Decoding the Heterogeneity of Diffuse Large B-Cell Lymphoma: A Comprehensive Molecular Profiling of B-Cell Lines
Diffuse large B-cell lymphoma (DLBCL) is the most prevalent form of non-Hodgkin lymphoma,exhibiting significant histopathological, molecular and clinical heterogeneity. Advances inclassification integrating phenotypic, genetic, and transcriptomic features have improveddiagnosis and prognosis, guiding therapeutic strategies. However, inconsistencies in themolecular characterization of DLBCL-derived cell lines highlight the need for a standardizedapproach to ensure their reliability as experimental models.This study provides a comprehensive characterization of twenty-nine DLBCL-derived cell lines byintegrating phenotypic, genomic, and transcriptomic analyses to refine classificationframeworks and support the development of targeted therapies. We employed fluorescence insitu hybridization, immunohistochemistry, targeted DNA sequencing, and RNA sequencing toidentify genetic alterations, determine the cell-of-origin, and classify the cell lines intoestablished genetic subtypes.Advanced transcriptomic analyses, including principal component analysis, gene-set enrichmentanalysis and differential gene expression profiling, allowed us to dissect molecular heterogeneityand identify distinct transcriptional programs.In this line, we achieved high concordance in genetic subtype assignment by applying multipleclassification algorithms (2-S, LymphGen and DLBclass), while RNA-seq and DHIT/DZ signatureprofiling further revealed additional molecular complexity. Notably, some DLBCL-NOS casesexhibited high-grade features, and discrepancies in translocation detection suggest that geneexpression signatures may better capture biological aggressiveness than cytogenetic methods.Overall, our study provides a detailed molecular landscape of DLBCL-derived cell lines,demonstrating their relevance as preclinical models for mechanistic investigation and evaluationof therapeutic responses. These findings emphasize the need to integrate genetic andtranscriptomic analyses to refine DLBCL classification and enhance the development ofpersonalized therapeutic strategies.