Modern Pathology最新文献

Molecular studies have shown alternative lengthening to telomeres (ALT) to be an important prognostic biomarker of shorter relapse-free survival (RFS) for patients with pancreatic neuroendocrine tumors (PanNETs) and other neoplasms. However, the preferred method of detecting ALT in tissue is by fluorescence in situ hybridization (FISH), which has several clinical limitations. These issues necessitate the creation of a chromogenic ALT assay that can be easily implemented into routine practice. A chromogenic in situ hybridization (CISH) assay was developed using genetically modified osteosarcoma cell lines, 20 normal pancreata, 20 ALT-positive PanNETs, and 20 ALT-negative PanNETs. Thereafter, it was validated on a multiinstitutional cohort of 360 surgically resected PanNETs and correlated with multiple clinicopathologic features, RFS, and FISH results. Separately, 109 leiomyosarcomas (LMS) were evaluated by both CISH and FISH, and, similarly, the prognostic significance of ALT status was assessed. Upon optimization, ALT-CISH was identified in 112 of 360 (31%) primary PanNETs and was 100% concordant with FISH testing. ALT correlated with several adverse prognostic findings and distant metastasis (all P < .004). The 5-year RFS for patients with ALT-positive PanNETs was 35% as compared with 94% for ALT-negative PanNETs. By multivariate analysis, ALT was an independent prognostic factor for shorter RFS. Similarly, ALT was associated with shorter RFS in patients with LMS and, analogous to PanNETs, a negative, independent prognostic factor. ALT-CISH was developed and validated in not only PanNETs but also sarcomas, specifically LMS. CISH testing has multiple advantages over FISH that facilitate its widespread clinical use in the detection of ALT and prognostication of patients with diverse neoplasms.

Human epidermal growth factor receptor-2 (HER2) expression is an important biomarker for the management of RAS wild-type metastatic colorectal carcinoma (CRC). Immunohistochemistry (IHC) with reflex in situ hybridization (ISH) is accepted as a standard method of assessment, yet there are currently the following 2 sets of criteria used to interpret results: the HER2 Amplification for Colorectal Cancer Enhanced Stratification (HERACLES) criteria and the MyPathway criteria. The HER2 Amplification for Colorectal Cancer Enhanced Stratification criteria require ISH confirmation when IHC staining is 3+ in 10% to 49% of cells, whereas the MyPathway criteria mirror those for gastric HER2 assessment and do not recommend ISH confirmation in the previously referenced scenario. We aimed to assess the prevalence of HER2 3+ heterogeneity and its association with ERBB2 copy number amplification to evaluate the necessity of ISH testing when IHC staining is 3+ in <50% of cells. Next-generation sequencing of DNA (592-gene panel or whole exome sequencing) was performed for 13,208 CRC tumors submitted to Caris Life Sciences. HER2 (4B5) expression was tested using IHC. A subset of tumors was tested for ERBB2 amplification via chromogenic ISH and/or via next-generation sequencing (copy number amplification). χ² tests or Fisher exact tests were applied where appropriate, with P values adjusted for multiple comparisons (P < .05). Of 13,208 CRCs with HER2 IHC, 87.4% (11,541/13,208) were negative for HER2 expression (≤3+ intensity and <10% tumor-cell staining) and 11.2% (1473/13,208) demonstrated at least low HER2 expression (1 to 2+ and ≥10%). Only 1.5% (194/13,208) of all tested tumors were either positive or heterogeneously positive for HER2 overexpression (3+ and ≥10%). Of these, 14% (28/194) had heterogenous HER2 overexpression (3+ staining of 10%-49% of cells). Among 22 HER2-positive/heterogenous cases with successful ISH testing, 100% (22/22) demonstrated amplification via ISH. Because the classification of tumors as HER2-positive/heterogenous using IHC correlated very closely with ISH positivity, our results suggest that ISH is likely unnecessary for CRCs with 3+ HER2 overexpression in 10% to 49% of neoplastic cells.

The immune landscape of hormone receptor-positive, human epidermal growth factor receptor 2-negative metastatic breast cancer (HR+/HER2- mBC), the most common subtype of BC, remains understudied. This is mainly because of reduced sample acquisition opportunities from metastases as compared with primary tumors. In this study, we explored stromal tumor-infiltrating lymphocytes (sTIL) in metastatic samples collected through our post-mortem tissue donation program UZ/KU Leuven Post-mortem Tissue Donation program to Enhance Research (NCT04531696). sTIL were scored as a continuous parameter according to the international guidelines on 427 metastases and 38 primary untreated tumors acquired from 20 patients with HR+/HER2- mBC. Estrogen receptor (ER) status was evaluated on 362 metastases with a cutoff value for positivity set at 1% according to the American Society of Clinical Oncology/College of American Pathologists guidelines. Our analyses show that 54% and 15% of metastases had sTIL levels of ≥1% and ≥5%, respectively. sTIL levels tended to be lower in metastases as compared with their respective primary tumors (estimate, -2.83; 95% CI, -5.77 to 0.11; P = .07). sTIL levels were lower in metastases from invasive lobular carcinoma than in metastases from invasive breast carcinoma of no special type (estimate, -1.67; 95% CI, -2.35 to -0.98; P < .001). A loss of ER expression was observed in 14% of all metastases, yet a negative ER status was not significantly associated with increased sTIL levels. Finally, sTIL levels were significantly higher in lung and axillary lymph node metastases compared with all metastases. Although these analyses were conducted on multiple metastases obtained at the end of life after several lines of treatment, the data provide novel and valuable insights into the state of immune infiltration in patients with HR+/HER2- mBC.

Accurate and rapid screening of adenocarcinoma cells in serous cavity effusion is vital in diagnosing the stage of metastatic tumors and providing prompt medical treatment. However, it is often difficult for pathologists to screen serous cavity effusion. Fixed agglutination cell block can help to improve diagnostic sensitivity in malignant tumor cells through analyzing larger volumes of serous cavity effusion, although it could accordingly lead to screening of more cells for pathologists. With the advent of whole slide imaging and development of artificial intelligence, advanced deep learning models are expected to assist pathologists in improving diagnostic efficiency and accuracy. In this study, so far as we know, it is the first time to use cell block technology combined with a proposed weakly supervised deep learning model with multiple instance learning method to screen serous adenocarcinoma. The comparative experiments were implemented through 5-fold cross-validation, and the results demonstrated that our proposed model not only achieves state-of-the-art performance under weak supervision while balancing the number of learnable parameters and computational costs and reduces the workload of pathologists but also presents a quantitative and interpretable cellular pathologic scene of serous adenocarcinoma with superior interpretability and strong generalization capability. The performances and features of the model indicate its effectiveness in the rapid screening and diagnosis of serous cavity effusion and its potential in broad clinical application prospects, eg, in precision medical applications. Moreover, the constructed 2 real-world pathologic data sets would be the first public whole slide imaging data sets of serous cavity effusion with adenocarcinoma based on cell block sections, which can help assist colleagues.