Objectives Timely access to pathology reports has increased the need for clear patient-facing explanations. We evaluated whether large language model (LLM)-generated responses to pathology report questions from patients are comparable in quality to explanations written by pathologists and assessed how LLM configuration influences performance.Methods Sixty-five anonymized real-world patient questions from an online pathology education platform were answered using 5 LLM configurations varying by model architecture, prompting strategy, and retrieval-augmented generation. Responses were evaluated using a structured rubric that assessed accuracy, relevance, clarity, empathy, and safety; they were compared using pairwise arena testing with Bradley-Terry modeling to rank performance.Results Across rubric domains, LLM responses demonstrated performance comparable to pathologist explanations, with 1 configuration meeting noninferiority criteria. Pairwise arena comparisons indicated that the configuration parameters strongly influenced performance, with both model size and retrieval augmentation associated with improved response preference. The highest-performing configuration combined a larger model with retrieval from a curated pathology knowledge base and was strongly preferred over pathologist-written responses.Conclusions Carefully configured LLM systems can generate patient-facing explanations of pathology reports comparable in quality to pathologist-written explanations. Prompting strategy, model size, and retrieval integration were associated with performance differences, underscoring the importance of system configuration in developing LLM-based tools to expand access to understandable pathology information.

Anaplastic thyroid cancer (ATC) is highly lethal. Although patients with the BRAFV600E alteration respond to the type I RAF inhibitor (RAFi) dabrafenib with trametinib, most rapidly develop adaptive or acquired resistance. Here, multi-region whole-genome, high-coverage whole-exome, and single-nuclei RNA sequencing of tumors from ATC patients undergoing type I RAFi and MEKi therapy reveals that reactivation of the mitogen-activated protein kinase (MAPK) pathway, along with immunosuppressive macrophage proliferation, may underlie the development of acquired resistance. Screening of several RAFi reveals that ATC cell lines are exquisitely sensitive to the type II RAFi naporafenib, which inhibits EphA2-mediated MAPK signaling. Further, naporafenib and trametinib overcome both innate and acquired treatment resistance to dabrafenib and trametinib in ATC cell lines and patient-derived xenograft models. Finally, we describe a mechanism of acquired resistance to naporafenib through compensatory mutations in MAST1. Taken together, our work rationalizes the clinical investigation of type II RAFi in the setting of thyroid cancer.

Abstract Cancer cell dormancy and the resultant resistance to conventional therapies present significant challenges for the successful treatment of high-grade serous ovarian cancer (HGSC). We used genome wide, and specialized sgRNA, libraries in CRISPR-based screens to identify critical cell survival mechanisms in dormancy and metastasis. Our findings demonstrate that low expression Wnt ligands WNT8B and WNT9B are essential for sustaining cell survival during prolonged dormant spheroid culture conditions. These Wnt ligands utilize non-canonical signaling to activate expression of stem cell genes such as ALDH1A1 , CD44 and others during spheroid dormancy. The loss of WNT8B and WNT9B reduced survival of xenografted ovarian cancer cells during early dissemination of disease that extended survival. Furthermore, treatment of WNT8B/9B deficient xenografts with carboplatin demonstrated increased sensitivity that further reduced dissemination and extended survival. These findings reveal that rare Wnt ligands can possess outsized functions in cancer pathogenesis and offer new avenues for improving treatment outcomes for HGSC through their inhibition.

Introduction:HLA-G is a non-classical major histocompatibility complex class I molecule with potent immunosuppressive activity and is increasingly recognized as an immune-checkpoint axis in cancer. Its prognostic significance in non-small cell lung cancer (NSCLC), particularly in relation to PD-L1 expression and CD8+ tumor-infiltrating lymphocytes (TILs), remains incompletely defined. Methods:We retrospectively analyzed 314 surgically resected NSCLCs assembled in tissue microarrays and stained for HLA-G, PD-L1, and CD8. HLA-G and PD-L1 were scored as positive when ≥1% of tumor cells showed membranous staining, whereas CD8+ TIL density was digitally quantified and dichotomized using the cohort median (≥575 cells/mm²). Associations with clinicopathological variables and outcomes were assessed by Kaplan-Meier analysis and multivariable Cox regression. Results:HLA-G was expressed in 50 of 314 tumors (16%), PD-L1 in 106 of 314 (33.8%), and high CD8 density in 160 of 314 (51%). In HLA-G-negative tumors, high CD8+ TIL density was associated with significantly prolonged disease-free survival (DFS) and overall survival (OS). In the overall cohort, the combined HLA-G-negative/CD8-high phenotype retained independent favorable prognostic significance for both DFS and OS. By contrast, CD8 density did not significantly stratify outcome in HLA-G-positive tumors, although these subgroup analyses were limited by small sample size. Discussion:In combined biomarker analyses, the favorable prognostic effect of CD8+ TILs in PD-L1-negative tumors was maintained only when HLA-G was also absent. Within the PD-L1-positive/HLA-G-negative subgroup, high CD8 density was independently associated with improved DFS but not OS. Integrating HLA-G with PD-L1 and CD8 assessment may refine prognostic stratification and help identify patients who could benefit from HLA-G-targeted strategies, alone or in combination with PD-1/PD-L1 blockade.

Eye tracking technology offers the opportunity to gain insight into the mind of the pathologist during tissue assessment. Prior work has focused on relating regions of pathologist's fixations on tissue to areas of diagnostic importance, however the fine movements of the eye during a fixation, known as fixational eye movements (FEMs), have yet to be explored in the context of digital pathology. In this work, we sought to determine whether a pathologist's FEMs could be used to distinguish mitotic figures from mitotic figure confounders, with above chance performance. From a single pathologist assessing a whole slide image, eye tracking was used to collect 316 mitotic figure and 251 mitotic figure confounder FEM clusters. Radiomics and convolutional neural network (CNN) classifier approaches were investigated for FEM cluster classification. Exploring the parameter and hyperparameter spaces of both approaches, both radiomics and CNN classifiers demonstrated above chance classification performance (median area under the receiver operating characteristic curve; AUC >= 0.6), with the highest performances having AUCs of 0.68 and 0.71 for the radiomics and CNN approaches, respectively. A secondary analysis of bone marrow cells from two trained professionals was performed and demonstrated that the same radiomic machine learning techniques could also distinguish bone marrow cells with good accuracy (median accuracy of 0.85 and 0.64), using only FEM clusters as input. This work demonstrates that a trained professional ' s FEMs can be used to distinguish various cells types in digital pathology and shows promise that a pathologist ' s fixations during diagnostic assessment encode retrievable information about the object being observed, supporting future investment in this new avenue of research in human-machine interfaces for digital pathology, including the potential for rapid labeling of tissue structures based on inference from eye gaze data.

Amiodarone is an effective and commonly used anti-arrhythmic drug. The radiographic presentations of amiodarone-induced pulmonary toxicity (AIPT) include a broad spectrum of patterns. Three main clinical-radiographic presentations are described with AIPT: isolated reduction of diffusing lung capacity with no clinical implications; alveolar-interstitial pneumonia, which usually requires amiodarone discontinuation and corticosteroid treatment; and acute respiratory distress syndrome, which can obviously be life-threatening. We report 6 cases that depict the described range of clinical and radiographic presentations. The analysis of the cases and of the published review suggests that the computed tomography scan pattern on presentation predicts well the clinical course. The correlation between radiographic and clinical findings can therefore guide the clinician in the management of AIPT.

[This corrects the article DOI: 10.3389/fimmu.2026.1732852.].

CONTEXT.—:Social media is a powerful tool in pathology education and professional networking that connects pathologists and pathology trainees from around the world. Twitter (X) appears to be the most popular social media platform pathologists use to share pathology-related content and connect with other pathologists. Although there has been some published research on pathology-related activity on Twitter during short time frames, to date there has not been published research examining pathology-related Twitter activity in totality from its earliest days of activity to recently. OBJECTIVE.—:To comprehensively evaluate the use of pathology on Twitter (X) during the last 10 years. DESIGN.—:Pathology-related tweets were systematically scraped from Twitter from January 2012 to January 2023 using pathology hashtags as a surrogate measure for all pathology content on Twitter. COVID-related tweets were approximated by tweets containing the term "COVID." RESULTS.—:There were 591 812 unique pathology-related tweets identified during the time period, with #pathology being the most common hashtag used and #PathTwitter becoming more popular since 2020. There has been positive annual growth of pathology Twitter, with peaks in use during major pathology conferences. During the initial phases of the COVID-19 pandemic, a sustained increase in pathology tweets was observed. CONCLUSIONS.—:Pathology Twitter has grown during the last 10 years and has become increasingly popular for pathology education and networking. With the changing landscape of social media platforms, this study provides an understanding of how pathology medical education and professional networking uses of social media happen and evolve over time.

SUMMARY:NanoString's GeoMx Digital Spatial Profiling platform enables researchers to elucidate spatial transcriptomic profiles of distinct cellular microenvironments in human and mouse formalin fixed paraffin embedded tissue. To date, there is no free, open source, interactive application to facilitate data analysis with the latest tested methods. We created shinyDSP, a R shiny application that guides users to perform quality control, normalization, and differential gene expression analysis of GeoMx DSP data. It includes various user-provided customization options to meet individual aesthetic choices and requirements. AVAILABILITY AND IMPLEMENTATION:The release and development versions of shinyDSP are available on Bioconductor under the MIT license (https://www.bioconductor.org/packages/release/bioc/html/shinyDSP.html) and Github (https://github.com/kimsjune/shinyDSP).

CONTEXT.—:Digital pathology requires pathologists to assess tissue digitally rather than on an analog microscope, which has been the mainstay tool for tissue assessment for more than a century. The impact of different digital interaction configurations on pathologists' performance is not well understood. This work focuses on the impact of the display window size for diagnostic assessment. OBJECTIVE.—:To determine the effect of digital image viewer window size on pathologists' diagnostic performance when searching for tumors in lymph nodes while under a time limit. DESIGN.—:Six pathologists assessed 8 breast lymph node whole slide images using 4 digital image viewer window sizes (8, 14, 24, and 32 inches) for tumors in lymph nodes while under a time limit. Eye-gaze data were collected. Pathologists were subsequently asked to rate their preference of window sizes. RESULTS.—:The fraction of window not covered with foveated vision was significantly associated with window size ranging from 43% for 32 inches to 5% for 8 inches (P < .001). There was no statistically significant relationship between the number of false negatives or assessment time and window size (P = .21 and P = .28, respectively). The distance traversed per panning instance ranged from 301 pixels for 32-inch to 193 pixels for 8-inch windows (P = .002). All pathologists preferred the largest window size as it provided more context for diagnostic assessment. CONCLUSIONS.—:Window size does not significantly affect pathologists' diagnostic performance when searching for tumors in lymph nodes. However, pathologists adapted their slide navigation approach to accommodate the amount of context the window size permitted.

BACKGROUND:First described in 1972, idiopathic subglottic stenosis (iSGS) is a serious chronic orphan disease characterised by recurrent scarring of the subglottis. Although the cause is unknown, iSGS is almost exclusively restricted to Caucasian females typically in their fourth to sixth decade. However, given its rare incidence (1:400,000), understanding the clinical trajectory and molecular factors associated with iSGS disease development and prognosis has been difficult. In the current study we sought to unravel the pathogenesis of iSGS at the clinical, transcriptional, and genetic level in a prospective cohort. METHODS:We prospectively enrolled 126 patients with iSGS, 104 controls, and 13 patients with traumatic SGS. Within this cohort, we profiled 114 human epiglottis and 121 human subglottis biopsies across three different conditions: control, iSGS, and intubation-related traumatic stenosis using bulk and single nucleus RNA-sequencing. Whole exome sequencing for germline variants was performed for 70 controls and 75 patients with iSGS. FINDINGS:Patients with iSGS received a median number of five (range 0-18) surgical dilations at a rate of 1.031 dilations (range: 0.12-6.2) per year. Older age at diagnosis and higher Cotton-Myers grade were associated with increased number of surgical dilations over time. Cohort-level bulk transcriptomics found that iSGS pathology was restricted within the subglottis and did not affect anatomically adjacent epiglottis, opposite to previous hypotheses. We further identified cellular subsets associated with iSGS prognosis and severity. Finally, patients with iSGS exhibit lower testosterone predicted using a polygenic score. INTERPRETATION:Together, our data refines our understanding of laryngeal biology and provides insights into the clinical trajectory of subglottic stenoses. Future research should explore the role of testosterone in the development of iSGS. FUNDING:This study was funded by a grant from the American Laryngology Association (#1082), an Academic Medical Organization of Southwestern Ontario innovation fund grant (INN21-016), grant support from the Departments of Otolaryngology-Head and Neck Surgery at University of Toronto and Western University. ACN was supported by the Wolfe Surgical Research Professorship in the Biology of Head and Neck Cancers Fund. PYFZ was supported by a Vanier Canada Graduate Scholarship and PSI foundation fellowship.

Neoadjuvant chemoradiation therapy (NCRT) is an underutilized treatment in breast cancer but may improve outcomes by impacting the tumor immune microenvironment. The aim of this study was to evaluate NCRT’s impact on recurrence and the role of tumor-infiltrating lymphocytes (TILs) in treatment response. We hypothesized that NCRT reduces recurrence by upregulating TILs. Patients with locally advanced breast cancer (LABC) were treated with NCRT. Stage IIB to III patients with any molecular subtypes were eligible. The patients were matched for age, stage, and molecular subtype by a propensity score to a concurrent cohort receiving standard neoadjuvant chemotherapy (NCT) followed by adjuvant radiation. The objective of this study was to assess the patients in terms of the pathological complete response (pCR), TIL counts prior to and following treatment, and locoregional recurrence. The median follow-up was 7.2 years. Thirty NCRT patients were successfully matched 1:3 to ninety NCT patients. The NCRT cohort had no regional and locoregional recurrences (p = 0.036, (hazard ratio) HR [0.25], 95% confidence interval (CI) [0.06–0.94] and p = 0.013, HR [0.25], 95% CI [0.08–0.76], respectively), compared to 17.8% of the NCT cohort. The NCRT group had significantly more pCRs, and TILs were increased in the post-treatment pCR specimens. NCRT can improve outcomes in LABC patients, with a higher pCR and significantly lower locoregional recurrence/higher recurrence-free survival. Further trials are needed to evaluate the role of NCRT in all breast cancer patients.

OBJECTIVE:Idiopathic subglottic stenosis (iSGS) is a rare disease characterized by narrowing of the upper airway and affects near-exclusively females. Patients often experience recurrent disease and require repeated surgical dilations. The pathophysiology underlying the broad spectrum of disease severity within iSGS remains unknown. In the current study, we sought to identify transcriptomic differences between iSGS patients with markedly different recurrence rates. METHODS:Prospectively collected clinical and bulk RNA sequencing data from subglottic tissues of 56 female iSGS patients with 1-4 years of follow-up were analyzed. DESeq2 was used to perform differential expression analysis, comparing samples from the highest (1.19-1.87 dilations/year) versus the lowest (0.30-0.65 dilations/year) quartile of surgical dilation rate (i.e., high vs. low recurrence groups). RESULTS:In total, 220 genes were significantly differentially expressed between the high and low recurrence groups (adjusted p < 0.1 and log2 fold change > |1|). Pathway enrichment analyses showed that the high recurrence group had significantly increased expression of genes involved in adaptive immune responses (e.g., immunoglobulin subunit genes) and extracellular matrix organization (e.g., COMP, NID2) (adjusted p < 0.1). In contrast, the low recurrence group had significantly increased expression of genes involved in cilia structure and function (e.g., CFAP43, DNAI2) (adjusted p < 0.1), suggesting a relatively increased abundance of respiratory cilia. CONCLUSION:Transcriptomic profiling suggests that lower recurrence rates in iSGS are associated with retention of respiratory cilia, while adaptive immune responses and increased extracellular matrix deposition are present in those with higher recurrence rates. These results hold promise for the development of prognostic markers and identification of therapeutic targets for iSGS.

Brentuximab vedotin for scleroderma skin

OBJECTIVE:We explored the efficacy and safety of brentuximab vedotin, a chimeric anti-CD30 antibody drug conjugate, in patients with severe active diffuse cutaneous systemic sclerosis (dcSSc). METHODS:This phase II proof-of-concept, single centre, open-label, single arm, investigator-initiated trial included patients ≥18 years, with dcSSc, modified Rodnan skin score (mRSS) ≥15 with <5 years since the first non-Raynaud's symptom and/or skin worsening despite immunosuppression who were treated with intravenous brentuximab vedotin 0.6 mg/kg q3 weeks for 45 weeks. The primary end point was a decrease in mRSS of ≥8 points at 48 weeks. RESULTS:Eleven patients were treated with brentuximab vedotin, with nine completing the study. The mean mRSS reduction at week 48 was 11.3 (95% CI 6.9, 15.8; P = 0.001), meeting the primary end point in the intention to treat analysis (7/11 had a decrease in mRSS ≥8). The % forced vital capacity increased by 7.8% (12.5). The Composite Response Index in dcSSc (CRISS) suggested a beneficial treatment effect (86% ≥0.6). Most adverse events were mild. No SAEs were attributed to brentuximab vedotin. CONCLUSION:In dcSSc, brentuximab vedotin improved skin and FVC without safety concerns. A placebo-controlled trial is warranted to corroborate these initial findings. TRIAL REGISTRATION:ClinicalTrials.gov, http://clinicaltrials.gov, NCT03198689.

Rationale Fibrosing interstitial lung diseases (ILDs), including idiopathic pulmonary fibrosis (IPF), non-specific interstitial pneumonia (NSIP), and chronic hypersensitivity pneumonitis (CHP), are characterized by progressive lung scarring. While single-cell RNA-seq (scRNA-seq) has provided insights into the cellular landscape of normal and diseased lungs, a comprehensive spatial map of these diseases remains lacking. Thus, we sought to fill this gap by generating a spatial transcriptomic atlas of fibrosing ILDs. Methods We used formalin-fixed, paraffin-embedded surgical lung biopsies from treatment-naïve patients with IPF (n=10), NSIP (n=8), CHP (n=10), or unclassified ILDs (n=17). Spatial transcriptomics was performed with the Visium platform (10X Genomics) to capture a 6.5 mm by 6.5 mm square area. After performing quality control, we integrated single-cell annotations from the Integrated Human Lung Atlas and used “cell2location” to map cell type proportions on tissues. We used non-negative matrix factorization (NMF) and ‘scanpy’ to identify co-localizing cell types and differentially expressed genes between groups, respectively. Results Cell type mapping was consistent with histological findings, as we identified known marker genes for each cell type among the top differentially expressed genes. We also identified genes such as DDIT4 and TSC22D3, SFTPC, and TIMP1 and TAGLN that were strongly associated with fibroblasts in IPF, CHP and NSIP, respectively. While no cell types were specifically enriched in a particular ILD subtype, NMF (R=7) revealed differential co-localization patterns across conditions. For example, in CHP, AT1 and AT2 cells formed two distinct factors, a pattern not observed in IPF or NSIP. Conclusion This ongoing study provides the first large scale spatial transcriptomic atlas of fibrosing ILDs. We successfully integrated scRNA-seq data to predict cell type proportions within spatial contexts. Additional plans include the generation of an algorithm to reclassify unclassifiable cases and a correlation analysis between clinical outcomes and spatial gene expression patterns, independent of the ILD subtype.