Applied Immunohistochemistry & Molecular Morphology最新文献

MammaPrint (MP) is a multigene assay utilized for prognostic and predictive use in early stage ER+/HER2-negative breast cancer (BC). There is limited data on MP correlation to histopathologic variables, including multivariable model Magee Equations (MEs). We compared pathologic variables to MP on 365 ER+/HER2-negative BCs in the adjuvant setting. Further, we analyzed MP and ME results in 26 core biopsy-proven ER+/HER2-negative BCs subjected to neoadjuvant chemotherapy (NACT). Post-NACT response was assessed by residual cancer burden (RCB) score. In the adjuvant cohort (n=365), high-risk (HR) MP correlated with high Nottingham score, low progesterone receptor H-score, ductal morphology, high Ki-67 index, and high ME score. In the neoadjuvant cohort (n=26), 24 were MP-HR and 2 were MP low-risk (MP-LR). The 2 MP-LR cases correlated with average ME scores of ≤25 and showed RCB scores of 2 or 3. Of the 24 MP-HR, 2 showed RCB-0 or RCB-1. Average ME>25 was seen in both of these cases and 7 others corresponding to an RCB-0 and RCB-1 rate of 22% (2 of 9) with ME>25 compared with 8% (2 of 24) with MP-HR. Lack of significant benefit from NACT (RCB-2 or RCB-3) was accurately predicted by average ME≤25 in 17 of 26 cases (65%) compared with only 2 of 26 cases (8%) with MP-LR. MP correlates with aggressive histopathologic features. While both MP and ME identified cases with robust chemotherapeutic response, MP overpredicted cases that would benefit. Therefore, MEs may more accurately predict response to NACT.

Transgelin-2 (TAGLN2) is an actin-binding protein associated with tumor progression, particularly in gastric and brain tumors. However, its role in ovarian cancer metastasis is still not fully understood. This study investigated the role of TAGLN2 and its potential mechanism in ovarian cancer metastasis. TAGLN2 expression in ovarian cancer and normal tissues was assessed using Oncomine, Human Protein Atlas (HPA), and immunocytochemistry (IHC). Skov3 cells with TAGLN2 knockdown (transient knockdown of TAGLN2 of homo TAGLN2 was performed using specific small interfering RNAs), and Skov3 cells transfected with scrambled siRNAs (negative control group) were subjected to colony formation and wound healing assays to assess cell proliferation and migration in vitro. RT-PCR and western blot were used to assess TAGLN2 mRNA and protein expression; immunofluorescence staining was used to investigate the TAGLN2 impact on the cytoskeletal organization. Elevated TAGLN2 levels were observed in ovarian cancer compared with normal tissues, which was confirmed by IHC of a tissue microarray containing 65 ovarian tumor samples. TAGLN2 knockdown reduced cell proliferation and migration in vitro. Also, TAGLN2 was found to co-localize with F-actin; the knockdown of TAGLN2 impaired cytoskeletal organization, emphasizing its influence on cellular structures. In summary, TAGLN2, highly expressed in ovarian cancer, promotes migration and proliferation through cytoskeletal reorganization; thus, it may be a new therapeutic target for ovarian cancer.

Human telomerase reverse transcriptase (hTERT) is detectable in normal progenitor cells, tumor cells, and B-cell chronic lymphocytic leukemia (B-CLL) cells. hTERT expression, in addition to other prognostic factors, is reportedly associated with a poor prognosis in B-CLL. In this study, we aimed to analyze and compare hTERT immunoexpression in B-CLL bone marrow (BM) lymphocytes and benign pleural effusion lymphocytes. Standard cytologic analysis and immunocytochemical assessment of hTERT immunoexpression were performed in BM lymphocytes from 25 patients with B-CLL and pleural effusion lymphocytes from 18 patients with pneumonia and effusion-reactive lymphocytosis. The percentages and score values of hTERT nucleus (TN)-immunopositive BM lymphocytes in patients with CLL were significantly higher than those for reactive effusion lymphocytes with no or few TN-immunopositive lymphocytes. The appearance of TN immunopositivity in CLL lymphocytes showed mainly numerous prominent or large dots, and diffused TN immunopositivity was detected; in contrast, TN-immunopositive benign effusion lymphocytes had one or few immunopositive nuclear dots. Further investigations are needed to clarify whether lymphocyte TN immunopositivity can reveal subgroups of patients with CLL with a worse prognosis and whether there is a reliable difference in TN immunopositivity between CLL and benign effusion lymphocytes.

Thymomas are rare mediastinal tumors exhibiting heterogeneous behavior. Although histologic subtypes and stages serve as prognostic factors, the molecular mechanisms of thymoma progression are unclear. Immunohistochemical markers like Bcl-2, D2-40, β-catenin, and E-cadherin offer insights into thymoma biology, but their predictive value for clinical outcomes remains uncertain. This study evaluated the expression of these markers across thymoma subtypes and stages, aiming to assess their prognostic significance. A retrospective analysis was conducted on 66 thymoma cases resected at a single center between 2005 and 2023. Immunohistochemical staining was performed to assess the expression of Bcl-2, D2-40, β-catenin, and E-cadherin. Clinicopathological characteristics were correlated with immunohistochemical findings using statistical analysis. Differential expression patterns of Bcl-2, D2-40, β-catenin, and E-cadherin were observed across thymoma subtypes and clinical stages. Bcl-2 displayed cytoplasmic positivity predominantly in type A and B thymomas, while E-cadherin showed membranous staining in type B thymomas and cytoplasmic staining in type A and AB thymomas. β-catenin demonstrated membranous staining in type B thymomas and cytoplasmic staining in type A and AB thymomas. D2-40 expression was localized to peripheral regions and invasive nests of thymomas, with higher expression in type B2 thymomas and early-stage tumors. Our findings indicate that immunohistochemical markers may provide valuable insights into thymoma biology and prognosis. Further validation in larger, multicenter cohorts is warranted to confirm the prognostic significance of these markers and their potential utility in guiding clinical management.

High-grade serous carcinoma is categorized based on p53 mutation status. A relationship is known to exist between p53 mutations and p53 immunoexpression patterns, including overexpression, complete absence, cytoplasmic, and wild-type patterns. The ubiquitin ligases WWP1 and PARC, known to regulate p53 activation, are hypothesized to influence the pathogenesis of serous ovarian tumors. This retrospective study examined 7 low-grade serous carcinomas, 38 high-grade serous carcinomas, and 15 serous cystadenomas, with immunohistochemical analyses performed for WWP1, PARC, and p53. High-grade serous carcinoma cases were classified into wild-type, cytoplasmic, complete absence, or overexpression categories based on p53 immunohistochemistry. PARC and WWP1 expressions were compared across p53 categories and diagnoses. Results showed a statistically significant reduction in WWP1 and PARC expression in serous carcinomas, with the most pronounced loss observed in high-grade cases. Among morphologically classified high-grade carcinomas, 17 overexpression, 11 complete absence, 6 wild-type, and 4 cytoplasmic p53 cases were identified. A statistically significant relationship was found between PARC, WWP1, and p53 status. Higher expression levels of PARC and WWP1 were detected in p53 wild-type cases, whereas lower expression levels were associated with cases exhibiting p53 overexpression and complete absence. This study suggests that PARC and WWP1 play a role in the pathogenesis of high-grade serous ovarian carcinoma, potentially mediated by p53, making them promising targets for treatment and prognostic markers in serous ovarian cancer.

Coilin is the signature protein of Cajal bodies (CBs), membrane-less organelles probably acting as sites for post-transcriptional RNA modification. Recent data suggest that coilin may be a regulator of the NF-kB activity, and Hodgkin lymphomas are hallmarks of neoplasms with NF-kB dysregulation. To the best of our knowledge, the immunohistochemical expression of coilin has been never investigated in Hodgkin lymphomas. We herein examined, by immunohistochemistry, full tissue sections of 58 classical Hodgkin lymphomas diagnosed in 31 male and 27 female patients and found that none of the cases expressed coilin. We compared these findings with Coilin expression in diffuse large B-cell lymphomas (DLBCL), where the marker was also negative. This finding represents the first data on coilin in lymphomas and prompts further studies to explore this downregulation.

A major goal in Alzheimer disease (AD) research is the reduction of the abnormal tau burden. Using multispectral analyses on brain tissues from humans who died of AD it was documented that neurons with hyperphosphorylated tau protein accumulate many proteins of the BCL2 family, including those that block cell turnover (eg, BCL2, MCL1, BCLXL) and those that promote cell turnover (eg, NOXA, PUMA, BAK, BAX). A mouse model of AD with the humanized hyperphosphorylated tau protein was used to test the hypothesis that shifting this balance to a pro-cell turnover milieu would reduce the tau burden with concomitant clinical improvement. Here, we show that a mouse model of AD with death at 11 to 15 months due to CNS tauopathy had a marked reduction in the tau burden after treatment with the FDA-approved drug venetoclax, which blocks BCL2. The reduction of the number of target neurons positive for hyperphosphorylated tau protein after venetoclax treatment in the brain and spinal cord neurons was 94.5% as determined by immunohistochemistry and 98.1% as documented with the modified Bielchowsky stain. The venetoclax treatment began after documented neurofibrillary tangles (NFTs) were evident and there was a concomitant reduction in neuroinflammation. The treated mice were robust until sacrificed at 13 months as compared with the untreated mice that showed unequivocal evidence of brain and spinal cord damage both clinically and at autopsy. We conclude that otherwise inexorable abnormal tau protein deposition, even after initiation, can be prevented by a drug that blocks one anti-cell turnover protein abundant in the NFTs of human AD.

Despite improvements in machine learning algorithms applied to digital pathology, only moderate accuracy, to predict molecular information from histology alone, has been achieved so far. One of the obstacles is the lack of large data sets to properly train machine learning models. We therefore built a data set of 185,538 breast cancer (BC) including hematoxylin and eosin (H&E) and associated immunohistochemistry (IHC) images of the proliferative marker Ki67, estrogen receptor (ER), progesterone receptor (PR), and the human epidermal growth factor receptor 2 (HER2). Optimal registration of H&E and IHC pairs was achieved. Ki67, ER, and PR IHC labels, to be predicted, were extracted from IHC assays using image analysis. These labels were ordinaly classified with incremental thresholds (cumulative logit models with balanced and partial proportional odds). HER2 label was determined as follows: positive if tumor IHC 3+ pattern is identified and otherwise negative. Cases with IHC equivocal score (2+) were excluded. A vision transformer (ViT)-based pipeline, trained with this data set, achieved prediction performance of 90% in terms of area under the curve (AUC) of the receiver operating characteristic (ROC) curves. ViT outperformed the weakly supervised clustering-constrained attention multiple instance learning (CLAM) which was developed to automatically identify subregions of high diagnostic value in whole slide. As a first step to "explain" artificial intelligence (AI), we evaluated the ability of both classifiers to localize these high diagnostic value subregions by inspecting their respective "attention" heat-maps. Despite high ViT AUC-ROC results, heat-maps do not obviously match areas of high diagnostic value subregions; it might however provide direction for future work to improve AI attention within whole slide images. Our proposed data set is publicly available.

We aimed to investigate molecular mechanisms affecting melanoma progression by comparing genetic/epigenetic features between melanomas of different Breslow thickness and stage using TCGA (The Cancer Genome Atlas) data. The TCGA, Firehose Legacy, melanoma data set was utilized on the cBioPortal website. The cases were compared in terms of mRNA expression and DNA methylation. Gene Ontology (GO) and KEGG pathways enrichment analysis were performed using the online WebGestalt tool. STRING and Cytoscape software were used to construct a protein-protein interaction network and identify hub genes. P and q <0.05, FDR< 0.05 were considered statistically significant. 1001 differentially expressed genes were identified between thin (≤1 mm) and thick (>1 mm) melanomas. Pathway analyses revealed that genes enriched in thin melanomas were associated with adaptive immune response, T-cell activation, immune response regulation, leukocyte, and cytokine-related pathways, whereas genes enriched in thick melanomas were related to epidermis development. Ten hub genes were identified ( CD4, IFNG, PTPRC, CD8A, CTLA4, CD69, ICOS, CD27, CD28, CD19 ). All of these genes are involved in crucial immunological processes. Understanding the complex changes in melanoma progression is essential for accurate diagnosis and prediction of prognosis. Our results may shed light on subsequent studies to identify the steps in melanoma progression.