Deregulated hsa‐miR‐23a‐3p and hsa‐mir‐148a‐3p influence key processes in classic Hodgkin lymphoma (cHL) pathogenesis

Introduction: We previously performed global miRNA expression profiling of cHL vs. non-Hodgkin lymphoma (NHL) cell lines and sorted germinal centre B-cells (GCB). In the group 79 deregulated miRNAs (p < 0.05) we became particularly interested in upregulated miR-23a-3p, a potential oncomiR, predicted to regulate TNFAIP3 downregulated miR-148a-3p, tumor suppressor miRNA, IL15. Therefore, aimed decipher role miR-23a-3p miR-148a-3p pathogenesis. Methods: Expression analyses were with TaqMan Advanced Assays versus two controls. Laser capture microdissection was used for primary HRS collection. 1,000 (miRNA expression) 2 × 200 (DNA methylation) cells collected per case. Bisulfite pyrosequencing assay designed using PyroMark Software sequencing Q24 sequencer. MiRNA-mRNAs interactions validated Dual-Glo Luciferase Assay respective mimics. For overexpression pre-miRNAs cloned into pCDH-CMV-MCS-EF1α-GreenPuro vector transductions triplicate. Cell proliferation measured CCK-8 four replications three independent reactions by GloMax®96 Microplate reader. Results: confirmed 0.001) downregulation 0.014) microdissected (n = 10) GCB 10). Moreover, observed inverse correlation DNA methylation level an adjacent CpG island (r −0.72, p 0.01; n 7, NHL Elevated this detected also 2/6 cases. Next, between their target genes. A reduction 34% 40% 0.01) luciferase signal consequence interaction mimic—TNFAIP3 3’UTR mimic—IL15 3’UTR, respectively. abundance protein reduced after miR-23a (coexpressed miR-27a) L428 0.0018) GCB6-16 0.0083) lines, compared empty transductions. Whereas miR-148a caused 32% decrease KM-H2 line The research funded by: This National Science Center, Poland grants: 2014/14/M/NZ2/00529 M.G. 2019/32/T/NZ5/00441 Etiuda scholarship JP. SH is supported German Research Foundation (DFG, HA6145/3-1). MG, ADz-K NR received funding from European Union’s Horizon 2020 innovation programme under grant agreement No 952304. Keywords: Hodgkin lymphoma, Tumor Biology Heterogeneity conflicts interests pertinent abstract.