Fig. 1. Identification of the EVL/MIR342 gene locus as a novel candidate regulatory region in hematopoiesis.

a Genomic localization of all therapeutic vector integration sites (IS) close to the EVL/MIR342 gene locus detected in ten Wiskott–Aldrich syndrome gene therapy patients. Histone modifications in human CD34⁺ cells representing promoter (H3K4me3), enhancer (H3K4me1) and active (H3K27ac) chromatin regions. H histone, K Lysine, me methylated, ac acetylated. b Top-ranked miRNAs according to the number of therapeutic vector integration sites (ISs) nearby in efficiently engrafted Wiskott–Aldrich syndrome patients (n = 9). c Percentage of IS, which were sequenced once (light red), more than once (red) and more than twice (dark red) in WAS gene therapy patients during the cause of the study (2097 days post-transplant). d Clonal dynamics of individual gene-corrected cells harboring IS close to the EVL/MIR342 locus, which were repeatedly (>twice, Fig. 1b) detected in WAS patients for up to 2097 days post-transplant. Dashed line represents the median of all repeatedly sequenced IS. e RNA-seq counts for EVL transcripts in defined human hematopoietic cell populations. f Relative expression of Evl (normalized to Tbp) and g miR-342-3p (normalized to Rnu6b) in enriched murine hematopoietic primary populations. Indicated cell populations were isolated from 8- to 10-week-old C57/Bl6 mice using immunophenotypic markers, RNA was isolated and subjected to qRT-PCR. HSC hematopoietic stem cell, MPP multipotent progenitor, CMP common myeloid progenitor, CLP common lymphoid progenitor, GMP granulocyte macrophage progenitor, MEP megakaryocytic erythroide progenitor, Gr granulocyte, Mac macrophage, CD8 T cell, CD4 T cell, NK1.1 natural killer cell, n.d. no data.