Biology subjects

Madore, J.

Publications and source records attributed to Madore, J..

2 recordsLinked to original sources

bioRxiv2026

An open multimodal spatial resource integrating same-tissue transcriptomics, proteomics, and histology

Spatial transcriptomic and proteomic technologies provide complementary insights into tissue organisation, cellular phenotype and function, yet integrating these modalities on the same tissue section remains technically challenging. Sequential workflows must preserve RNA integrity, antigenicity and tissue morphology while maintaining accurate spatial registration. At present, publicly available multimodal datasets suitable for computational method development remain limited. Here, we present a workflow for sequential 10x Genomics Xenium spatial transcriptomics, COMET cyclic immunofluorescence, and haematoxylin and eosin (H&E) histological staining on the same formalin-fixed paraffin-embedded tissue section. We demonstrate this approach across multiple biologically distinct human tissues, including tonsil, hepatocellular adenoma, and matched tumour and non-tumour hepatocellular carcinoma, illustrating the widespread applicability of the workflow beyond a single tissue type. Following image registration, Xenium-derived cell segmentations were applied to protein images to generate integrated single-cell transcriptomic and proteomic measurements for downstream analyses. To facilitate community reuse, we publicly release four representative aligned tissue cores together with transcript coordinates, multiplex protein images, H&E images, cell segmentations, and integrated single-cell datasets. We additionally introduce UnumLocalia, an open-source visualisation and data extraction tool that enables interactive exploration of aligned multimodal images, supports user-defined cell segmentation, and allows export of integrated single-cell data for downstream analyses. Together, this technical protocol, workflow, software, and openly available dataset provide a reusable resource for multimodal spatial biology, supporting advances in biological discovery, computational method development, multimodal data integration, and validation of emerging analytical approaches across complementary spatial technologies.

immunology↗

bioRxiv2024

Type I interferon drives a cellular state inert to TCR-stimulation and could impede effective T-cell differentiation in cancer

Head and neck squamous cell carcinoma (HNSCC) arises from the mucosal epithelium of the oral cavity, pharynx, or larynx and is linked to exposure to classical carcinogens and human papillomavirus (HPV) infection. Due to molecular, immunological, and clinical disparities between HPV+ and HPV-HNSCC, they are recognized as distinct cancer types. While immune checkpoint inhibition (ICI) has demonstrated efficacy in recurrent/metastatic HNSCC, response variability persists irrespective of HPV status. To gain insights into the CD8+ T-cell landscape of HPV-HNSCC, we performed multimodal sequencing (RNA and TCR) of CD8+ tumor-infiltrating lymphocytes (TILs) from treatment-naive HPV-HNSCC patients. Additionally, we subjected cells to ex vivo TCR-stimulation, facilitating the tracing of clonal transcriptomic responses. Our analysis revealed a subset of CD8+ TILs highly enriched for interferon-stimulated genes (ISG), which were found to be clonally related to a subset of granzyme K (GZMK)-expressing cells. Trajectory inference suggests ISG transition via GZMK cells towards terminal effector states. However, unlike GZMK cells, which rapidly an effector-like phenotype in response to TCR stimulation, ISG cells remain transcriptionally inert. Consequently, ISG cells may impede effective T-cell differentiation within the TME. Although, the functional consequences of ISG cells are poorly understood, we revealed that they possess receptors and ligands enabling cell-cell communication networks with key TME immunomodulators such as dendritic cells. Additionally, ISG cells were found to be a core feature across various tumor entities and were specifically enriched within tumor tissue. Thus, our findings illuminate the complexity of T-cell heterogeneity in HPV-HNSCC and reveal an overlooked population of IFN-stimulated CD8+ TILs. Further exploration of their functional significance may offer insights into therapeutic strategies for HPV-HNSCC and other cancer types.

immunology↗