{"doi":"10.1016/j.drup.2024.101199","title":"Exploring treatment-driven subclonal evolution of prognostic triple biomarkers: Dual gene fusions and chimeric RNA variants in novel subtypes of acute myeloid leukemia patients with KMT2A rearrangement","abstract":"Chromosomal rearrangements (CR) initiate leukemogenesis in approximately 50% of acute myeloid leukemia (AML) patients; however, limited targeted therapies exist due to a lack of accurate molecular and genetic biomarkers of refractory mechanisms during treatment. Here, we investigated the pathological landscape of treatment resistance and relapse in 16 CR-AML patients by monitoring cytogenetic, RNAseq., and genome-wide changes among newly diagnosed, refractory, and relapsed AML. First, in FISH-diagnosed KMT2A (MLL gene, 11q23)/AFDN (AF6, 6q27)-rearrangement, RNA-sequencing identified an unknown CCDC32 (15q15.1)/CBX3 (7p15.2) gene fusion in both newly diagnosed and relapsed samples, which is previously unknown in KMT2A/AFDN-rearranged AML patients. Second, the unreported CCDC32/CBX3 gene fusion significantly affected the expression of wild-type genes of both CCDC32 (essential for embryonic development) and CBX3 (an oncogene for solid tumors) during the relapse, as demonstrated by Quantitative qPCR analyses. Third, we further confirmed the existence of triple biomarkers - KMT2A/AFDN (AF6, 6q27) rearrangement, the unknown CCDC32 (15q15.1)/CBX3 (7p15.2) gene fusion and chimeric RNA variants (treatment-resistant leukemic blasts harboring distinct breakpoints) in a 21-year-old male patient of rapid relapsed/refractory AML. Most intriguingly, in this work regarding 16 patients, patients 7 and 20 initially showed the KMT2A/AFDN gene fusion; upon relapse, patient 20 did not show this fusion. On the other hand, patient 7 retained the KMT2A/AFDN fusion at diagnosis and during the relapse, only identified by PCR and Sanger's Sequencing, not by cytogenetics. Interestingly, the chimeric CCDC32/CBX3 gene fusion persisted in both individuals over the diagnostic and relapse phases. Most intriguingly, a knock-in experiment confirms the functional validation, providing experimental evidence of the biological impact of the CCDC32/CBX3 fusion on AML pathogenesis and treatment resistance by promoting cell cycle progression, a mechanism through which AML evolves to become treatment-resistant. All these might exhort differential resistance to treatment. Thus, we found that prognostic and predictive triple biomarkers - KRAS mutated, dual fusions (KMT2A/AFDN, CCDC32/CBX3), and chimeric variants - might evolve with a potential oncogenic role of subclonal evolution for poor clinical outcomes. • In AML patients diagnosed with KMT2A/AFDN (AF6, 6q27) rearrangement via FISH, RNA-seq revealed a previously unreported unknown CCDC32 (15q15.1) /CBX3 (7p15.2) gene fusion present, i.e., the dual fusion genes were shown in the same patient. • A knock-in experiment confirms the functional validation, demonstrating that the CCDC32/CBX fusion protein promotes cell cycle progression, a mechanism by which AML evolves to become treatment-resistant. • Our finding of the triple biomarkers (KRAS mutated, the dual oncofusion genes - the unknown -- CCDC32/CBX3 gene fusion (unreported in AML) and CBX3/CCDC32 in cDNA and gDNA), and chimeric RNA variants) in treatment-resistant leukemic blasts with distinct breakpoints might offer treatment-driven subclonal evolution mechanisms for subtypes of relapsed/refractory AML.","journal":"Drug Resistance Updates","year":2025,"id":538382,"datarank":0.0,"base_score":0.0,"endowment":0.0,"self_citation_contribution":0.0,"citation_network_contribution":0.0,"self_endowment_contribution":0.0,"citer_contribution":0.0,"corpus_percentile":null,"corpus_rank":null,"citation_count":3,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9583,"is_data_producer":false,"deposit_databanks":null,"is_oa":true,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":"2025-01-01","fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":525545,"name":"Shengwen Calvin Li","orcid":"0000-0002-9699-9204","position":1,"is_corresponding":false},{"id":1425003,"name":"Jeffrey Xiao","orcid":"0000-0002-6252-3320","position":2,"is_corresponding":false},{"id":499131,"name":"Qian Liu","orcid":"0000-0002-8078-1075","position":3,"is_corresponding":false},{"id":1425372,"name":"Durga Cherukuri","orcid":null,"position":4,"is_corresponding":false},{"id":418652,"name":"Yan Liu","orcid":"0000-0002-8581-2571","position":5,"is_corresponding":false},{"id":1425004,"name":"Saied Mirshahidi","orcid":"0000-0003-1747-8070","position":6,"is_corresponding":false},{"id":1088991,"name":"Jane Xu","orcid":"0000-0003-3619-2676","position":7,"is_corresponding":false},{"id":640876,"name":"Xuelian Chen","orcid":"0000-0003-3850-8363","position":8,"is_corresponding":false},{"id":1425373,"name":"Dadrastoussi Homa","orcid":null,"position":9,"is_corresponding":false},{"id":1425005,"name":"Julian Olea","orcid":"0009-0008-9040-0804","position":10,"is_corresponding":false},{"id":1425006,"name":"Kaijin Wu","orcid":"0009-0005-6604-5775","position":11,"is_corresponding":false},{"id":950363,"name":"Kevin R. Kelly","orcid":"0000-0002-3638-5956","position":12,"is_corresponding":false},{"id":109406,"name":"Fengzhu Sun","orcid":"0000-0002-8552-043X","position":13,"is_corresponding":false},{"id":932451,"name":"Ruihao Huang","orcid":null,"position":14,"is_corresponding":false},{"id":524227,"name":"Xiaoqi Wang","orcid":"0000-0001-8251-9102","position":15,"is_corresponding":false},{"id":932041,"name":"Qin Wen","orcid":"0000-0002-9494-3901","position":16,"is_corresponding":false},{"id":468219,"name":"Xi Zhang","orcid":"0000-0002-4918-0810","position":17,"is_corresponding":false},{"id":856831,"name":"Cristina Maria Ghiuzeli","orcid":null,"position":18,"is_corresponding":false},{"id":1425007,"name":"Esther G Chong","orcid":"0000-0002-0508-9521","position":19,"is_corresponding":false},{"id":249842,"name":"Hisham Abdel‐Azim","orcid":"0000-0003-0880-8701","position":20,"is_corresponding":false},{"id":618581,"name":"Mark E. Reeves","orcid":"0009-0001-1091-0177","position":21,"is_corresponding":false},{"id":1122730,"name":"David J. Baylink","orcid":"0000-0001-9526-7995","position":22,"is_corresponding":false},{"id":932452,"name":"Huynh Cao","orcid":null,"position":23,"is_corresponding":false},{"id":1286163,"name":"Jiang F. Zhong","orcid":"0009-0004-1501-5157","position":24,"is_corresponding":false},{"id":932043,"name":"Yi Xu","orcid":"0000-0003-4215-1993","position":0,"is_corresponding":true}],"reference_count":42,"raw_metadata":null,"created_at":"2026-07-19T02:52:21.389196Z","pmid":"39823827","pmcid":null,"fwci":null,"citation_percentile":null,"influential_citations":0,"oa_status":null,"license":null,"views":0,"total_file_size_bytes":0,"version_count":0,"fair_f":null,"fair_a":null,"fair_i":null,"fair_r":null,"fair_zscore":null,"fair_rationale":null,"fair_model":null,"fair_agent_version":null,"fair_fulltext_source":null,"fair_has_llm":null,"fair_computed_at":null,"clinical_trials":[],"software_tools":[],"db_accessions":[],"linked_datasets":[],"topics":[]}