Citations

Context.— The Goldman criteria are widely used to categorize diagnostic discrepancies in autopsy reports for hospital quality assurance. Despite their adoption, little is known about how consistently they are applied across institutions. Objective.— To evaluate the interrater reliability among academic pathologists applying the Goldman criteria to classify missed diagnoses in autopsy cases. Design.— Thirty autopsy cases from 2 academic hospitals were reviewed by 5 experienced autopsy pathologists. Each reviewer independently assigned Goldman classification scores (class I–IV) to missed diagnoses based on the original 1983 definitions. Consensus was defined as at least 60% agreement (≥3 of 5 raters). Fleiss κ and Gwet AC1 were calculated to assess interrater reliability. Results.— A total of 196 missed diagnoses were identified by at least 1 rater. Only 26% (51 of 196) achieved a consensus Goldman classification, and just 4% (8 of 196) had unanimous classification agreement. Most diagnoses (65%; 127 of 196) were not identified by at least 3 raters. Among consensus classification ratings, 49% (25 of 51) were class IV. Fleiss κ was 0.176 (slight agreement) while Gwet AC1 was 0.390 (fair agreement). Collapsing major (classes I–II) and minor (classes III–IV) categories modestly improved agreement. Conclusions.— Interrater agreement in applying the Goldman criteria is only fair to moderate, suggesting significant subjectivity in their application. As the criteria are used for institutional comparison and quality metrics, efforts to standardize definitions and provide reviewer training are warranted to improve reproducibility and utility.

Establishes a robust long-term human adipocyte organoid culture system maintained for over 90 days. Recapitulates key features of OSA-related intermittent hypoxia in human adipocytes in vitro. Demonstrates IH-induced adipocyte aging, including ultrastructural damage and epigenetic reprogramming. Identifies impaired adipogenesis and insulin signaling under intermittent hypoxia. Provides a physiologically relevant platform for mechanistic studies and drug screening.

Invasive lobular breast carcinoma (ILC), the most common special histological subtype of breast cancer, is characterized by nearly universal expression of estrogen receptor alpha (ER) and unique sites of metastases, neither of which is fully recapitulated by genetically engineered mouse models. Using reporter-labeled ILC mouse xenografts, herein we used mammary fat pad, tail vein and intracardiac orthotopic growth to analyze spontaneous and experimental metastasis and gene expression. We observed ER-positive primary tumors with single-file histology and collagen deposition, and spontaneous metastasis from the mammary fat pad to bones, ovaries, and brain including the leptomeninges, thereby closely mirroring the growth and metastatic spread of human ILC. Brain metastases showed strong ER staining, confirmed by sequencing analyses which identified estrogen signaling as top activated pathway, and the lesions exhibited robust response to endocrine therapy. In summary, we report endocrine responsive mammary fat pad, tail vein and intracardiac xenografts that faithfully demonstrate unique ILC features and can serve as invaluable pre-clinical translational platforms for validating candidate ILC genetic drivers and testing novel therapeutics.

ABSTRACT Lactylation, a protein post-translational modification driven by lactate, plays crucial roles in various physiological and pathological processes. However, the function and underlying mechanisms of lactylation in infection and pathogenesis of influenza A virus (IAV) remain poorly understood. Here, we demonstrated that sirtuin 1 (SIRT1) suppressed IAV replication by mediating delactylation of viral proteins. We found that IAV infection significantly downregulated the expression of SIRT1. Functional studies revealed that depletion of SIRT1 markedly enhanced replication of IAV, whereas overexpression of SIRT1 attenuated the viral replication. Mechanistically, we observed that SIRT1 interacted with IAV NP, PA, PB1, and PB2 proteins, components of the viral ribonucleoprotein (vRNP) complex, and restrained the activity of the vRNP complex, thereby repressing transcription and replication of the IAV genome. Furthermore, we identified that viral NP, PA, PB1, and PB2 proteins underwent lactylation during IAV infection, and SIRT1 facilitated the delactylation of NP, PA, and PB2. Loss of SIRT1 enhanced the lactylation of NP, PA, and PB2, while overexpression of SIRT1 impaired lactylation of these viral proteins during IAV infection. Moreover, inhibition or depletion of lactate dehydrogenase A (LDHA) robustly reduced the lactylation of NP, PA, and PB2, thereby abolishing the promotion of IAV replication caused by SIRT1 deficiency, suggesting that SIRT1 repressed IAV replication via delactylation of viral NP, PA, and PB2 proteins. Together, these results uncover a mechanism that SIRT1 suppresses IAV replication via delactylation of the vRNP components and elicit a critical role for the lactylation of viral proteins in regulating IAV infection and pathogenesis. IMPORTANCE Lactylation plays vital roles in diverse pathological processes. However, the relationship between lactylation and influenza A virus (IAV) infection remains largely unexplored. Particularly, little information is available about the lactylation of viral proteins and their roles in the IAV pathogenesis. Here, viral proteins (NP, PA, PB1, and PB2) were identified to become lactylated during IAV infection. SIRT1 interacted with and promoted the delactylation of viral NP, PA, and PB2 proteins, thereby restraining the vRNP activity and inhibiting the viral transcription and replication. Interestingly, the expression of SIRT1 decreased following IAV infection, suggesting that IAV had evolved a mechanism to downregulate the SIRT1 expression, thereby ensuring appropriate lactylation of viral proteins and facilitating the IAV replication. Together, these findings reveal a critical role for the lactylation of viral proteins in regulating the replication of IAV and provide an important insight into the complicated interplay between the host and IAV.

Cellular and molecular heterogeneity in the liver has been increasingly recognized to drive liver fibrosis progression, but the particular events that occur initially in response to liver injury and trigger immune cell recruitment remain unclear. Here, we identify epigenetically aberrant liver sinusoidal endothelial cells (LSECs) as key players in this process. Mechanistically, the epigenetic readers like bromodomain-containing protein 4–dependent (BRD4-dependent) super enhancers (SEs) activate proinflammatory genes, including promyelocytic leukemia (PML). The PML protein, in turn, binds BRD4 and amplifies proinflammatory angiocrine signaling through phase separation–dependent SE activation via PML/BRD4 condensate formation. In mouse models, LSEC-specific depletion of the PML/BRD4 complex mitigates liver inflammation and fibrosis. Single-cell RNA-seq reveals that epigenetically aberrant LSECs exhibit a reprogrammed proinflammatory angiocrine landscape in mouse fibrotic livers. TIMP1 + LSECs promote the recruitment of CD63 + monocyte–derived macrophages (MoMFs) during liver fibrosis progression. Thereby, PML/BRD4 in LSECs governs inflammatory immune cell recruitment in liver fibrosis. Pharmacological BRD4 inhibition or epigenetic PML-SE repression alleviates liver inflammation and fibrosis. In conclusion, PML/BRD4-mediated SE activation via phase separation drives proinflammatory angiocrine signaling in LSECs, initiating the inflammatory cascade and subsequent immune cell recruitment during liver fibrosis.

Abstract Spatial transcriptomics links gene expression to tissue architecture, but detecting spatially variable genes at atlas scale remains difficult because biologically relevant patterns are multiscale, sparse, and often non-parametric. Here we show that FlashS, a frequency-domain kernel test using random Fourier features, sparse sketching, and a kurtosis-corrected null, retains Gaussian-kernel flexibility at near-linear per-gene cost without permutation. Across 50 benchmark datasets spanning 9 spatial transcriptomics platforms, F lash S achieves the highest ranking accuracy among 14 compared methods while maintaining calibrated inference under simulation and full-atlas permutation. In human heart tissue, it recovers a mitochondrial biogenesis program that co-localizes with ventricular cardiomyocytes. On the Allen Brain MERFISH atlas containing 3.94 million cells, F lash S completes in minutes on a single workstation and separates biological signal from negative-control barcodes even when p-values saturate.

ABSTRACT Malignant peripheral nerve sheath tumors (MPNSTs) are highly aggressive sarcomas with poor prognosis and a strong tendency for metastasis and relapse. Surgical removal remains the mainstay of treatment but is frequently ineffective or impractical. Currently, no effective targeted therapy exists for this type of malignancy. PRMT5 has recently emerged as a promising therapeutic target in various human cancers with MTAP loss, which results in cancer cell dependency on PRMT5 activity. The frequent loss of MTAP in MPNSTs suggests that PRMT5 inhibition is a promising therapeutic option and enables the stratification of cancer patients with few treatment options. We first examined human nerve sheath tumor samples and found that increased PRMT5 expression and activity correlated with MTAP loss in 86.8% (33/38) of MPNSTs and in atypical neurofibromatous neoplasm with uncertain biologic potential (ANNUBP) (5/5). When PRMT5 activity was inhibited genetically and chemically, the cell growth of MTAP-deficient MPNST cell lines was suppressed, but not that of MTAP-proficient MPNST cell lines. Moreover, in the PRMT5-inhibited MTAP-deficient MPNST cell lines, spontaneous DNA damage accumulation was observed following G2/M cell cycle arrest. The DNA replication stress marker RPA32 decreased, and CHK1 was activated early after PRMT5 knockdown, likely contributing to the accumulation of DNA damage. In addition, we combined PRMT5 inhibition with the DNA-damaging agents doxorubicin and gemcitabine, resulting in synergistic effects and increased cancer cell death in MTAP-deficient MPNST cell lines. Together, these findings identify PRMT5 as a compelling therapeutic target in MTAP-deficient MPNSTs. This PRMT5 inhibition strategy has strong translational potential for MPNSTs. GRAPHICAL ABSTRACT

It is well established that many tumor types possess defective autophagic pathways. Several studies have reported that the transmembrane, autophagic lipid scramblase ATG9B is altered in multiple cancers, suggesting that this dysregulation could contribute to oncogenesis. Therefore, the goal of this study was to define the cellular distribution of ATG9B in two different tumor cell types and to provide insights into its cellular function. Surprisingly, we found that ATG9B shows a modest association with autophagic structures and exhibits a unique and prominent localization to mitochondria, in contrast to its related form ATG9A. Upon expression of tagged ATG9B forms, this mitochondrial distribution was accompanied by aberrant changes in mitochondrial morphology as well as a reduction in the mitochondrial membrane potential and the release of mtDNA. Few indicators for ATG9B-dependent mitophagy were noted. Instead, ATG9B overexpression led to pronounced apoptotic cell death as assessed by a variety of indicators. Further, we find that the N-terminal sequence of ATG9B acts as a mitochondrial targeting domain and that expression of this peptide alone can induce apoptotic cell death. These findings provide new insights into a putative cellular localization and function for ATG9B.

Background: Healing of the tendon-bone interface (TBI) after rotator cuff injury is limited, often leading to scar-mediated repair. Decellularized tendon fibrocartilage–bone composite (dTFBC) and bone marrow mesenchymal stem cell sheets (BMSCS) improve repair in preclinical models, but full scaffold encapsulation with mechanical preconditioning remains unexplored. Purpose: To evaluate TBI regeneration using mechanically preconditioned, BMSCS-encapsulated dTFBC scaffolds in a rabbit model. Study Design: Controlled laboratory study. Methods: A total of 48 male New Zealand White rabbits were randomized into 4 groups (n = 12 each): (1) standard repair (control); (2) repair with dTFBC; (3) repair with dTFBC-BMSCS; and (4) repair with mechanically preconditioned dTFBC-BMSCS (mdTFBC-BMSCS). Healing at 12 weeks was assessed via gross observation, histomorphological and immunohistochemical analyses, and biomechanical testing. Results: The mdTFBC-BMSCS group demonstrated a higher ultimate failure load (mean, 203.9 ± 65.6 N) than both the control (118 ± 37.4 N; P = .003) and dTFBC groups (127 ± 44.1 N; P = .008), with no significant difference between the 2 BMSCS-treated groups. Histological analysis revealed enhanced fibrocartilage formation, improved collagen fiber organization, and reduced inflammation infiltration at the TBI in the mdTFBC-BMSCS group. Immunohistochemical analysis showed greater collagen type 2 alpha 1 chain-, interleukin 10-, and arginase 1-positive areas in the mdTFBC-BMSCS group than in the control and dTFBC groups. Conclusion: Mechanically preconditioned, fully BMSCS-encapsulated dTFBC scaffolds promoted TBI regeneration, with enhanced cellular integration, fibrocartilage formation, and favorable biomechanical performance. Clinical Relevance: This biomimetic scaffold strategy may enhance biological healing and reduce the risk of retear after rotator cuff repair.