Background: Adaptive immune responses to structural proteins of the virion play a crucial role in protection against coronavirus disease 2019 (COVID-19). We therefore studied T cell responses against multiple SARS-CoV-2 structural proteins in a large cohort using a simple, fast, and high-throughput approach.Methods: An automated interferon gamma release assay (IGRA) for the Nucleocapsid (NC)-, Membrane (M)-, Spike-C-terminus (SCT)-, and N-terminus-protein (SNT)-specific T cell responses was performed using fresh whole blood from study subjects with convalescent, confirmed COVID-19 (n = 177, more than 200 days post infection), exposed household members (n = 145), and unexposed controls (n = 85). SARS-CoV-2-specific antibodies were assessed using Elecsys^® Anti-SARS-CoV-2 (Ro-N-Ig) and Anti-SARS-CoV-2-ELISA (IgG) (EI-S1-IgG).Results: 156 of 177 (88%) previously PCR confirmed cases were still positive by Ro-N-Ig more than 200 days after infection. In T cells, most frequently the M-protein was targeted by 88% seropositive, PCR confirmed cases, followed by SCT (85%), NC (82%), and SNT (73%), whereas each of these antigens was recognized by less than 14% of non-exposed control subjects. Broad targeting of these structural virion proteins was characteristic of convalescent SARS-CoV-2 infection; 68% of all seropositive individuals targeted all four tested antigens. Indeed, anti-NC antibody titer correlated loosely, but significantly with the magnitude and breadth of the SARS-CoV-2-specific T cell response. Age, sex, and body mass index were comparable between the different groups.Conclusion: SARS-CoV-2 seropositivity correlates with broad T cell reactivity of the structural virus proteins at 200 days after infection and beyond. The SARS-CoV-2-IGRA can facilitate large scale determination of SARS-CoV-2-specific T cell responses with high accuracy against multiple targets.

Incidental spinal bone lesions, potential indicators of malignancies, are frequently underreported in abdominal and thoracic CT imaging due to scan focus and diagnostic bias towards patient complaints. Here, we evaluate a deep-learning algorithm (DLA) designed to support radiologists’ reporting of incidental lesions during routine clinical practice. The present study is structured into two phases: unaided and AI-assisted. A total of 32 scans from multiple radiology centers were selected randomly and independently annotated by two experts. The U-Net-like architecture-based DLA used for the AI-assisted phase showed a sensitivity of 75.0% in identifying potentially malignant spinal bone lesions. Six radiologists of varying experience levels participated in this observational study. During routine reporting, the DLA helped improve the radiologists’ sensitivity by 20.8 percentage points. Notably, DLA-generated false-positive predictions did not significantly bias radiologists in their final diagnosis. These observations clearly indicate that using a suitable DLA improves the detection of otherwise missed potentially malignant spinal cases. Our results further emphasize the potential of artificial intelligence as a second reader in the clinical setting.

Spinal bone lesions encompass a wide array of pathologies, spanning from benign abnormalities to aggressive malignancies, such as diffusely localized metastases. Early detection and accurate differentiation of the underlying diseases is crucial for every patient's clinical treatment and outcome, with radiological imaging being a core element in the diagnostic pathway. Across numerous pathologies and imaging techniques, deep learning (DL) models are progressively considered a valuable resource in the clinical setting. This review describes not only the diagnostic performance of these models and the differing approaches in the field of spinal bone malignancy recognition, but also the lack of standardized methodology and reporting that we believe is currently hampering this newly founded area of research. In line with their established and reliable role in lesion detection, this publication focuses on both computed tomography and magnetic resonance imaging, as well as various derivative modalities (i.e. SPECT). After conducting a systematic literature search and subsequent analysis for applicability and quality using a modified QUADAS-2 scoring system, we confirmed that most of the 14 identified studies were plagued by major limitations, such as insufficient reporting of model statistics and data acquisition, a lacking external validation dataset, and potentially biased annotation. Although we experienced these limitations, we nonetheless conclude that the potential of these methods shines through in the presented results. These findings underline the need for more stringent quality controls in DL studies, as well as model development to afford increased insight and progress in this promising novel field.

The tumor microenvironment (TME) actively contributes to pancreatic ductal adenocarcinoma (PDAC) pathogenesis through dynamic bidirectional tumor–stroma interactions. In this study, we demonstrated that ATM-deficient tumor epithelium reprograms the TME in a genotype-specific manner to enhance cancer aggressiveness. In genetically engineered mouse models, pancreatic stellate cell and cancer-associated fibroblast (CAF) coculture systems, single-nucleus multiomics, and human PDAC models, tumoral loss of ATM serine/threonine kinase drove CAFs toward αSMA+ myofibroblastic (myCAF) differentiation, independently of p53 status. The myCAFs, in turn, promoted cancer aggressiveness and chemoresistance. Mechanistically, ATM deficiency increased reactive oxygen species and contractility signaling, enhancing TGFβ1 secretion. Pharmacologic TGFβ inhibition reversed myCAF differentiation, sensitized tumors to chemotherapy, and impaired tumor progression in both murine and human ATM-null models. These findings reveal that ATM-deficient tumors shape a cancer-promoting niche via TGFβ signaling and identify dual targeting of intrinsic and extrinsic vulnerabilities as a promising precision oncology strategy. Significance: TGF-β-driven myofibroblastic stromal differentiation in ATM-deficient pancreatic cancer generates a genotype-specific tumor microenvironment, providing a targetable axis and highlighting the need to integrate epithelial genotype and stromal context in pancreatic cancer therapy.

ROS measurements, cytoskeletal marker expression, migration assays, and PSC differentiation analyses following genetic and pharmacologic perturbation of ROS and cytoskeletal pathways in ATM-deficient tumor models.