Multidimensional Characterization of Tumor–Immune Architecture Reveals Clinically Relevant Classic Hodgkin Lymphoma Subtypes

The tissue architecture of classic Hodgkin lymphoma (cHL) is unique among cancers and is characterized by rare malignant Hodgkin and Reed-Sternberg cells that coevolve with a complex ecosystem of immune cells in the tumor microenvironment (TME). The lack of a comprehensive systems-level interrogation has hindered the description of disease heterogeneity and clinically relevant molecular subtypes. In this study, we employed an integrative, multimodal approach to characterize cHL tumors using malignant cell sequencing, spatial transcriptomics, and imaging mass cytometry. We identified four molecular subtypes (CST, CN913, STB, and CN2P), each characterized by distinct clinical features, mutational patterns, malignant cell gene expression profiles, and spatial architecture involving immune cell populations. Functional modeling of CSF2RB mutations, a characteristic feature of the CST subtype, revealed dysregulated oncogenic signaling and unique TME cross-talk. These findings highlight the significance of multidimensional profiling in elucidating patterns of molecular alterations that drive immune ecosystems and underlie therapeutically exploitable vulnerabilities. SIGNIFICANCE: Our systematic and comprehensive genomic and spatially resolved profiling of Hodgkin lymphoma revealed a paradigmatic link between a genetic disease subtype and TME composition and function. Insights into mutation-driven mechanisms of deregulated cytokine signaling will pave the way for therapeutic interventions that interfere with oncogenic signaling and cellular ecosystems in cancer.