjacent areas of high-grade epithelial dysplasia (HG-ED) and normal oral mucosa (NOM). The study group included formalin-fixed OSCC samples (n=51) with adjacent regions of HG-ED (n=12) and NOM (n=51). The tissues were stained with anti-AQP3 antibodies (AR sites at amino acid (AA) 250-C terminus, AA180-228, and N terminus AA1-80) by immunohistochemistry. Our results showed that strong membranous immunostaining was observed for AQP3 AR sites at the AA250-C terminus and AA180-228 in all the samples for NOM and weak AQP3 immunostaining for both the AR sites in all the 12 samples for HG-ED. The invasive front of OSCC samples showed that AQP3 AR at the AA250-C terminus decreased in 42/51 samples (82.4%) and AA180-228 in 47/51 samples (92.2%). Conversely, in the AQP3 AR site at N terminus AA1-80, all samples of the NOM showed negative or slightly positive staining in the cytoplasm of the lower layers. AQP3 expression was increased in 12/12 cases (100%) and 46/51 cases (90.2%) in the HG-ED and invasive front of OSCC, respectively. AQP3 may be used as a biomarker for detecting malignant transformations. AQP3 AR site differences influence their immunohistochemical expression in OSCC.

Immunohistochemical assessment of growth factor signaling molecules: MAPK, Akt, and STAT3 pathways in oral epithelial precursor lesions and squamous cell carcinoma.

Several growth factors and their receptors, such as epidermal growth factor receptor, have been studied as prognostic biomarkers for many epithelial malignancies. The signal transduction cascade of those receptors includes RAS/RAF/ERK, PI3K/Akt/mTOR, and STAT3 pathways. The aim of this study was to investigate the expression levels of several key proteins of those pathways in patients with oral squamous cell carcinoma (OSCC) and oral epithelial precursor lesions (OEPLs), and to correlate the expressions of these proteins with clinicopathologic features and prognosis. Fifteen leukoplakia (LP), 15 low-grade epithelial dysplasia, 15 high-grade epithelial dysplasia (HD), and 132 OSCC specimens were immunohistochemically examined for KRAS, HRAS, NRAS, BRAF, pERK1/2, pAkt, pmTOR, and pSTAT3 expression. Immunoreactivity for these molecules predominantly occurred in regions OEPL basal to prickle layers and in most OSCC cells. KRAS and NRAS expression was significantly lower in OSCC than in OEPLs, while pAkt and pmTOR showed higher expression in OSCC than in OEPLs. pERK1/2 expression was significantly higher in HD than in LP. In OSCC, KRAS and NRAS immunoreactivity was significantly higher in advanced age and male gender. In addition, higher immunoreactivity was shown in pERK1/2 in female gender and advanced TNM stage, pAkt in advanced T classification and cases without postoperative metastasis, pmTOR in advanced mode of invasion, and pSTAT3 in invasion depth. Correlations between these markers and clinicopathological variables were also noted. MAPK, Akt, and STAT3 pathways might play diverse roles in oral carcinogenesis.

Expression of cornulin in oral premalignant lesions

The aim of this study was to evaluate expression of cornulin in oral mucosa as an adjunct to histopathologic grading of oral epithelial dysplasia (OED). Biomarker expression was assessed in normal oral mucosa, low-grade OED (LD), high-grade OED (HD), and oral squamous cell carcinoma (OSCC) by using immunohistochemistry. Photomicrographs were evaluated with Aperio Imagescope using a positive-pixel-counting algorithm. A histo-score (H-score) was calculated on the basis of staining intensity and the percentage of positive cells (%-staining). Intrarater reliability for H-score and %-staining was determined by calculating interclass correlation coefficients. Mean differences in H-scores and %-percent staining values were each analyzed by using an analysis of variance and Tukey's post hoc procedure. Cornulin expression progressively diminished with increasing grades of dysplasia and OSCC. Interclass correlation coefficients for H-score and %-staining were each greater than 0.99. Except for OSCC versus HD, all other pairwise comparisons were statistically significant (P < .0001) for H-score and %-staining. Cornulin expression helped differentiate between low-grade and high-grade oral epithelial dysplasia, making it a potential adjunct for grading oral OEDs and a potential biomarker for risk of lesion progression. Longitudinal studies evaluating risk stratification based on cornulin expression may be warranted.

Is the routine histopathological analysis of all gallbladders justified?

66,000 cholecystectomies are performed annually in the UK. Nationwide, all gallbladders are routinely sent for histological analysis to exclude incidental malignancy. This comes at significant cost to an overstretched National Health Service. This study set out to assess the utility of routine histopathological analysis following cholecystectomy. We performed a large, single-centre, retrospective analysis of all gallbladder specimens examined between February 2013 and December 2016 at our district general hospital. Specimens obtained in elective and emergency cases, through laparoscopic or open surgery, were included. A total of 752 gallbladder specimens were analysed. The mean age at the time of resection was 51 years, and the female:male ratio was 2.8:1. Seven (0.93%) specimens were reported to show suspicious histopathological features; epithelial atypia (1), low-grade epithelial dysplasia (2), high-grade epithelial dysplasia (1), papillary adenoma (1), MALT lymphoma (1) and invasive adenocarcinoma (1). In all seven cases, there was no clinical suspicion of gallbladder malignancy prior to surgery or suspicious pre-operative ultrasound scan findings. Gross macroscopic changes were noted at the time of resection in three cases. Four patients required further investigation and subsequent follow-up. All specimens deemed to be suspicious derived from patients over 50 years of age at the time of resection. Pre-operative investigations and macroscopic appearance at cholecystectomy may be inadequate in identifying patients with clinically important pathology. Routine histological analysis allows early identification. However, further work will assess whether more selective requests may help achieve the balance between cost-saving and safety in the context of a resource-limited healthcare service.

Viral DNA integration and methylation of human papillomavirus type 16 in high-grade oral epithelial dysplasia and head and neck squamous cell carcinoma.

This study evaluated the integration and methlyation of human papillomavirus type 16 (HPV16) in head and neck squamous cell carcinoma (HNSCC) and its oral precursor, high-grade oral epithelial dysplasia (hgOED). Archival samples of HPV16-positive hgOED (N = 19) and HNSCC (N = 15) were evaluated, along with three HNSCC (UMSCC-1, -47 and -104) and two cervical cancer (SiHa and CaSki) cell lines. HgOED cases were stratified into three groups with increasing degrees of cytologic changes (mitosis, karyorrhexis and apoptosis). The viral load was higher and the E2/E6 ratio lower (indicating a greater tendency toward viral integration) in group 3 than in groups 1 or 2 (p = 0.002, 0.03). Methylation was not observed in hgOED cases and occurred variably in only three HNSCC cases (26.67%, 60.0% and 93.3%). In HNSCC cell lines, lower E7 expression correlated with higher levels of methylation. HgOED with increased cytologic change, now termed HPV-associated oral epithelial dysplasia (HPV-OED), exhibited an increased viral load and a tendency toward DNA integration, suggesting a potentially increased risk for malignant transformation. More detailed characterization and clinical follow-up of HPV-OED patients is needed to determine whether HPV-OED is a true precursor to HPV-associated HNSCC and to clarify the involvement of HPV in HNSCC carcinogenesis.

A comparative immunohistochemical study of presence and distribution pattern of stromal myofibroblast in oral dysplasia and in different grades of oral squamous cell carcinoma

Aims and Objectives:Squamous cell carcinoma (SCC) is a common oral malignancy with a poor survival rate. Early tumorigenesis is marked by transdifferentiation of fibroblasts to myofibroblasts (MFs), which is supported by growth factors and cytokines expressed by tumor cells. The expression of alpha-smooth muscle actin (αSMA) marker correlates with the activation of MFs. This study was undertaken to compare the frequency and distribution of αSMA immunoexpression in oral epithelial dysplasia (OED) and OSCC.Materials and Methods:This study was conducted on samples collected from patients with oral epithelial dysplasia and oral SCC who visited Rajarajeswari Dental College and Hospital, Bengaluru. Tissue sections were subjected to Immunohistochemistry using αSMA marker, and cells were counted. The obtained data was subjected to Kruskal–Wallis test and Mann–Whitney U-test.Results:On performing Kruskal–Wallis test and Mann–Whitney U-test between the three groups (normal oral mucosa, OED, and OSCC) statistically significant result was found in the frequency between OED and OSCC and between normal tissue and OSCC. On comparing the distribution pattern, statistically significant result was found between OED and OSCC and between normal tissue and OSCC.Conclusion:The expression of MFs increases as the disease progresses from high-grade epithelial dysplasia to invasive OSCC. Poorly differentiated SCC showed more attendance of positive MFs in the stroma than other grades of OSCC. The rise in the number of αSMA-positive MFs and change in distribution pattern in OSCC can be associated with tumor invasive characteristics. Thus, the proliferation of MFs may be used as a stromal marker of premalignancy and malignancy.

Gallbladder polyps in familial adenomatous polyposis

Introduction Familiar adenomatous polyposis (FAP) is an inherited autosomal dominant disease characterized by the development of hundreds to thousands of colorectal adenomas in young adults. If left untreated, the patients develop colorectal cancer by age of 40. This disorder is caused by germline mutation of the adenomatous polyposis coli (APC) gene located on the chromosome 5q21. FAP can be associated with occur of various extracolonic malignancies, especially ampullary adenocarcinoma. As well it has been described an association between FAP, thyroid cancer and biliary system neoplasia. We present a rare case of multifocal gallbladder polyps in association with familial adenomatous polyposis. Case report We present the case of a 39-year-old male patient with the FAP-Syndrome. He had undergone a total colectomy with ileoanal pouch reconstruction years ago. In the current history, he reported about chronic abdominal pain in the right upper abdomen with postprandial nausea. The clinical examination at the admission showed no palpable mass or tenderness in the abdomen. The laboratory tests were normal. Abdominal ultrasound showed four concrements and multiple small polyps. He underwent laparoscopic cholecystectomy. The postoperative course was uneventful. Results and conclusions The mucosal inspection at the histopathological examination showed more than 80 green polypoid lesions. Microscopically were classified tubular adenomas with mostly low–grade but focally with high-grade epithelial dysplasia cells. Their epithelium was predominantly of the biliary type. No invasive malignant cells have been described. Immunohistochemical staining with s-catenin showed strong positivity for cytoplasmic and nuclear expression. APC is closely connected to s-catenin and a nuclear expression comes along with s-catenin mutations. Nuclear staining has been described in adenocarcinomas and late adenomas with severe dysplasia. Take-home message FAP is the most common adenomatous polyposis syndrome characterized by the development of numerous colorectal adenomatous polyps. It is associated with greater risk of developing upper gastrointestinal malignancy or extraintestinal cancer. Due to previous reports and our conclusions, a simultaneous prophylactic cholecystectomy should be performed by FAP patients, who undergo colectomy, to eliminate the risk of gallbladder neoplasia.

Programmed death ligand 1 (PD-L1) expression and tumor microenvironment: Implications for patients with oral precancerous lesions

ObjectivesCancer immunoediting represents a relatively novel concept attempting to explain the process of tumor escape from the host immune system response. Here, we attempted to elucidate the role of programmed death ligand 1 (PD-L1), the tumor microenvironment, and tumor escape mechanisms that allow malignant transformation of oral precancerous lesions. Materials and methodsPatients with oral precancerous lesions managed at the Nara Medical University Hospital, Japan, (n=120) were enrolled in this study. Epithelial dysplasias were graded by experienced pathologists, and subepithelial PD-L1-, CD163-, and CD8-positive cells were counted in the superficial lamina propria of oral mucosa. Epithelial PD-L1 expression was evaluated according to the staining intensity. The association of clinicopathological factors with epithelial dysplasia, malignant-free survival time, and significance of risk factors for malignant transformation were determined. ResultsMultivariate analysis showed that the subepithelial CD163-positive cell count was the only significant risk factor for high-grade epithelial dysplasia (P<0.001), while subepithelial CD163- and PD-L1-positive cell counts, and epithelial PD-L1 positivity were significantly associated with malignant-free survival (P=0.004, 0.04, and <0.001, respectively). Subepithelial PD-L1-positive cell count and epithelial PD-L1 positivity were significantly associated with malignant transformation (P=0.01 and 0.04, respectively). ConclusionOur results indicate that PD-L1-expressing dysplastic epithelial and recruited subepithelial cells in oral precancerous legions may evade the host immune system, and that the inhibition of PD-1/PD-L1 pathway may potentially prevent malignant transformation of oral precancerous legions as well as can treat advanced cancers.