BackgroundThe spatial distribution and functional heterogeneity of tumor-infiltrating lymphocytes (TILs) significantly impact patient outcomes in non-small cell lung cancer (NSCLC). While T cell factor 1 (TCF1) expressing TILs have emerged as key players in sustaining anti-tumor immunity, their subset characterization, localization, and clinical significance within the tumor microenvironment remain poorly defined.MethodWe performed multiplex immunohistochemistry and immunofluorescence to characterize TCF1+ immune cell subsets, in 102 NSCLC tumors, separately analyzing the tumor center (TC) and invasive front (IF). We integrated this data with publicly available single-cell RNA-sequencing datasets and clinical outcome analyses.ResultsCD4+ T cells and CD79α+ B cells, dominate the TCF1+ landscape, while CD8+ T cells constitute a minority of TCF1+ immune cells, particularly in the TC. We demonstrated the presence of tumor-infiltrating IgG+/IgA+ plasma cells co-expressing TCF1. PD1+TCF1- cells were more frequent than PD1+TCF1+ cells both in the TC and IF, reflecting that terminally differentiated exhausted TILs predominate within the tumor microenvironment. Survival analyses revealed significantly different prognostic impact of TILs including TCF1-expressing cells based on topography. Multivariate analysis showed that increased CD8+TCF1+ cells (HR: 2.5; p=0.039) and increased TCF1 expression by cancer cells (HR: 2,7; p=0.041) in the TC and CD4+TCF1+ cells (HR: 0.4; p=0.043) in the IF emerged as negative and positive independent prognostic markers for Disease-free survival (DFS), respectively. Integrating PD-L1 expression with TILs, PD-L1 immunopositivity was correlated with increased CD8+ and PD1+TCF1- cell infiltration and was associated with favorable DFS especially in the TC.ConclusionsOur findings support a more refined framework for TCF1+ TIL assessment and TCF1 expression across cellular populations in the tumor microenvironment, with implications for prognostication in operable NSCLC.

BACKGROUND:Recently, a molecular characterization of adenoid cystic carcinoma (ACC) has been proposed, based on upregulation of c-Myc, a proto-oncogene involved in carcinogenesis, and p63, a transcription factor of the p53 gene family. The aim of this study was to evaluate c-Myc and p63 expression and classify patients with ACC into the proposed molecular groups. METHODS:We included in the analysis 50 patients with ACC of the head and neck region diagnosed treated between 2000 and 2021 in four tertiary referral centers in Greece. Patient demographics and disease characteristics were retrieved from patients' medical records. C-Myc and p63 expression were evaluated by immunohistochemistry. RESULTS:Forty-seven patients were eligible for classification. The majority of patients had primary non-metastatic tumors. P63 protein was found to be overexpressed in 39 patients (78 %), who were classified as ACC type II (ACC-II). Eight patients with no p63 overexpression were classified as ACC type I (ACC-I). Among ACC-II patients, 31 had negative expression of c-Myc and 8 patients had low c-Myc expression. We found a statistically significant difference in the primary tumor location between the two groups (p = 0.0470). Notably, patients that belonged to the ACC-II subgroup had more favorable prognosis (50 months for ACC-I vs. 135 months for ACC-II, p = 0.0066). CONCLUSIONS:We confirmed the recently published data regarding the c-myc/p63 prognostic classification for ACC. Evaluation of c-Myc and p63 should be routinely performed in ACC tumors to allow for accurate stratification and implementation of ACC subtyping for clinical trials.

Invasive lobular carcinoma (ILC) is the most common special type of breast carcinoma, accounting for 5–15% of all breast carcinoma cases. Its metastatic pattern differs from that of invasive breast carcinoma of no special type, with ILC metastases to the peritoneum, gastrointestinal tract, and female genital tract being more frequent. This literature review focuses on ILC cases with metastasis to the female genital tract (FGT). Searches were conducted in medical databases including PubMed, Scopus, and Web of Science, using specific keywords. Inclusion criteria centered on studies presenting one or more cases of patients with ILC metastasis to the FGT and English language publications. Exclusion criteria included articles that did not present original research findings, studies with insufficient data, and publications in languages other than English. A thorough analysis of 154 results from PubMed, 56 from Scopus, and 173 from Web of Science after the application of inclusion and exclusion criteria resulted in the inclusion of 54 manuscripts describing 61 cases. The demographic, clinicopathological, and therapeutic aspects of ILC metastases to the FGT were reviewed and the differential diagnosis and prognosis for each anatomic location in the FGT were discussed separately. Our analysis of the data showed that the restricted mean survival time was 186 ± 30.7 months and that a negative ER on a secondary tumor was found to be linked to worse patient survival rates. Also of note is the fact that in 37.7% of cases there was involvement of multiple FGT anatomic locations and in 36% of cases there were metastases in organs or anatomic locations other than the FGT. To our knowledge, our study is the only one to describe the features of patients with secondary FGT involvement from ILC.

DICER1 syndrome (DICERs) represents a tumor predisposition genetic syndrome, inherited in an autosomal dominant manner. Germline loss-of-function variants of the DICER1 gene lead to impaired processing of microRNA, gene expression, and increased risk of tumorigenesis. Although pleuropulmonary blastoma (PPB) is the hallmark of the syndrome, multiple extrapulmonary malignant and non-malignant conditions have also been described, including multinodular goiter (MNG) and sex-cord stromal tumors. MNG is one of the most common components and is associated with an increased risk of thyroid carcinoma. Sertoli-Leydig cell tumor (SLCT) represents the most prevalent type of sex-cord stromal tumor associated with the syndrome, whereas juvenile granulosa cell tumor (JGCT) is considered to be a very rare phenotype. They both may present with abdominal pain due to mass effect and menstrual irregularities in case of hormone production. Although they exhibit low rates of mortality, recurrence rates highly depend on the grade of malignancy. Herein, we report a novel pathogenic DICER1 variant associated with MNG, bilateral ovarian SLCT, and JGCT in a young Greek patient. Clinicians should be aware of a potential germline DICER1 variant when evaluating MNG in young patients, especially if it coexists with other neoplasms.

The World Health Organization (WHO) 2019 classification of hepatocellular carcinoma (HCC) introduces new histological subtypes aiding morphomolecular classification. We aimed to reclassify and restage archival HCC following central pathology review and evaluate the frequency of new subtypes in a large European HCC cohort. All histological material of 100 curative liver resection specimens for HCC from 100 patients (80 male, median age 72, IQR: 61.5–76.5 years, BCLC stage 0-C) operated from 2001 to 2018 was centrally reviewed. All cases were reclassified according to WHO 2019 and restaged according to TNM 2017 system. Thirty-five HCCs (35

Invasive lobular carcinoma (ILC) is the most common special type of breast carcinoma, accounting for 5-15% of all breast carcinoma cases. It exhibits several unique features in morphology, clinical behavior, radiological features, molecular characteristics, and metastatic pattern. The latter differs from that of invasive breast carcinoma of no special type with ILC metastases to the peritoneum, gastrointestinal tract, and female genital tract being more frequent. We herewith review the demographic, clinicopathological, and therapeutic aspects of ILC metastases to the female genital tract and discuss separately the differential diagnosis and prognosis for each anatomic location of the female genital tract.

Tumour antigen-specific CD8+CD39+ T cells (TSTs) are crucial components of anti-tumour adaptive immunity, which in tumour microenvironment (TME) may be found at various differentiation stages, from early stem-like states to terminal stages of exhaustion. Our aim was the evaluation of TSTs quantitative and qualitative characteristics in hepatocellular carcinoma (HCC) TME, depending on whether they exhibit stem-like or exhaustion features. We analyzed 100 HCC patients undergoing curative hepatectomy. Multiplex immunohistochemistry (CD8/CD39/TCF1, CD8/CD39/TOX) was applied on tissue microarray sections with digital imaging used to evaluate T cell infiltration in tumour center, periphery and adjacent non-neoplastic liver. Tertiary lymphoid structures (TLS) were assessed on H E-stained slides. In HCC TME, 72

Background/Objectives: There are limited data regarding immunohistochemical profiling of immune cells in bone marrow trephine biopsies of patients with high-risk myelodysplastic syndromes (HR-MDS). Methods: We sought to objectively quantify, with the use of digital pathology, the density (cells/mm2) of the prominent adaptive immunity cell populations in sixty-four (64) bone marrow trephine biopsies of HR-MDS patients receiving 5-Azacytidine. We focused on CD3(+) T cells, CD8(+) cytotoxic T cells (Tc), helper T cells (Th), Foxp3(+) regulatory T cells (Tregs), CD20(+) B-cells and CD138(+) plasma cells and evaluated the presence and the number of lymphoid aggregates. A control group of twenty “non-MDS” patients was included in the study. Results: We identified a significant decrease in adaptive immune cell densities in the HR-MDS patients compared to the non-MDS controls. Increased T and Th cell densities correlated with the response to 5-Azacytidine (5-AZA) treatment. Higher T, Tc, Th and plasma cells densities and low B, Tregs and Tregs/T cells ratios correlated with increased overall survival. Reduced Tregs, Tregs/T cells, Tregs/Tc and plasma cells showed improved leukemia-free survival. A modified IPSS-R (IPSS-R-I), combining the initial IPSS-R with the immune populations’ parameters, improved overall survival and showed a double-fold increase in Cox calculated hazard ratios. Conclusions: Immunohistochemical bone marrow immune profiling represents a powerful and easily useable tool for investigating the possible role of bone marrow immune microenvironment in the pathogenesis and progression of MDS, but also its association with the response to 5-AZA treatment and clinical outcomes.

In the setting of pronounced inflammation, changes in the epithelium may overlap with neoplasia, often rendering it impossible to establish a diagnosis with certainty in daily clinical practice. Here, we discuss the underlying molecular mechanisms driving tissue response during persistent inflammatory signaling along with the potential association with cancer in the gastrointestinal tract, pancreas, extrahepatic bile ducts, and liver. We highlight the histopathological challenges encountered in the diagnosis of chronic inflammation in routine practice and pinpoint tissue-based biomarkers that could complement morphology to differentiate reactive from dysplastic or cancerous lesions. We refer to the advantages and limitations of existing biomarkers employing immunohistochemistry and point to promising new markers, including the generation of novel antibodies targeting mutant proteins, miRNAs, and array assays. Advancements in experimental models, including mouse and 3D models, have improved our understanding of tissue response. The integration of digital pathology along with artificial intelligence may also complement routine visual inspections. Navigating through tissue responses in various chronic inflammatory contexts will help us develop novel and reliable biomarkers that will improve diagnostic decisions and ultimately patient treatment.

Lymphoepithelial or lymphoepithelioma-like carcinoma is a poorly differentiated carcinoma located outside the nasopharynx with similar morphologic characteristics to its nasopharyngeal counterpart. Lymphoepithelial carcinoma of the lung is a rare subtype of squamous cell lung carcinoma frequently associated with Epstein-Barr virus (EBV) infection, accounting for approximately 1% of non-small cell lung carcinomas (NSCLC). We herewith present a case of a 78-year-old female patient who was diagnosed with lymphoepithelial carcinoma of the lung, emphasizing its distinct epidemiological features, clinical workup, and histopathological characteristics. Furthermore, we discuss its histologic differential diagnosis. Finally, we refer to this tumor's unique molecular and immunological profile and its treatment modalities, and we review the literature.

Simple Summary Malignant Brenner tumors are rare ovarian neoplasms. Our aim is to provide insights concerning this rare entity. We reviewed 115 cases reported in the English literature until 15 September 2023, and analyzed the available demographic, clinical, and pathologic data. We also described the treatment modalities. A comparison of the available data showed that patients treated with lymph node dissection had a better disease-related survival rate. Disease recurrence was associated with tumor stage with marginal statistical significance and was more frequent in patients with ascites and those with abnormal CA-125 levels. Larger series with treatment details and long term follow-up data are needed to define the optimal management for this uncommon entity.Abstract Background: Malignant Brenner tumors are rare ovarian tumors, accounting for less than 1% of malignant ovarian neoplasms. The aim of this manuscript is to systematically review the current literature concerning malignant Brenner tumors. Methods: We searched three medical databases (PubMed, Scopus, and Web of Science) for relevant articles published until 15 September 2023. Results: After applying inclusion and exclusion criteria, 48 manuscripts describing 115 cases were included in this study from the English literature. Conclusions: We analyzed the demographic, clinical, pathological, and oncological characteristics of 115 patients with malignant Brenner tumors. The statistical analysis showed that recurrence was marginally statistically significantly related to tumor stage and was more common in patients with ascites and in women with abnormal CA-125 levels; patients that were treated with lymphadenectomy had better disease-specific survival.

Introduction: Endometriosis is a common benign condition affecting 10-15% of women of reproductive age. An unusual site of endometriosis is the canal of Nuck, which is a physiologically obliterated space in women spanning the area from the deep inguinal ring to the labia majora. Case presentation: A 37-year-old woman, with a past medical history of several in vitro fertilization attempts, presented with a right-sided painful inguinal mass. She was subsequently offered surgical exploration and excision of the lesion, which revealed the presence of endometrial glands and stroma. Discussion: Despite being a relatively common and benign pelvic condition, endometriosis can rarely manifest in the inguinal region, within the canal of Nuck. The treating physician should be cognizant of Nuck canal endometriosis, especially in young female patients presenting with an irreducible mass in the inguinal region and associated cyclic pain or infertility. Conclusion: When clinically and radiologically suspected, surgical excision is indicated to establish the diagnosis, provide symptomatic relief and guide further decision making.

Background: Malignant Brenner tumors are rare ovarian tumors, accounting for less than 1% of malignant ovarian neoplasms. The aim of this manuscript is to systematically review the current literature concerning malignant Brenner tumors. Methods: We searched three medical databases (PubMed, Scopus, and Web of Science) for relevant articles published until 15 September 2023. Results: After applying inclusion and exclusion criteria, 48 manuscripts describing 115 cases were included in this study from the English literature. Conclusions: We analyzed the demographic, clinical, pathological, and oncological characteristics of 115 patients with malignant Brenner tumors. The statistical analysis showed that recurrence was marginally statistically significantly related to tumor stage and was more common in patients with ascites and in women with abnormal CA-125 levels; patients that were treated with lymphadenectomy had better disease-specific survival.

Breast carcinoma is the most common cancer in women. Nineteen different subtypes of breast carcinomas are recognized in the current WHO classification of breast tumors. Except for these subtypes, there are a number of carcinomas with special morphologic and immunohistochemical features that are not included in the 5th WHO classification, while others are considered special morphologic patterns of invasive breast carcinoma of no special type. In this manuscript, we systematically review the literature on four different subtypes of invasive breast carcinoma, namely lymphoepithelioma-like breast carcinoma, breast carcinoma with osteoclast-like giant cells, signet-ring breast carcinoma, and metaplastic breast carcinoma with melanocytic differentiation. We describe their clinicopathological characteristics, focusing on the differential diagnosis, treatment, and prognosis.

Epithelioid hemangioendothelioma (EHE) is a rare malignant vascular tumor whose pleural EHE (pEHE) type is even more uncommon, with significant heterogeneity in the clinical behavior as well as challenging diagnosis and treatment decisions. Herein, we present a 74-year-old woman admitted to the hospital with dyspnea, pleural effusion, and refractory euvolemic hyponatremia. Chest computed tomography revealed a large right-side pleural effusion with irregular thickening of the parietal pleura and localized nodularity. Histologic evaluation of the thoracoscopic pleural biopsy tissue was used to confirm the diagnosis of pEHE. Assessment of the volume status suggested that the hyponatremia was attributable to a paraneoplastic endocrine syndrome. Administration of the multitarget tyrosine kinase inhibitor pazopanib helped achieve disease control, with the patient remaining free of symptoms after 16 months of follow-up. This case report adds to the knowledge base of this exceptionally rare condition, highlighting the need for a multidisciplinary approach.

Detection of extranodal extension on histopathology in surgically treated head and neck squamous cell carcinoma indicates poor prognosis. However, there is no consensus on the diagnostic criteria, interpretation, and reporting of histology detected extranodal extension, which has contributed to conflicting evidence in the literature, and likely clinical inconsistency. The Head and Neck Cancer International Group conducted a three-round modified Delphi process with a group of 19 international pathology experts representing 15 national clinical research groups to generate consensus recommendations for histology detected extranodal extension diagnostic criteria. The expert panel strongly agreed on terminology and diagnostic features for histology detected extranodal extension and soft tissue metastasis. Moreover, the panel reached consensus on reporting of histology detected extranodal extension and on nodal sampling. These consensus recommendations, endorsed by 19 organisations representing 34 countries, are a crucial development towards standardised diagnosis and reporting of histology detected extranodal extension, and more accurate data collection and analysis.

Purpose: We conducted a phase II randomized noncomparative window of opportunity (WOO) trial to evaluate the inhibition of cellular proliferation and the modulation of immune microenvironment after treatment with olaparib alone or in combination with cisplatin or durvalumab in patients with operable head and neck squamous cell carcinoma (HNSCC). Experimental Design: Forty-one patients with HNSCC were randomized to cisplatin plus olaparib (arm A), olaparib alone (arm B), no treatment (arm C) or durvalumab plus olaparib (arm D). The primary endpoint was to evaluate the percentage of patients in each arm that achieved a reduction of at least 25% in Ki67. Secondary endpoints included objective response rate (ORR), safety, and pathologic complete response (pCR) rate. Paired baseline and resection tumor biopsies and blood samples were evaluated for prespecified biomarkers. Results: A decrease in Ki67 of at least 25% was observed in 44.8% of treated patients, as measured by quantitative immunofluorescence. The ORR among treated patients was 12.1%. pCR was observed in 2 patients. Two serious adverse events occurred in 2 patients. Programmed death ligand 1 (PD-L1) levels [combined positive score (CPS)] were significantly higher after treatment in arms A and D. Expression of CD163 and colony-stimulating factor 1 receptor (CSF1R) genes, markers of M2 macrophages, increased significantly posttreatment whereas the expression of CD80, a marker of M1 macrophages, decreased. Conclusion: Preoperative olaparib with cisplatin or alone or with durvalumab was safe in the preoperative setting and led to decrease in Ki67 of at least 25% in 44.8% of treated patients. Olaparib-based treatment modulates the tumor microenvironment leading to upregulation of PD-L1 and induction of protumor features of macrophages. Significance: HNSCC is characterized by defective DNA repair pathways and immunosuppressive tumor microenvironment. PARP inhibitors, which promote DNA damage and “reset” the inflammatory tumor microenvironment, can establish an effective antitumor response. This phase II WOO trial in HNSCC demonstrated the immunomodulatory effects of PARP inhibitor–induced DNA damage. In this chemo-naïve population, PARP inhibitor–based treatment, reduced tumor cell proliferation and modulated tumor microenvironment. After olaparib upregulation of PD-L1 and macrophages, suggests that combinatorial treatment might be beneficial. Synopsis: Our WOO study demonstrates that preoperative olaparib results in a reduction in Ki67, upregulation of PD-L1 CPS, and induction of protumor features of macrophages in HNSCC.

Supplementary Figure 6. Posttreatment CD163 (by QIF) increase in three patients with concurrent increase in the CD163 transcripts. Representative images from a patient’s tumor tissue sample showing pre and post treatment (Durvalumab-Olaparib Arm) CD163 expression. Nuclei (blue), Cytokeratin (green), CD163 (yellow) and CSF1R (red).

BACKGROUND/AIM:Adenoid cystic carcinoma (ACC) is an aggressive neoplasm even though it has low-grade histological appearance and slow growth. The aim of this study was to identify the immunohistochemical and molecular characteristics of ACC, as well as their correlation with the clinical course of patients.PATIENTS AND METHODS:This is a retrospective multicenter analysis. We included 50 patients diagnosed with ACC in the head and neck between 2000 and 2021. The expression of MYB proto-oncogene transcription factor (MYB), neurotrophic tyrosine kinase receptor (NTRK), human epidermal receptor-2 (HER-2), and Ki-67 was examined through immunohistochemistry (IHC) and fluorescence in situ hybridization (FISH). We also performed a clinical follow-up of the patients.RESULTS:The median age of the patients was 58.5 years; moreover, 54% of the patients were male. Compared with female patients, male patients were at a higher risk of both recurrence and death. No HER-2-positive cases were revealed. MYB expression was positive in 28 (56%) cases. However, MYB expression did not significantly affect survival. NTRK expression was positive in eight (16%) cases. NTRK-positive patients had worse overall survival (OS) than NTRK-negative patients (p=0.0246). Additionally, the percentage of NTRK-stained cells was negatively correlated with disease-free survival (p=0.0016) and OS (p=0.0027).CONCLUSION:There was no correlation between MYB positivity and survival. Contrarily, NTRK-positive patients had worse survival, indicating that NTRK is a negative prognostic factor. Tropomyosin receptor kinase inhibitors can be used to treat these patients. Furthermore, MYB-targeted inhibitors are promising therapeutic agents.

Perivascular epithelioid cell tumors are very rare mesenchymal neoplasms arising in various locations, such as the female genital tract, kidney, lung, prostate, bladder, pancreas, soft tissues, and bone. They possess a unique immunophenotype, co-expressing myogenic and melanocytic markers; molecular findings include mutations of tuberous sclerosis complex and translocations of transcription factor E3, a member of the microphthalmia transcription factor gene family. We herewith report a uterine collision tumor consisting of a perivascular epithelioid cell tumor and a moderately differentiated endometrial endometrioid carcinoma in a patient with genetically proven tuberous sclerosis; two leiomyomas were also found in contact with the tumor. Although two such cases one with a benign and another with a malignant perivascular epithelioid cell tumor have previously been reported, ours is, to our knowledge, the first reported in a tuberous sclerosis patient.