Pediatric AML clusters by RNA sequencing from healthy hematopoietic progenitors and overexpresses AML immunotargets such as CD33 and CLEC12A. (A) PCA of 49 samples by RNA sequencing, including 36 AML samples and 13 healthy hematopoietic progenitors. Samples cluster according to disease status, while t(8;21) and KMT2A-mutated samples form clusters within AML samples. One myelodysplastic syndrome–associated AML sample was excluded from this PCA. (B) Volcano plot of all differentially expressed genes with >100 base mean expression according to DESe2 analysis in healthy CD45^dim hematopoietic progenitors and pediatric AML samples. AML immunotargets described by Perna et al⁹  are depicted by red dots. Dotted lines illustrate cutoff values of log2 fold change of 2.5 (2^2,5  ∼5.6) and adjusted P < .01 to define significantly highly enriched genes in sorted AML blasts. (C) Volcano plot detailing differential expression of 35 AML immunotargets described by Perna et al in pediatric AML blasts. (D) Volcano plot of all differentially expressed genes with >100 base mean expression according to DESeq2 analysis in healthy CD45^dimCD34⁺CD38⁻ and pediatric AML samples. AML immunotargets described by Perna et al⁹  are depicted by red dots. Dotted lines illustrate cutoff values of log2 fold change of 2.5 (2^2,5 ∼5.6) and adjusted P < .01 to define significantly highly enriched genes in sorted AML blasts. (E) Volcano plot detailing differential expression of 35 AML immunotargets described by Perna et al in pediatric AML blasts. FC, fold change; HD, healthy donor; mut, mutated.