A predictive serum miRNA signature impacts diffuse large B‐cell lymphoma cell viability via inhibition of EGLN1 and TXNRD1 regulators of ferroptosis

• Published in: British journal of haematology Vol. 207; no. 3; pp. 802 - 812
• Main Authors: Regazzo, Giulia, Vari, Giulia, Marchesi, Francesco, Díaz Méndez, Ana Belén, Di Giuliani, Marta, Sacconi, Andrea, Palombi, Francesca, Lulli, Valentina, Goeman, Frauke, Novello, Mariangela, Tomassi, Martina, Papa, Elena, Bertoni, Francesco, Hohaus, Stefan, Mengarelli, Andrea, Rizzo, Maria Giulia
• Format: Journal Article
• Language: English
• Published: England Blackwell Publishing Ltd 01.09.2025
• Subjects: Adult; Aged; Antineoplastic Combined Chemotherapy Protocols - therapeutic use; Biomarkers; Biomarkers, Tumor - blood; Cell Line, Tumor; Cell proliferation; Cell Survival - genetics; Cell viability; circulating microRNAs; Cyclophosphamide - therapeutic use; diffuse large B‐cell lymphoma; Doxorubicin - therapeutic use; Female; Ferroptosis; Ferroptosis - genetics; ferroptosis regulators; Gene Expression Regulation, Neoplastic; Germinal centers; Homeostasis; Humans; Lymphoma; Lymphoma, Large B-Cell, Diffuse - blood; Lymphoma, Large B-Cell, Diffuse - drug therapy; Lymphoma, Large B-Cell, Diffuse - genetics; Lymphoma, Large B-Cell, Diffuse - pathology; Male; Medical prognosis; MicroRNAs; MicroRNAs - blood; MicroRNAs - genetics; Middle Aged; miRNA; Neoplasm Proteins; predictive biomarkers; Prednisone - therapeutic use; Prognosis; Rituximab; RNA, Neoplasm - blood; Therapeutic applications; Thioredoxin Reductase 1 - antagonists & inhibitors; Thioredoxin Reductase 1 - genetics; Thioredoxin Reductase 1 - metabolism; Vincristine - therapeutic use; ferroptosis regulators; predictive biomarkers; circulating microRNAs; diffuse large B‐cell lymphoma
• ISSN: 0007-1048; 1365-2141; 1365-2141
• DOI: 10.1111/bjh.70017

Summary Diffuse large B‐cell lymphoma (DLBCL) is a heterogeneous disorder. Prognostic factors include genomic alterations and cell‐of‐origin (COO) subtypes, even though they cannot fully predict treatment response. MicroRNAs (miRNAs) deregulated in patient tumours and blood are promising non‐invasive biomarkers. Several circulating miRNAs were found to be correlated with progression‐free survival (PFS), independently of other prognosticators. However, miRNA signatures, rather than individual miRNAs, represent more reliable biomarkers and a better mirror of the disease. In this study, we identified circulating miRNAs differentially expressed between R‐CHOP refractory and responding subjects by small‐RNA sequencing on serum from 33 DLBCL patients. Among the identified miRNAs, the combined expression of three of them improved the predictive performance and was correlated with PFS. Two out of three miRNAs, miR‐421 and miR‐324‐5p, were also differentially expressed in tumour tissues based on treatment response. Overexpressing these miRNAs reduced cell proliferation, viability and resistance to R‐CHOP in the germinal centre B‐like COO subtype. EGLN1 and TXNRD1, regulators of oxygen metabolism and redox homeostasis, were identified as miRNA targets and the silencing or inhibition of these genes impaired cell viability and induced ferroptosis. These results support the application of a two‐miRNA signature and its targets for novel combined therapeutic interventions in DLBCL. Diffuse large B‐cell lymphoma is a heterogeneous and aggressive pathology. MicroRNAs, deregulated in patient tumours and blood, are promising non‐invasive predictive biomarkers for this pathology. We identified the circulating miR‐200c/421/324 signature both predictive of response to R‐CHOP and correlated with PFS. MiR‐421 and miR‐324‐5p were also altered in tumour tissues based on treatment response. Their overexpression reduced cell proliferation, viability and resistance to R‐CHOP in the germinal centre B‐like cell‐of‐origin subtype. The miR‐421/324 signature elicited its biological effects by induction of ferroptosis through the modulation of EGLN1 and TXNRD1 target genes, involved in the regulation of the redox homeostasis.