Identification of AML miRNA signature from patients’ plasma EVs . (A) Workflow for characterization of EVs miRNA content. The RNA content of EVs from blood plasma collected from 12 AML patients and 12 healthy donors were sequenced using Nextseq500 analyzer. (B) Heat map showing the 243 differentially expressed miRNA between the AML patients (orange) and the healthy donors (green) groups. (C) Heat map showing the AML signature composed of 15 miRNAs uniquely upregulated in the AML patients (orange) vs the healthy donors (green). (D) Workflow for validation of miR-1246, a candidate miRNA among the AML signature. Molm-14 cells or PDXs were executed, and miR-1246 levels were measured in the blood plasma EVs by quantitative polymerase chain reaction (qPCR). (E) Correlative analysis at 4 weeks in Molm-14 xenografted mice between human CD45 levels measured by flow cytometer in both the PB and BM (red), and miR-1246 levels measured by qPCR and normalized to U6 control (blue). (F) Survival curve of PDXs (n = 5 per group; PDX1, PDX2, and PDX5). (G) Flow cytometer analysis showing PB chimerism represented by hCD45 levels over time in mice (n = 5 per group) xenografted with PDX1, PDX2, and PDX5. (H) qPCR analysis showing fold change of miR-1246 over time in PDX1 (red), PDX2 (blue), and PDX5 (green) relative to week 2. Data were normalized to U6 control (dCT), and fold change was determined relative to nonengrafted controls (ddCT).