Transcriptomic profiling of double-hit lymphoma preliminarily identifies aberrant ALOX5 captures vulnerability to ferroptosis

Syahru Agung  Setiawan; Mardiah Suci  Hardianti; Chi-Tai  Yeh; Tsu-Yi  Chao; Johan  Kurnianda

doi:10.22146/inajbcs.v57i3.Supplement.24194

Syahru Agung Setiawan Division of Hematology and Medical Oncology, Department of Internal Medicine, Faculty of Medicine, Public Health and Nursing, Gadjah Mada University, Yogyakarta, Indonesia,
Mardiah Suci Hardianti Division of Hematology and Medical Oncology, Department of Internal Medicine, Faculty of Medicine, Public Health and Nursing, Gadjah Mada University, Yogyakarta, Indonesia
Chi-Tai Yeh Department of Medical Research & Education, Taipei Medical University-Shuang Ho Hospital, New Taipei City, Taiwan
Tsu-Yi Chao Department of Hematology & Oncology, Taipei Medical University-Shuang Ho Hospital, New Taipei City, Taiwan
Johan Kurnianda Division of Hematology and Medical Oncology, Department of Internal Medicine, Faculty of Medicine, Public Health and Nursing, Gadjah Mada University, Yogyakarta, Indonesia

DOI: https://doi.org/10.22146/inajbcs.v57i3.Supplement.24194

Keywords: Double-Hit, DHL, DLBCL, RNA sequencing, ALOX5, MYC, Ferroptosis

Abstract

Double-hit lymphoma (DHL) is the most aggressive subset of diffuse large B-cell lymphoma (DLBCL), defined genetically by MYC and BCL2 or BCL6 rearrangements. Despite R-CHOP as the mainstay of treatment, no specific yet effective drug for DHL has been developed, underscoring an unmet medical need. Ferroptosis, an iron-mediated regulated cell death, has emerged as a potential avenue to treat aggressive B-cell lymphomas. However, there are no specific markers to indicate DHL susceptibility to ferroptosis. Using high-throughput transcriptomic profiling, this study elucidated the biomarker of ferroptosis sensitivity in DHL tumors. RNA libraries from biopsy tumors of 22 DLBCL patients diagnosed at TMU-Shuang Ho Hospital, Taiwan, were sequenced on an Illumina NovaSeq platform. A bioinformatics analysis, such as differential gene expression, hierarchical clustering, and functional enrichment, was performed. Published data from 15 DLBCL cell lines from DepMap treated with erastin were re-analyzed to gauge ferroptosis sensitivity. Based on FISH karyotyping, 9 of the 22 patients were classified as DHL, others categorized as Single-Hit Lymphoma (n=1), Triple-Hit Lymphoma (n=1), or DLBCL without rearrangements (n=11). SHL and non-rearrangement cases were grouped as non-DHL (n=12) for comparison against DHL (n=9). Differentially expressed genes highlighted ALOX5 was overexpressed in DHL. Hierarchical clustering revealed distinct DHL-enriched clusters with activated double-hit features, and Myc targets were enriched for ferroptosis-associated genes. DHL-enriched cells showed greater susceptibility to erastin and showed ALOX5 upregulation, suggesting a role in modulating ferroptosis sensitivity. These data reveal ALOX5 as a link between double-hit features and ferroptosis vulnerability. Activation of ALOX5 can drive c-Myc signaling in therapy-resistant cancers and promote lipid peroxidation during ferroptosis. Targeting ALOX5 or using it as a predictive marker could guide ferroptosis-based therapy for DHL in future translational studies.