bims-tumhet Biomed News
on Tumor heterogeneity
Issue of 2025–10–12
four papers selected by
Sergio Marchini, Humanitas Research



  1. Methods Mol Biol. 2026 ;2969 179-193
      Droplet digital PCR (ddPCR) allows highly sensitive detection of minimal amounts of a target of interest. The most common applications of ddPCR have traditionally been detection of genetic variants, copy number variations and gene expression, but the method has great potential also for DNA methylation analyses. We here describe an optimized and standardized protocol for methylation-specific ddPCR analyses, including use of an internal control; the 4Plex, and an automated algorithm for scoring of positive droplets; PoDCall. We also describe a protocol for bisulfite treatment using either of two kits from QIAGEN, utilizing cell-free DNA (cfDNA) or tissue samples as staring material. Implementation of these protocols across laboratories could contribute to more robust and consistent ddPCR methylation results.
    Keywords:  4Plex; Bisulfite treatment; DNA methylation; Droplet digital PCR; Optimization; PoDCall; Protocol; Standardization; cfDNA
    DOI:  https://doi.org/10.1007/978-1-0716-4767-7_12
  2. Nat Commun. 2025 Oct 07. 16(1): 8891
      Mesothelioma is a highly lethal and poorly biologically understood disease which presents diagnostic challenges due to its morphological complexity. This study uses self-supervised AI (Artificial Intelligence) to map the histomorphological landscape of the disease. The resulting atlas consists of recurrent patterns identified from 3446 Hematoxylin and Eosin (H&E) stained images scanned from resected tumour slides. These patterns generate highly interpretable predictions, achieving state-of-the-art performance with 0.65 concordance index (c-index) for outcomes and 88% AUC in subtyping. Their clinical relevance is endorsed by comprehensive human pathological assessment. Furthermore, we characterise the molecular underpinnings of these diverse, meaningful, predictive patterns. Our approach both improves diagnosis and deepens our understanding of mesothelioma biology, highlighting the power of this self-learning method in clinical applications and scientific discovery.
    DOI:  https://doi.org/10.1038/s41467-025-63846-9
  3. Eur J Immunol. 2025 Oct;55(10): e70073
      The immune system is deeply shaped by its anatomical context, with spatial organization emerging as a fundamental principle of immune regulation. Recent advances in spatial omics technologies-encompassing transcriptomics, proteomics, metabolomics, lipidomics, and phosphoproteomics-have revolutionized our ability to study immune processes within intact tissue environments. By preserving spatial coordinates while capturing high-dimensional molecular data, these technologies offer unprecedented insight into how immune cell states and functions are governed by local cues and tissue architecture. In this review, we provide an overview of the major spatial omics platforms, emphasizing methodologies that have gained traction within the immunology community and in our own research. We then illustrate how these tools have begun to elucidate the logic of immune compartmentalization across anatomically complex tissues. While not exhaustive, we highlight selected examples from the intestine, secondary lymphoid organs, and liver to show how spatial omics has uncovered region-specific immune programs, microenvironmental niches, and context-dependent signaling pathways. Together, these studies demonstrate how spatial omics technologies are redefining immunological inquiry-shifting the focus from isolated cell types to their spatially embedded roles in tissue physiology and pathology.
    DOI:  https://doi.org/10.1002/eji.70073
  4. Methods Mol Biol. 2026 ;2969 123-133
      Methods for analyzing circulating tumor DNA (ctDNA) in a fast, quantitative, and reproducible way may help clinical implementation. Analyses of ctDNA can be performed without prior analysis of tumor tissue from patients by using tumor-specific methylations as targets. Digital PCR measurements are quantitative and enable the possibility to monitor dynamics of ctDNA during treatment, which has been shown to correlate with patient outcome. The protocol describes methylation analysis of plasma samples where results are available in a timely manner providing the opportunity to adjust treatment of cancer patients.
    Keywords:  Cancer; Circulating tumor DNA; Droplet digital PCR; Liquid biopsy; Methylation; Plasma
    DOI:  https://doi.org/10.1007/978-1-0716-4767-7_8