Tropomyosin receptor kinase protein expression in Thai cholangiocarcinoma: Clinicopathological correlation, expression pattern, and prognosis.

OBJECTIVE: Tropomyosin receptor kinase (TRK) gene fusion was found in association with many tumors and could be a target of treatment. Immunohistochemistry (IHC) expression of TRK is widely used to screening this alternation. AIM: To investigate the expression of TRK protein detected by IHC in Thai cholangiocarcinoma (CCA) whereas the high endemic area of liver fluke infection and correlate with clinicopathological and survival data. METHODS: Retrospective study of CCA patients who diagnosed from January 2011 to December 2015. The TRK IHC was performed on paraffin-embedded tissues. RESULTS: A total of 85 CCA patients were enrolled. The mean age of the patients was 59 y (range; 35-79). Tumors were situated at intrahepatic (42 cases, 49.4 %), perihilar (41 cases, 48.2 %) and extrahepatic (2 cases, 2.4 %). The TRK IHC was expressed in 26 cases (31 %) and most of them (25 cases, 96.2 %) showed focal cytoplasmic expression with weak intensity. TRK IHC expression was not correlated with clinicopathological findings. Nevertheless, the median survival time of the TRK IHC positive and negative groups were 1.88-year and 1.30-year, respectively (p = 0.041) with the hazard ratio of 0.564 (p = 0.039, 95%CI 0.328-0.971). CONCLUSION: In Thai CCA, TRK IHC was detected about 1/3 of the patients and most expressed focally in the cytoplasm with weak staining. TRK expression showed better overall survival and was an independent prognostic factor. As the screening assays, the TRK IHC is wildly available with rapid, and high sensitivity but the confirmatory testing is necessary in tumors with low incidence of NTRK gene fusion.

Diagnosis of Metastatic Carcinoma Using Body Cavity Fluid Specimens: A Comparison of Diagnostic Panels.

INTRODUCTION: Body cavity effusions are routinely used as cytologic specimens. The distinction between metastatic carcinoma, mesothelioma, and reactive mesothelial cells remains a major challenge. Immunohistochemistry (IHC) is a supplemental method that can aid in diagnosis and often involves many markers as part of an IHC panel. Several immunohistochemical markers are now widely used. This study aims to determine the optimal immunomarkers and IHC panels to differentiate reactive mesothelial cells from metastatic cancer in body cavity fluid samples. METHODS: IHC was performed for claudin-4, MOC-31, Ber-Ep4, D2-40, and calretinin on sections derived from 152 cellblocks containing effusions. The samples consisted of 16 (10.53%) benign and 136 (89.47%) malignant tumors, including 87 (63.97%) lung cancers, nine (6.62%) breast cancers, 11 (8.09%) gynecologic cancers, seven (5.15%) pancreaticobiliary cancers, and 22 (16.17%) unspecified primary malignancies. RESULTS: Claudin-4, MOC-31, Ber-EP4, D2-40, and calretinin demonstrated sensitivities of 91.18%, 91.91%, 55.88%, 90.44%, and 98.53%, respectively. The corresponding specificities were 100.00%, 100.00%, 100.00%, 93.75%, and 100.00%. The sensitivity and specificity were both 100% when claudin-4 or MOC-31 was combined with calretinin. The combination of four markers as an IHC panel (claudin-4, MOC-31, calretinin, and D2-40) had a sensitivity of 97.79% and a specificity of 100.00%. CONCLUSION: Claudin-4 and MOC-31 both demonstrated significant diagnostic value in distinguishing metastatic epithelial carcinoma from reactive mesothelium. The sensitivity, specificity, and accuracy of these two markers, one of which is an epithelial marker and one of which is a mesothelial marker, reached 100%. Therefore, a combination of these two markers may be appropriate.

RET rearrangements are relevant to histopathologic subtypes and clinicopathological features in Thai papillary thyroid carcinoma patients.

Background: Papillary thyroid carcinoma (PTC) is the most common type of thyroid cancer. The RET gene rearrangements CCDC6::RET and NCOA4::RET are the most common RET gene rearrangements in PTC patients. Different RET::PTC rearrangements are associated with different PTC phenotypes. Methods: Eighty-three formalin-fixed paraffin-embedded (FFPE) PTC samples were examined. The prevalence and expression levels of CCDC6::RET and NCOA4::RET were determined using semi-quantitative polymerase chain reaction (qRT-PCR). The association of these rearrangements with clinicopathological data was investigated. Results: The presence of CCDC6::RET rearrangement was significantly associated with the classic subtype and absence of angio/lymphatic invasion (p < 0.05). While NCOA4::RET was associated with the tall-cell subtype, and presence of angio/lymphatic invasion and lymph node metastasis (p < 0.05). Multivariate analysis demonstrated that an absence of extrathyroidal extension and extranodal extension were independent predictive factors for CCDC6::RET, whereas the tall-cell subtype, large tumor size, angioinvasion, lymphatic invasion and perineural invasion were independent predictive factors for NCOA4::RET (p < 0.05). However, the mRNA expression level of CCDC6::RET and of NCOA4::RET were not significantly associated with clinicopathological data. Conclusion: CCDC6::RET was correlated with an innocent PTC subtype and characteristics, but NCOA4::RET correlated with an aggressive phenotype of PTC. Therefore, these RET rearrangements strongly associated with clinicopathological phenotypes and can be used as predictive markers in PTC patients.

CD44v6 expression in gastroenteropancreatic neuroendocrine neoplasms: Clinicopathological correlation and prognosis.

BACKGROUND: Clinical course of gastroenteropancreatic neuroendocrine neoplasm (GEP-NEN) patients are determined by complicated and multifaceted factors. Ki-67 has been used to predict the behavior of NEN with statistically significant high probability. Immunohistochemical prognostic markers other than Ki-67 have been reported, but due to insufficient validation, they have not been used routinely. CD44 and its isoform variants showed significant inverse correlation with lymph node status, distant metastasis, and mortality in pancreatic neuroendocrine neoplasm (Pan-NEN). The aim of this study is to determine the prognostic value of CD44v6 expression among GEP-NENs. MATERIALS AND METHODS: Immunohistochemical staining (IHC) of CD44v6 was studied in 26 formalin-fixed-paraffin-embedded (FFPE) specimens from GEP-NENs patients in the Srinagarind Hospital, Khon Kaen University, Thailand, between 2013 and 2018. Demographic data were collected. IHC staining result was interpreted based on H-score and correlated with neuroendocrine IHC markers, proliferation index, lymph nodes status, metastasis, and patients' survival. RESULTS: All cases showed variable staining for CD44v6 without statistically significant correlation with age, gender, histological grading, lymph node metastasis, distant metastasis, Ki-67 proliferative index, or neuroendocrine marker expression. Survival analysis showed association of higher CD44v6 expression with better prognosis (p = 0.033). Hazard ratio of CD44v6 low expression patients with age ≥ 49 was 12.736 (p = 0.028, 1.318-123.073), presence of lymph node metastasis was 8.267 (p = 0.036, 1.142-59.841), presence of distant metastasis 12.736 (p = 0.028, 1.318-123.073). CONCLUSION: Higher expression of CD44v6 IHC is significantly associated with better overall survival, histological grading, and Ki-67 index of GEP-NENs patients. CD44v6 IHC can be used as an additional prognostic immunohistochemical marker related to the metastatic status.

Chromosomal aberrations, visualized using UroVysion® fluorescence in-situ hybridization assay, can predict poor prognosis in formalin-fixed paraffin-embedded tissues of cholangiocarcinoma patients.

Cholangiocarcinoma (CCA) is a highly aggressive malignant tumor that has highest incidence in northeastern Thailand. The survival rate of CCA patients after receiving surgical treatment is quite low. Recently, genetic alterations including chromosome abnormalities have been studied as predictive factors and to aid planning for further treatment. This study aims to investigate the association between chromosomal aberrations, clinical data, and overall survival time of CCA patients. Formalin-fixed paraffin-embedded (FFPE) tissues from 194 CCA patients were examined. The copy numbers of chromosomes 3, 7, 17 and 9p21 were investigated using the UroVysion® fluorescence in-situ hybridization (FISH) assay. The overall survival time (OS) of CCA patients with or without polysomy of chromosomes 3, 7, 17 and/or loss of 9p21 were statistically analyzed in association with their clinicopathological parameters. Kaplan-Meier analysis was performed. The OS of patients with polysomy of chromosomes 3 + 7 was significantly shorter than those without this polysomy (log-rank P = 0.006; median OS 14.79 vs. 19.62 months). Moreover, patients with polysomy of chromosomes 3 + 7+17 and heterozygous for 9p21 loss have significantly shorter survival time than those without such chromosomal aberrations (log-rank P = 0.001; median OS 15.74 vs. 37.57 months). Interestingly, multivariate analysis revealed that polysomy of chromosomes 3 + 7 and of chromosomes 3 + 7+17 with 9p21 heterozygous loss were independent predictive factors of a poor OS (P = 0.027; P = 0.008, respectively).The chromosomal aberrations patterns which we evaluated using FISH; 1) polysomy of chromosomes 3 + 7 and 2) polysomy of chromosomes 3 + 7+17 with 9p21 heterozygous loss, have strong potential as indicators of poor prognosis in CCA patients.

Fluorescence in situ hybridization detection of chromosome 7 and/or 17 polysomy as a prognostic marker for cholangiocarcinoma.

Cholangiocarcinoma (CCA) is highly endemic in the Northeast Thailand. Recently, chromosome aberrations provided new insights into pathogenesis of CCA. Therefore, chromosome aberration might be used as a prognostic factor and therapeutic planning of this cancer. This aim of this study is to examine the correlation between an increase of chromosome 7 (C7) and/or 17 (C17) copy number variants (CNVs) with clinicopathological data and the overall survival time (OS) of CCA patients using fluorescence in situ hybridization (FISH) assays. C7 and C17 CNVs were examined using FISH form 157 formalin-fixed paraffin-embedded (FFPE) tissues of CCA patients from Khon Kaen, Thailand between 2011 and 2015. OS was visualized using Kaplan-Meier plot. Univariate and multivariate analyses were used to determine the ability of the clinicopathological parameters to predict OS. C17 > trisomy (odd ratio, 6.944, P < 0.001), C7/17 trisomy (odd ratio; 4.488, P = 0.019), and C7/17 > trisomy (odd ratio; 6.723, P < 0.001) were independently predictive factors for lymph node metastasis. Interestingly, an increase of C7, C17, and C7/17 CNVs in both trisomy and > trisomy was independently correlated with short median OS. An increased of C7 and/or 17 have a potential as a poor prognostic marker in CCA patients.