{"doi":"10.1002/hon.70093_81","title":"81 | REPRESSION OF miR‐29 VIA MYC LEADS TO INCREASED CD40 SIGNALING IN TRANSFORMED FOLLICULAR LYMPHOMA (FL) AND UNFAVOURABLE PROGNOSIS IN FL","abstract":"Introduction: FL patients are at risk of disease transformation to high-grade lymphoma (tFL). While genetic alterations have been implicated in tFL, the role of microenvironmental interactions and epigenetic regulation by non-coding RNAs remains poorly understood. Results: We performed the first matched profiling of non-coding RNAs (miRNAs) and mRNAs in paired FL and tFL samples (n = 10 pairs). This identified differential expression of 1075 mRNAs and 19 miRNAs, including repression of the miR-29 family in tFL (miR-29a/b/c, Figure A), which we further focused on. We uncovered that MYC activity is uniformly induced in tFL (GSEA, FDR = 0.016) and represses miR-29 by binding to its promotor, and MYC silencing (siRNA) led to miR-29s induction in lymphoma cell lines (p < 0.05). RNAseq in FL-tFL pairs revealed changes in multiple molecular pathways potentially controlled by miR-29s, including CD40 signaling being strongly activated in tFL (GSEA, IPA). CD40 pathway is a major pro-proliferative factor in normal and FL lymph nodes. scRNAseq data reanalysis (Roider et al,2019) revealed that CD40L is amongst the 10 most active ligands in tFL (Figure B). Increased CD40 activation in tFL contrasted with the reduced CD4+ and CD8+ T-cell numbers in tFL niches (IHC in 10 FL-tFL pairs, CIBERSORTx from FL-tFL RNAseq). To directly identify miR-29 targets, we performed RNA profiling in 2 cell lines engineered for miR-29c overexpression, revealing 20 putative miR-29 targets downregulated in both cell lines. This included TRAF4, which has been previously linked to CD40 signaling. miR-29c overexpression leads to ∼50% reduction of TRAF4 levels (Figure C) via miR-29c binding to its 3’UTR. Importantly, cell lines constitutively overexpressing miR-29c or transfected with synthetic miR-29c (1000 nM) were less responsive (↓ pIKKa/b) to recombinant CD40L (Figure C) or HS-5 cells engineered for CD40L expression. Altogether, MYC-mediated miR-29 repression results in increased TRAF4 and CD40 signaling. Importantly, TRAF4 levels were increased in tFL compared to paired FL (n = 11 pairs, Figure D). Ki67 correlated positively with TRAF4 and negatively with miR-29s levels (FL/tFL n = 46), and TRAF4 and MYC were concurrently induced in tFL (R = 0,8, p = 0.004). Lower levels of all miR-29s(a/b/c) were associated with shorter OS and PFS in FL in univariate (n = 185, for miR-29c in Figure E) and multivariate analysis (age, FLIPI, Hgb, LDH, Ann Arbor, B sympt.). Lower miR-29c levels were associated with shorter OS also in a validation FL cohort (n = 92, Figure F) from an R-CHOP arm of a clinical trial (NCT00006721), but not in DLBCL (n = 174). Conclusions: The first whole-genome miRNA profiling in tFL showed that MYC represses miR-29s levels, leading to increased TRAF4 and stronger CD40 signaling propensity (Figure G). This likely represents an adaptive response to reduced CD40L availability from T-cells in the tFL niches. Moreover, low levels of miR-29c can be used as an FFPE-based biomarker of unfavorable FL prognosis. Research funding declaration: Supported by: Czech Science Foundation Grant No.25-15368X; Ministry of Health of the Czech Rep., grant No.NU22-03-00117 and NU23-08-00448; MH CZ-DRO (FNBr, 65269705); MUNI/A/1685/2024; NIH/NCI/NCTN grants U10CA180888, U10CA180819. National Inst. for Cancer Research (programme EXCELES, ID project no.LX22NPO5102)–Funded by the EU–Next Generation EU. Encore Abstract: EHA 2025 Keywords: microenvironment; aggressive B-cell non-Hodgkin lymphoma; indolent non-Hodgkin lymphoma No potential sources of conflict of interest.","journal":"Hematological Oncology","year":2025,"id":568540,"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":0,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9518,"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":1473388,"name":"Katerina Musilova Litzmanova","orcid":null,"position":1,"is_corresponding":false},{"id":1473389,"name":"Filip Kledus","orcid":null,"position":2,"is_corresponding":false},{"id":1221711,"name":"Miroslav Boudný","orcid":"0000-0001-5757-0424","position":3,"is_corresponding":false},{"id":1473390,"name":"Eva Hoferková","orcid":null,"position":4,"is_corresponding":false},{"id":1472809,"name":"Sheena Sharma","orcid":"0000-0002-2316-8725","position":5,"is_corresponding":false},{"id":1472810,"name":"Václav Šeda","orcid":"0000-0003-0234-6470","position":6,"is_corresponding":false},{"id":1472811,"name":"Pedro Faria Zeni","orcid":"0000-0003-0243-5983","position":7,"is_corresponding":false},{"id":1473391,"name":"M. 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