miRNA landscape in PBL. (A) Volcano plot highlighting DEMs in PBL compared to controls, with significantly upregulated genes (red) and downregulated genes (blue; log₂FC > |1|; Padj < .001). (B) Heatmap displaying unsupervised analysis between PBL (purple) and controls (gray) of 36 DEMs with validated targets based on Affymetrix annotations (log₂FC > |1|; Padj < .001). On top, patients are classified depending on Dx, HIV and EBV infections, and MYC-t. The color of the heat map is represented by the z-score. Three clusters are highlighted on top, PBL-associated clusters (C1 and C2; green) and control-associated cluster (C3; gray). (C) This figure exhibits the proportion of patients in clusters C1 and C2, depending on MYC-t. (D) Enrichment analysis of miRNA targets that were differentially expressed (log₂FC > |1|; Padj < .001) based on hallmark gene sets. Bubble color and size represent –log₁₀(Padj) and gene count, respectively. (E) Venn diagram illustrating the number of overlapping enriched gens between G2/M checkpoint (green), MYC targets (blue), and E2F targets (pink) hallmark data sets. (F) miRNA target network performed with STRING (version 12.0) including the most DEMs (log₂FC > |3|; Padj < .001) and their validated targets according to Affymetrix annotations (represented in circles; log₂FC > |1|; Padj < .05). The colors of the circles represent the hallmark data sets in which each gene is involved, according to the colors of the Venn diagram in panel E. The color of miRNAs represents the downregulation (blue) or upregulation (red). (G) OS of PBL depending on miR-150-5p and E2F3 expression (median normalized expressions, 6.50 and 8.43, respectively). Statistical analysis was performed using Fisher exact test in panel C and the log-rank test in panel G. Dx, diagnosis; N, negative; NA, not available; OS, overall survival; P, positive; TNFA, tumor necrosis factor alpha.