Strong Staining Intensity Research Articles

FAP-positive activated fibroblasts are detectable in the microenvironment of endometriosis and correlate with stroma composition and infiltrating CD8+ and CD68+ cells

Abstract STUDY QUESTION Do activated fibroblasts expressing fibroblast activation protein-α (FAP)—which is traceable in positron emission topography/computed topography (PET/CT)—play a role in the microenvironment of endometriosis? SUMMARY ANSWER Activated fibroblasts expressing FAP are detectable in endometriotic lesions and correlate with iron and collagen content and infiltrating CD8-positive cytotoxic T cells and CD68-positive macrophages in the microenvironment endometriotic lesions. WHAT IS KNOWN ALREADY FAP-positive activated fibroblasts are found in various fibrosis-related pathologies and in the desmoplastic stroma of solid tumors; they can be traced in PET/CT but have not been investigated in the context of endometriosis, a chronic disease involving hormone-mediated repetitive tissue remodelling and fibrosis. STUDY DESIGN, SIZE, DURATION We analysed a cohort of endometriosis patients (n = 159) who had undergone surgery with removal of endometriotic foci at our University Hospital (tertiary care center) between 2018-2024. All patients provided written informed consent. The median age of the patients was 34 years. In total, 245 samples from different locations were analysed retrospectively. PARTICIPANTS/MATERIALS, SETTING, METHODS We investigated the expression of FAP and its relation to stroma composition and the immune microenvironment of endometriosis in 245 specimens from peritoneal lesions, ovarian endometriomas, deep infiltrating endometriosis and extra-abdominal lesions using conventional histology and immunohistochemistry followed by digital image analysis. Tissue within a radius of 500 µm of ectopic endometrium-like epithelium was analysed. To measure FAP expression in the perilesional stroma, a histoscore (H-score) was calculated. Masson trichrome staining was used to determine collagen content. Prussian blue staining for iron was used for age-dating of lesions. The abundance of CD68-positive macrophages and CD8-positive cytotoxic T cells within the microenvironment of ectopic endometriotic glands was analysed. Extra-lesional tissue served as controls. MAIN RESULTS AND THE ROLE OF CHANCE Distinct FAP expression (H-score > 10) was observed in 84% of endometriotic lesions and in only 4% of extra-lesional controls. FAP expression was significantly higher in endometriotic lesions (mean H-score of 61.8) than in extra-lesional tissue (mean H-score 3.8, p < 0.0001). There was a significant (p < 0.05) association with collagen content when comparing samples with low (H-score < 100) and high (H-score ≥ 100) FAP expression, and a significant difference in FAP expression correlating with the tissue iron content when comparing strong staining intensity and negative samples (p < 0.0005) or samples with weak staining intensity (p < 0.005). Moreover, the abundance of CD8+ and CD68+ cells was significantly higher (p < 0.0001) in samples with high FAP expression (H-score ≥ 100). LARGE SCALE DATA The data underlying this article will be shared upon reasonable request to the corresponding author. LIMITATIONS, REASONS FOR CAUTION This study proves the presence of FAP-positive fibroblasts in endometriosis by immunohistological methods. However, to translate targeting FAP into endometriosis diagnostics, these results have to be compared to imaging data and FAP inhibitor (FAPi) PET/CT has to be validated in a structured way on a large patient cohort. Moreover, we show that FAP expression is intertwined with the immune cell infiltrate in the microenvironment of endometriosis. To explore and to understand mechanisms contributing to chronic inflammation, immune evasion and fibrosis, more studies including more immune cell subtypes and functional experiments, are needed. WIDER IMPLICATIONS OF THE FINDINGS FAP-positive activated fibroblasts not only impact the immune microenvironment of endometriosis and are linked to increased macrophage and cytotoxic T cell infiltration, as we showed, but could also provide new options for non-invasive diagnostic methods and an improvement of the diagnostic workup prior to surgery. FAPi PET/CT should be considered when exploring new diagnostic options in endometriosis. STUDY FUNDING/COMPETING INTEREST(S) This work was funded by the Deutsche Forschungsgemeinschaft (DFG, German Research Foundation) “Clinician Scientist Program in Evolutionary Medicine” (project number 413490537 to FK). The authors declare that they have no conflicts of interest.

Tumor- and host-derived heparanase-2 (Hpa2) attenuates tumorigenicity: role of Hpa2 in macrophage polarization and BRD7 nuclear localization

Little attention was given to heparanase 2 (Hpa2) over the last two decades, possibly because it lacks a heparan sulfate (HS)-degrading activity typical of heparanase. Emerging results suggest, nonetheless, that Hpa2 plays a role in human pathologies, including cancer progression where it functions as a tumor suppressor. Here, we examined the role of Hpa2 in cervical carcinoma. We report that high levels of Hpa2 correlate with prolonged survival of cervical carcinoma patients. Strong staining intensity of Hpa2 also correlates with low tumor grade. Overexpression of Hpa2 in SiHa cervical carcinoma cells resulted in tumor xenografts that were two-fold smaller than control tumors. Interestingly, even smaller tumor xenografts were developed by SiHa cells overexpressing the Pro140Arg and Asn543Ile Hpa2 missense mutations that were identified in patients diagnosed with urofacial syndrome (UFS). Utilizing the Ras recruitment system, we identified bromodomain-containing protein 7 (BRD7) to interact with Hpa2 and found that both BRD7 and the Hpa2 mutants are translocated to the cell nucleus in tumors developed by the Pro140Arg and Asn543Ile Hpa2 mutants. Utilizing our newly developed conditional Hpa2-KO mice, we further show that Hpa2 plays a critical role in macrophage polarization; in the absence of Hpa2, macrophages are shifted towards pro-tumorigenic, M2 phenotype. Notably, implanting SiHa cervical carcinoma cells together with Hpa2-KO macrophages promoted tumor growth. These results support, and further expand, the notion that Hpa2 functions as a tumor suppressor, co-operating with another tumor suppressor, BRD7.

Immunohistochemical Expression of ROR2 in Gastrointestinal Stromal Tumor.

Gastrointestinal stromal tumors (GIST) are the most common mesenchymal tumors affecting the digestive tract, comprising approximately 0.1-3% of all gastrointestinal cancers. ROR2, a member of the receptor tyrosine kinase orphan receptor subfamily, functions as a signaling receptor for Wnt ligands. This study aims to assess the prognostic significance of ROR2 expression in GIST. A total of 56 paraffin-embedded blocks of GIST cases originating from different parts of the gastrointestinal tract (GIT) were included in this study. Immunohistochemistry was performed to detect ROR2 expression. ROR2 expression was observed in 71.4% of GIST cases, and strong intensity of ROR2 staining was significantly associated with prolonged overall survival of GIST patients (p = 0.048). Furthermore, ROR2 positivity and the percentage of immunostaining showed an inverse correlation with tumor progression. GIST cases displaying ROR2 positivity exhibit a reduced likelihood of disease progression and demonstrate favorable prognostic characteristics.

TRPS1 expression in MPNST is correlated with PRC2 inactivation and loss of H3K27me3

Initially described as a highly specific immunohistochemical marker for carcinomas of mammary origin, trichorhinophalangeal syndrome type 1 (TRPS1) has subsequently been detected in a variety of other non-mammary tumors. In this study, we examined the immunohistochemical expression of TRPS1 in 114 peripheral nerve sheath tumors, including 43 malignant peripheral nerve sheath tumors (MPNSTs), 58 schwannomas, including 9 cellular neurofibromas, and 13 neurofibromas, including 1 atypical neurofibroma. Notably, TRPS1 was expressed in 49% of MPNSTs and was absent in all schwannomas and neurofibromas. All MPNSTs showed TRPS1 labeling in >50% of nuclei, with 95% of cases demonstrating diffuse labeling. Most cases (67%) showed weak TRPS1 immunoreactivity, while a smaller subset showed moderate (24%) or strong (9%) intensity staining. Analysis of publicly available gene expression datasets revealed higher levels of TRPS1 mRNA in MPNSTs with PRC2 inactivation. In keeping with this finding, TRPS1 expression was more commonly observed in MPNSTs with loss of H3K27me3, suggesting a potential relationship between TRPS1 and the PRC2 complex. This study further broadens the spectrum of TRPS1-expressing tumors to include MPNST.

"GLI1 Subcellular Localization and Overexpression as Prognostic Factors for Disease-Free Survival in Colorectal Carcinoma".

Glioma-associated oncogene homolog-1 (GLI1) is amplified in human glioblastoma, and there is growing evidence suggesting its significant role in tumor development and metastasis. Our aim was to investigate the role of the GLI-1 gene in the progression of colorectal cancer (CRC) and its correlation with various clinicopathological features. Additionally, we examined the impact of the GLI-1 gene and other factors on the prognosis of CRC. We analyzed a total of 98 confirmed CRC cases and adjacent normal tissue controls. Patients suspected of having colon cancer underwent a colonoscopy and targeted biopsy, while those with rectal cancer underwent CT scans and MRI. GLI1 expression was detected using real-time PCR assay, Western blotting, and immunohistochemistry. The GLI1 gene was observed to be overexpressed in tumor tissues at both the protein and mRNA levels (p < 0.05). In addition, GLI1 overexpression was significantly associated with various factors such as tumor invasion (T3/T4), presence of lymph nodes, lymph node metastasis (LNM), stage (III/IV), tumor site (colon), tumor size (≥ 3cm), localization (nucleocytoplasmic), strong staining intensity and recurrence (p < 0.05). The results of survival analysis showed that the patients with overexpression of GLI1 had a significantly lower DFS rate which was 21months compared to those with normal expression who had 31months (p < 0.05). Moreover, individuals with early onset disease (15months) were more likely to have cytoplasmic localization of the GLI1 gene as opposed to nucleo-cytoplasmic localization of GLI1 which presented late-onset disease( 23months) (p < 0.05). Finally, Stage and PNI (p < 0.05) were found to independently affect outcomes of CRC according to Cox regression analysis. High expression of GLI-1 in CRC is associated with adverse pathology and poor prognosis for patients. The correlation between cytoplasmic localization of GLI-1 and reduced disease-free survival holds potential for guiding prognosis and treatment. Further research is needed to develop strategies targeting GLI-1 for improved outcomes.

Asparaginase-like protein 1 as a prognostic tissue biomarker in clinicopathologically and molecularly characterized endometrial cancer

ObjectivePrognostic stratification of endometrial cancer involves the assessment of stage, uterine risk factors, and molecular classification. This process can be further refined through annotation of prognostic biomarkers, notably L1 cell adhesion molecule (L1CAM) and hormonal receptors. Loss of asparaginase-like protein 1 (ASRGL1) has been shown to correlate with poor outcome in endometrial cancer. Our objective was to assess prognostication of endometrial cancer by ASRGL1 in conjunction with other available methodologies. Study DesignThis was a retrospective study of patients who underwent primary treatment at a single tertiary center. Tumors were molecularly classified by the Proactive Molecular Risk Classifier for Endometrial Cancer. Expression of ASRGL1, L1CAM, estrogen receptor, and progesterone receptor was determined by immunohistochemistry. ASRGL1 expression intensity was scored into four classes. ResultsIn a cohort of 775 patients, monitored for a median time of 81 months, ASRGL1 expression intensity was related to improved disease-specific survival in a dose-dependent manner (P < 0.001). Low expression levels were associated with stage II–IV disease and presence of uterine factors, i.e. high grade, lymphovascular space invasion, and deep myometrial invasion (P < 0.001 for all). Among the molecular subgroups, low expression was most prevalent in p53 abnormal carcinomas (P < 0.001). Low ASRGL1 was associated with positive L1CAM expression and negative estrogen and progesterone receptor expression (P < 0.001 for all). After adjustment for stage and uterine factors, strong ASRGL1 staining intensity was associated with a lower risk for cancer-related deaths (hazard ratio 0.56, 95 % confidence interval 0.32–0.97; P = 0.038). ASRGL1 was not associated with the outcome when adjusted for stage, molecular subgroups, L1CAM, and hormonal receptors. When analyzed separately within the different molecular subgroups, ASRGL1 showed an association with disease-specific survival specifically in “no specific molecular profile” subtype carcinomas (P < 0.001). However, this association became nonsignificant upon controlling for confounders. ConclusionsLow ASRGL1 expression intensity correlates with poor survival in endometrial cancer. ASRGL1 contributes to more accurate prognostication when controlled for stage and uterine factors. However, when adjusted for stage and other biomarkers, including molecular subgroups, ASRGL1 does not improve prognostic stratification.

Immunoreactivity of LMO7 and other molecular markers as potential prognostic factors in oropharyngeal squamous cell carcinoma

BackgroundDespite the better prognosis associated with human papillomavirus (HPV)-positive oropharyngeal squamous cell carcinoma (OPSCC), some patients experience relapse and succumb to the disease; thus, there is a need for biomarkers identifying these patients for intensified treatment. Leucine-rich repeats and immunoglobulin-like domain (LRIG) protein 1 is a negative regulator of receptor tyrosine kinase signaling and a positive prognostic factor in OPSCC. Studies indicate that LRIG1 interacts with the LIM domain 7 protein (LMO7), a stabilizer of adherence junctions. Its role in OPSCC has not been studied before.MethodsA total of 145 patients diagnosed with OPSCC were enrolled. Immunohistochemical LMO7 expression and staining intensity were evaluated in the tumors and correlated with known clinical and pathological prognostic factors, such as HPV status and LRIG1, CD44, Ki67, and p53 expression.ResultsOur results show that high LMO7 expression is associated with significantly longer overall survival (OS) (p = 0.044). LMO7 was a positive prognostic factor for OS in univariate analysis (HR 0.515, 95% CI: 0.267–0.994, p = 0.048) but not in multivariate analysis. The LMO7 expression correlated with LRIG1 expression (p = 0.048), consistent with previous findings. Interestingly, strong LRIG1 staining intensity was an independent negative prognostic factor in the HPV-driven group of tumors (HR 2.847, 95% Cl: 1.036–7.825, p = 0.043).ConclusionsWe show for the first time that high LMO7 expression is a positive prognostic factor in OPSCC, and we propose that LMO7 should be further explored as a biomarker. In contrast to previous reports, LRIG1 expression was shown to be an independent negative prognostic factor in HPV-driven OPSCC.

Abstract 65: Differential impacts of PD-L1 expression in tumor vs. stroma on survival in esophageal squamous cell carcinoma

Abstract Background: Esophageal squamous cell carcinoma (ESCC) has high incidence and recurrence rates, particularly in advanced stages. While the combined positive score (CPS) and tumor proportional score (TPS) are standard benchmarks for PD-L1 assessment in ESCCs, they face challenges such as reliance on subjective human interpretation and the uncertain clinical significance of strong staining intensities. Our study utilizes H-scores for PD-L1 expression and separates analysis of tumor and stroma, providing additional layers of information that complement current methods and contribute to a more comprehensive understanding of the role of PD-L1 in ESCC. Methods: This retrospective single-center cohort analysis involved 194 cases of ESCC who underwent surgical resection. We performed a comprehensive evaluation of PD-L1 expression within the tumor and stroma using whole slide images (WSI). QuPath image analysis software was used to classify regions as tumor or stroma and quantify the PD-L1 expression H-score. Results: Using optimal H-score cutoffs determined from the minimum P-value method, the Kaplan-Meier survival analysis showed that higher PD-L1 expression was significantly linked with better post-operative survival across all compartments. This correlation was most pronounced in the stroma (P&amp;lt;0.001). However, in multivariate Cox regression analysis, only high tumoral PD-L1 expression emerged as an independent predictor of favorable prognosis (HR=0.47, 95% CI: 0.27-0.81, P=0.007). The non-significant contribution of stromal PD-L1 in this model could be attributed to its association with advanced pT and pN stages, and the presence of lymphatic and venous invasion (P&amp;lt;0.001, respectively). Interestingly, stromal PD-L1 expression was associated with a longer duration of cytotoxic chemotherapy (cutoff 8 months, Median H-score 98.28 vs 55.24, P=0.026), but no significant correlation was observed with the duration of immune checkpoint inhibitor treatment. Conclusions: The study underscores the complex role of PD-L1 in ESCC, with tumoral PD-L1 expression emerging as an independent prognostic factor, and stromal PD-L1 expression found to be related with tumor progression and the duration of cytotoxic chemotherapy response. Citation Format: Tomohiro Murakami, Satoru Furuhashi, Yuuki Sakai, Kenichi Sekimori, Ryoma Haneda, Eisuke Booka, Tomohiro Matsumoto, Hirotoshi Kikuchi, Yoshihiro Hiramatsu, Hiroya Takeuchi. Differential impacts of PD-L1 expression in tumor vs. stroma on survival in esophageal squamous cell carcinoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 65.

Bcl-2 May Contribute to Evolution of Endometrial Hyperplasia, but It Isn’t a Factor in Subsequent Carcinogenesis

Endometrial carcinoma is common in the western world and is increasing in frequency in developing countries as well. Endometrial hyperplasia is considered to be a precancerous lesion. Apoptosis plays an important role in the neoplastic transformation of cells, with Bcl-2 being an anti-apoptotic cellular marker. Bcl-2 may have an important contribution to the development of endometrial carcinoma. The aim was to evaluate and compare the expression of markers Bcl-2 through the spectrum of normal endometrium, endometrial hyperplasias and endometrial adenocarcinomas. 50 cases were included in this study, comprising of 10 cases of normal endometrium, 10 cases of endometrial hyperplasia without atypia, 10 cases of atypical endometrial hyperplasia and 20 cases of endometrial adenocarcinomas from January 2017 to June 2018. Immunohistochemical staining with Bcl-2 was performed and the results were analyzed. Bcl-2 staining showed an increase in cases with strong intensity of staining from normal endometrium (20% cases) to cases of endometrial hyperplasia (75% cases). There was, however a decrease in the number of cases with strong intensity Bcl-2 staining as the lesions progressed from endometrial hyperplasia to endometrial carcinoma (30% cases). The results were statistically significant (P =0.00309). However, there was no significant association of staining between atypical hyperplasia and endometrial carcinomas (p=0.429), as well as the degree of carcinoma (p=0.6903). Bcl-2 expression showed an increase from cases of normal endometrium to endometrial hyperplasia supporting the process of increased anti- apoptotic activity in endometrial hyperplastic lesions. Its decreased expression in endometrioid adenocarcinoma when compared to endometrial hyperplasias could point towards other mechanisms of carcinogenesis other than failure of apoptosis.

Immunohistochemical evaluation of Glut1 in dentigerous cysts, odontogenic keratocysts, and ameloblastoma.

Glucose uptake may be considered the rate-limiting step for the growth and metabolism of the cancer cell. Studies on GLUT1 have shown that GLUT1 is involved in cell survival and proliferation in both healthy and pathological circumstances. GLUT1 expression is regarded as one of the crucial elements in the development of local aggressiveness, tumour invasiveness, and metastasis, particularly in malignant tumours. The role of glut1 in odontogenic cysts and tumours has remained uncertain. The aim of the study is to assess the expression of Glut1 in dentigerous cysts, odontogenic keratocysts, and ameloblastoma. The study was conducted in GSL Dental College. The study design was a resprospective immunohistochemical study. Formalin-fixed, paraffin-embedded blocks of histologically confirmed cases (n = 50), 10 cases of odontogenic keratocysts, dentigerous cysts, ameloblastomas solid, ameloblastomas unicystic, and dental follicles each. Brown colour staining was considered as positive staining for GLUT1. Quantitative analysis was performed by counting the number of labelled cells, and semi-quantitative analysis was conducted by assigning immunostaining intensity scores. Chi-square test was used to compare differences between the groups. A P value of ≤0.05 was considered as statistically significant. Odontogenic keratocysts and unicystic ameloblastoma showed ≥50% of label cells with strong intensity of staining. Odontogenic keratocysts and solid ameloblastoma showed sub-cellular localisation of staining in the cytoplasm and membrane. Dentigerous cysts exhibited combined nucleus, cytoplasm, and membrane sub-cellular localisation of staining. The development of ameloblastomas, odontogenic keratocysts, and dentigerous cysts appears to be influenced by GLUT-1. Variation in its expression may aid in explanation of some of the differences in biological activity of these lesions.

Investigating dysregulation of TGF-β1/SMAD3 signaling in atopic dermatitis: a molecular and immunohistochemical analysis.

Atopic dermatitis (AD) is a persistent and recurring inflammatory condition affecting the skin. An expanding corpus of evidence indicates the potential participation of transforming growth factor-β1 (TGF-β1) in the modulation of inflammation and tissue remodeling in AD. The primary objective of this study was to examine the aberrant modulation of TGF-β1/small mothers against decapentaplegic homolog 3 (SMAD3) signaling through a comprehensive analysis of their molecular and protein expression profiles. The study encompassed an aggregate of 37 participants, which included 25 AD patients and 12 controls. The assessment of mRNA and protein levels of TGF-β1 and SMAD3 was conducted utilizing quantitative real-time PCR and immunohistochemistry (IHC), whereas serum IgE and vitamin D levels were estimated by ELISA and chemiluminescence, respectively. Quantitative analysis demonstrated a 2.5-fold upregulation of TGF-β1 mRNA expression in the lesional AD skin (P < 0.0001). IHC also exhibited a comparable augmented pattern, characterized by moderate to strong staining intensities. In addition, TGF-β1 mRNA showed an association with vitamin D deficiency in serum (P < 0.02), and its protein expression was linked with the disease severity (P < 0.01) Furthermore, a significant decrease in the expression of the SMAD3 gene was observed in the affected skin (P = 0.0004). This finding was further confirmed by evaluating the protein expression and phosphorylation of SMAD3, both of which exhibited a decrease. These findings suggest that there is a dysregulation in the TGF-β1/SMAD3 signaling pathway in AD. Furthermore, the observed augmentation in mRNA and protein expression of TGF-β1, along with its correlation with the disease severity, holds considerable clinical significance and emphasizes its potential role in AD pathogenesis.

Abstract C022: Clinical, morphological, and immunohistochemical characterization of Nigerian prostatic adenocarcinomas: Current challenges and promising global initiatives

Abstract (A) AN INTRODUCTORY SENTENCE INDICATING THE PURPOSES OF THE STUDY: Prostate cancer, a major leading cancer among men worldwide, still poses a considerable medical and public health challenge in many parts of the world. Despite advances in screening and multimodal management of this disease, overall survival remains poor. The need to identify tumour markers as indicators for prognosis and targets for new therapeutic strategies, still remains a major challenge in prostate carcinoma research. The present study was conducted at a tertiary care teaching hospital at Uyo, South-South Nigeria to assess the expression of Ki-67 and p53 in histologically proven cases of prostatic carcinoma and to evaluate any correlation between the two as well as correlating the expression of both markers with tumour grading and scoring. (B) A BRIEF DESCRIPTION OF PERTINENT EXPERIMENTAL PROCEDURES: The study was conducted on120 histopathologically proven cases of prostate carcinoma received in ourTeaching Hospital, Uyo, South South Nigeria. Formalin fixed, paraffin embedded tissue blocks of prostate adenocarcinoma of those patients with complete clinical details were retrieved, stained and analysed for p53 and Ki67 proteins using immunohistochemistry. The results were analyzed. (C) A SUMMARY OF THE NEW, UNPUBLISHED DATA: The mean age of the prostate cancer patients was 61.7 years + 4.5 (range: 45–96 years). The most common pattern of Gleason grade seen was Group 5 (5+4) constituting 46.0% of the total cases while the Gleason score of 8-9 was the most predominant, constituting 65.0% of the total cases. The p53 positivity was observed in 31.0% of cases with percentage positive cells varying from 3-44% with mild, moderate and strong staining intensity. It was observed that with increase in the grade and score the number of cases showing positivity also increased with statistical significance seen with the same (p&amp;lt;0.05). The Ki-67 positivity was also observed in 10.0% of cases with percentage positive cells varying from 2-24% with moderate and strong staining intensity. It was observed that with increase in the grade and score the number of cases showing positivity also increased but no statistical significance was seen with the same. Five cases (3.3%) were positive for both p53 and Ki-67. Of the 5 cases of prostatic carcinoma positive for both Ki-67 and p53, 4 were seen in high grade tumours (80.0%) and there was statistical significance observed with increase in grade and score ((p&amp;lt;0.05).(D) CONCLUSIONS: The present study highlighted that the immunohistochemical expression of Ki-67 and p53 in prostate carcinoma was lower when compared to other studies. They also permit development of prognostic factors as their expression increases with increase in the grade and score as well as offering of appropriate targeted treatment leading to increase in the survival time. It is hoped that research in unraveling biology of prostatic adenocarcinomas among patients of African Ancestry (AA) and Caucasian Ancestry (CA) using more biomarkers will be carried out in the future. Citation Format: Emmanuel Kunle Abudu, Agility Ukyi-Osuwa Obi-Ihesie, Ikwo Jonathan Kudamnya, Oluwasayo Omolara Abudu. Clinical, morphological, and immunohistochemical characterization of Nigerian prostatic adenocarcinomas: Current challenges and promising global initiatives [abstract]. In: Proceedings of the 16th AACR Conference on the Science of Cancer Health Disparities in Racial/Ethnic Minorities and the Medically Underserved; 2023 Sep 29-Oct 2;Orlando, FL. Philadelphia (PA): AACR; Cancer Epidemiol Biomarkers Prev 2023;32(12 Suppl):Abstract nr C022.

Protein expression and localization of ABC transporters in pancreatic adenocarcinoma: Prognostic role of ABCC8

BackgroundATP-binding cassette (ABC) transporters translocate various substances across cellular membranes. Their deregulation may cause cancer drug resistance or perturbations in the supply of building blocks for cancer cells and modify patients’ prognosis. This study investigated protein expression and cellular localization of the previously suggested putative prognostic biomarkers – ABCB2/TAP1, ABCC7/CFTR, ABCC8/SUR1, and ABCD4 in patients with pancreatic ductal adenocarcinoma (PDAC). MethodsProtein expression and localization were assessed by immunohistochemistry in formalin-fixed paraffin-embedded primary tumor tissue blocks of 61 PDAC patients and associated with clinical data and the survival of patients. ResultsNo CFTR protein expression was observed in PDAC, while TAP1 and ABCC8 were expressed predominantly in the cytoplasm of tumor cells. Most samples (81 %) had detectable both membranous and cytoplasmic ABCD4 staining and 42 % had ABCD4 expressed in the apical orientation. Negative membranous ABCD4 staining was significantly more frequent in advanced stage III or IV tumors (p = 0.022). Small or medium counts of individual ABCC8-positive cells in the stroma surrounding tumor tubules were also more often found in stage III or IV (p = 0.044). Patients with moderate or strong ABCC8 cytoplasmic staining intensity in tumor cells had a 3.5-fold higher risk of disease progression than those with weak staining (p = 0.002). ConclusionsThe study shows for the first time that the cytoplasmic ABCC8 protein expression has prognostic value in PDAC.

FGFR3 and FGFR4 overexpression in juvenile nasopharyngeal angiofibroma: impact of smoking history and implications for personalized management.

Lifestyle factors, including smoking, have been linked to neoplastic diseases, and reports suggest an association between smoking and overexpression of FGFR (fibroblast growth factor receptor) in certain neoplasms. This study aims to assess the expression of FGFR3 and FGFR4 genes in patients with and without a history of smoking.A total of 118 participants were recruited, including 83 Juvenile Nasopharyngeal Angiofibroma (JNA) patients and 35 healthy participants, the JNA patients were further stratified as smokers and nonsmokers. Total RNA was extracted from the blood & saliva sample by using TRIzol reagent, and quantified using a Nanodrop, and then subjected to gene expression analysis of FGFR3/4 using RT-PCR. Immunohistochemistry analysis was employed using fresh biopsies of JNA to validate the findings. All experiments were performed in triplicates and analysed using the Chi-Square test (P < 0.05). Smokers exhibited significantly lower total RNA concentrations across all sample types (P < 0.001). The study revealed significant upregulation of both FGFR3/4 genes in JNA patients (P < 0.05). Moreover, FGFR3 expression was significantly higher among smokers 66% (95% CI: 53-79%) compared to non-smokers 22% (95% CI: 18-26%). Immunohistochemistry analysis demonstrated moderate to strong staining intensity for FGFR3 among smokers. The study highlights the overexpression of FGFR3/4 genes in JNA patients, with a stronger association observed among smokers. Furthermore, medical reports indicated higher rates of recurrence and bleeding intensity among smokers. These findings emphasize the potential role of FGFR3 as a key molecular factor in JNA, particularly in the context of smoking.

PRELIMINARY ANALYSIS OF STIM-1 EXPRESSION ON FORMALIN-FIXED PARAFFIN-EMBEDDED NASOPHARYNGEAL CANCER TISSUES

Nasopharyngeal cancer (NPC) is commonly diagnosed at the advanced stage with a poor prognosis. STIM-1 has become a promising cancer biomarker, especially in early diagnostic applications. There is a limited study on STIM1 expression in NPC, especially in formalin-fixed paraffin-embedded NPC tissues. Thus, the present work provides a preliminary analysis of STIM-1 expression on NPC tissues. This study aims to examine STIM-1 expression in NPC associated with tissue origin and the functional effects of STIM1 expression in NPC. Screening for target genes was using the KEGG PATHWAY database. The target gene analysis was initially done further studied using an immunohistostaining approach on NPC tissue samples. From the bioinformatic resources, STIM-1 has shown significant interaction with molecules network associated with cell migration and metastasis. Differentiated NPC showed moderate STIM-1 IHC staining intensity and undifferentiated NPC presented with strong STIM-1 IHC staining intensity. The present preliminary study suggests that STIM-1 could have a positive correlation with NPC pathogenesis. However, further comprehensive work using larger samples size is needed especially focusing on the STIM-1 expression and other clinicopathological parameters.

Phase 2 trial of zolbetuximab in combination with mFOLFOX6 and nivolumab in patients with advanced or metastatic claudin 18.2-positive, HER2-negative gastric or gastroesophageal junction adenocarcinomas.

TPS4173 Background: Zolbetuximab, a chimeric immunoglobulin G1 monoclonal antibody, binds to claudin 18.2 (CLDN18.2) and mediates tumor cell death through antibody-dependent cellular cytotoxicity and complement-dependent cytotoxicity. In the pivotal phase 3 SPOTLIGHT study, zolbetuximab with modified FOLFOX6 (mFOLFOX6; leucovorin [folinic acid], fluorouracil [5-FU], and oxaliplatin) significantly prolonged progression-free survival (PFS) and overall survival (OS) in patients with CLDN18.2-positive, HER2-negative gastric and gastroesophageal junction (G/GEJ) adenocarcinomas. The phase 2 ILUSTRO trial is investigating the efficacy and safety of zolbetuximab, alone and in multiple combinations, in patients with CLDN18.2-positive, HER2-negative advanced/metastatic G/GEJ adenocarcinomas. Methods: The ILUSTRO trial is enrolling two new cohorts, 4A (safety cohort) and 4B, with approximately 12 and 50 patients planned, respectively, to assess safety and efficacy of the first-line combination of zolbetuximab, mFOLFOX6, and nivolumab in G/GEJ adenocarcinomas. Both cohorts are enrolling patients whose tumors are HER2-negative with high or intermediate CLDN18.2 positivity (moderate to strong immunohistochemistry staining intensity in ≥75% of tumor cells [high] or ≥50% but &lt;75% of tumor cells [intermediate]). In Cohort 4A, patients will receive a loading dose of 800 mg/m2 of zolbetuximab with 240 mg of nivolumab and mFOLFOX6 on cycle 1 day 1, followed by 400 mg/m2 of zolbetuximab with 240 mg of nivolumab and mFOLFOX6 every 2 weeks (on days 15 and 29 of each 42-day cycle). Up to 12 mFOLFOX6 treatments (4 cycles) will be administered; patients may continue to receive folinic acid and 5-FU alongside zolbetuximab and nivolumab. Tolerability and safety of the combination of zolbetuximab, mFOLFOX6, and nivolumab will be evaluated during a 2-week dose-limiting toxicity assessment period. If the 800 mg/m2 loading dose of zolbetuximab was assessed as not tolerable, patients were to be de-escalated to 600 mg/m2 as their loading dose but received the same subsequent 400 mg/m2 doses of zolbetuximab every 2 weeks. Cohort 4B will receive this combination at the dose level determined in Cohort 4A. Efficacy endpoints include objective response rate, disease control rate, duration of response, PFS, and OS. Safety and tolerability, pharmacokinetics, immunogenicity, and health-related quality of life will also be evaluated. Currently, 20+ sites are recruiting in 6 countries (France, Italy, Japan, Korea, Taiwan, United States); more US sites are planned. Clinical trial information: NCT03505320 .

Evaluation of the Expression Intensity of Glucose Transporter-1 Marker and its Diagnostic Value in Differentiating Between Borderline and Malignant Ovarian Epithelial Tumors

Background: Ovarian cancer ranks second among gynecological cancers worldwide. This study aimed to compare the glucose transporter-1 (GLUT-1) expression in benign, borderline, and malignant ovarian epithelial tumors and evaluate GLUT-1 expression as a diagnostic tool for distinguishing tumors in the ovary. Materials and Methods: This descriptive-analytical cross-sectional study analyzed 69 pathological samples of patients diagnosed with ovarian epithelial tumors who underwent oophorectomy. Immunohistochemical staining was performed using GLUT-1 antibody. The intensity of cell membrane staining and the proportion of positive neoplastic cells were graded to score immunostaining. Chi-square/Fisher’s exact test was used to analyze the data. The sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV), and staining accuracy for GLUT-1 in distinguishing borderline from invasive tumors were calculated by standard methods (P&lt;0.05). Results: In all benign tumors, GLUT-1 staining was negative. In addition, weak staining intensity was observed in 38.5% of borderline tumors, and 96% of invasive tumors had strong staining intensity (P&lt;0.001). Strong GLUT-1 staining was found in 94.7% of Papillary Serous Carcinoma, 9/1% of Borderline Serous tumors, 100% of Brenner tumors, and clear cell carcinoma. The results demonstrated a high diagnostic value of GLUT-1 expression intensity in differentiating between borderline and malignant ovarian epithelial tumors (Accuracy: 97.10, Sensitivity: 96%, Specificity: 97.73). Conclusion: Overall, GLUT-1 expression could help distinguish benign from borderline, especially borderline from malignant ovarian epithelial tumors. Thus, it seems that it provides useful prognostic information, particularly for the borderline category.

Pathogenic role of Twist-1 protein in hydatidiform molar pregnancies and investigation of its potential diagnostic utility in complete moles

BackgroundComplete and partial moles (PM) are the most common gestational trophoblastic diseases. Due to some overlapping morphological findings, ancillary studies may be necessary.MethodsIn this cross-sectional study, 47 cases of complete mole (CM) and 40 cases of PM were randomly selected based on histopathological criteria. Only those cases that were agreed upon by two expert gynecological pathologists and confirmed by the P57 IHC study were included. The expression level of the Twist-1 marker in villi stromal cells, as well as syncytiotrophoblasts, was evaluated quantitatively (percentage of positive cells), qualitatively (staining intensity) and as a total comprehensive score.ResultsExpression of Twist-1 is higher and more intense in villous stromal cells of CMs (p < 0.001). Moderate to strong staining intensity in more than 50% of villous stromal cells, can differentiate CM and PM with 89.5% sensitivity and 75% specificity. In syncytiotrophoblasts of CM, Twist-1 expression was significantly lower than PM (p < 0.001). Negative or weak staining intensity in less than 10% of syncytiotrophoblasts, can distinguish CM and PM with 82.9% sensitivity and 60% specificity.ConclusionA higher expression of Twist-1 in villous stromal cells of hydatidiform moles is a sensitive and specific marker for the diagnosis of CMs. An elevated expression of this marker in villous stromal cells suggests another pathogenic mechanism for more aggressiveness of CMs in addition to the characteristics of trophoblast cells. The opposite result was obtained in the expression of Twist-1 in the syncytiotrophoblasts, compatible with defects in the process of formation of these supportive cells in CMs.

A study of RNA m6A demethylases in oral epithelial dysplasia and oral squamous cell carcinoma.

N6-Methyladenosine (m6A) modification is involved in the tumorigenesis of various cancers. However, the roles of RNA m6A demethylases, fat mass and obesity-associated protein (FTO), and AlkB homolog 5 (ALKBH5) in oral epithelial dysplasia (OED) and oral squamous cell carcinoma (OSCC) remain unclear. This study focuses on FTO and ALKBH5 expression by using immunohistochemistry. Twenty specimens each of OED, OSCC, and normal oral mucosa (NOM) were included. The expression pattern, the number of positive cells, the cell-staining intensity, and the histochemical scoring (H-score) were examined and analyzed. In all the OED and OSCC specimens, FTO and ALKBH5 were mainly expressed with moderate to strong staining intensity in the nuclei of the abnormal epithelial cells, respectively. Regarding the NOM, both RNA demethylases showed mild cell staining intensity and was present in 50-60% of the specimens. Interestingly, the percentage of cell positivity, the cell-staining intensity, and the H-score of the FTO and ALKBH5 in NOM, OED, and OSCC were increased, respectively (p<0.001). There was also a positive correlation between the FTO and ALKBH5 expressions in OSCC (r=0.62, p=0.003), but not in the NOM and OED. These results suggest a possible prognostic role of FTO and ALKBH5 expression in the malignant transformation of OED and tumor progression. Further studies are needed to elucidate the mechanisms underlying the roles of FTO and ALKBH5 in carcinogenesis.

Expression of SATB2 in primary cutaneous sarcomatoid neoplasms: a potential diagnostic pitfall

SATB2 can be used as an immunohistochemical marker for osteoblastic differentiation. The differential diagnosis of a cutaneous sarcomatoid neoplasm sometimes includes osteosarcoma when the tumour concomitantly involves the skin, soft tissue, and bone, or when there is a past medical history of osteosarcoma. As the utility of SATB2 immunohistochemistry in these scenarios was unclear, we aimed to determine the frequency and the pattern of SATB2 expression in a variety of cutaneous sarcomatoid neoplasms. SATB2 expression by immunohistochemistry was evaluated by intensity (0-3) and extent (0-100%) of staining to generate an h-score for each case. Expression levels were classified into high-positive (h-score ≥100), low-positive (20-99), and negative (<20) groups. Positive SATB2 expression was observed in 18/23 (78%) atypical fibroxanthomas (AFX), 10/19 (53%) pleomorphic dermal sarcomas, 9/20 (45%) cutaneous sarcomatoid squamous cell carcinomas, 14/39 (36%) sarcomatoid melanomas, 2/13 (15%) poorly differentiated cutaneous angiosarcomas, 10/17 (59%) high-grade cutaneous leiomyosarcomas, and 7/8 (88%) osteosarcoma controls. With the exception of AFX, all cutaneous neoplasms showed significantly lower average h-scores than osteosarcoma. AFX gave the highest average h-score (71) and percentage of high-positive cases (48%) among all examined cutaneous neoplasms. Only two (1.5%) of all cutaneous cases showed strong intensity of staining. Common SATB2 expression in various cutaneous sarcomatoid neoplasms poses a potential diagnostic pitfall when the differential diagnosis includes osteosarcoma. Requirement of strong staining and a high-positive h-score improves the specificity of SATB2 in differentiating these tumours from osteosarcoma.