PHF6 and DNMT3A mutations are enriched in distinct subgroups of mixed phenotype acute leukemia with T-lineage differentiation by Wenbin Xiao, Maheetha Bharadwaj, Max Levine, Noushin Farnoud, Friederike Pastore, Bartlomiej M. Getta, Anne Hultquist, Christopher Famulare, Juan S. Medina, Minal A. Patel, Qi Gao, Natasha Lewis, Janine Pichardo, Jeeyeon Baik, Brian Shaffer, Sergio Giralt, Raajit Rampal, Sean Devlin, Robert Cimera, Yanming Zhang, Maria E. Arcila, Elli Papaemmanuil, Ross L. Levine, and Mikhail Roshal BloodAdv Volume 2(23):3526-3539 December 11, 2018 © 2018 by The American Society of Hematology

Wenbin Xiao et al. Blood Adv 2018;2:3526-3539 © 2018 by The American Society of Hematology

Diagnosis and subtypes of de novo MPAL Diagnosis and subtypes of de novo MPAL. (A) Inclusion and exclusion criteria of our MPAL cohort. Diagnosis and subtypes of de novo MPAL. (A) Inclusion and exclusion criteria of our MPAL cohort. (B) WHO classification of our MPAL cohort. (C) Overall survival of MPAL. (D) Relapse-free survival of MPAL. Wenbin Xiao et al. Blood Adv 2018;2:3526-3539 © 2018 by The American Society of Hematology

Mutational landscape of MPAL Mutational landscape of MPAL. Asterisks denote genes covered by the FoundationOne Heme panel, but not the RainDance panel, and the frequencies of these mutations in 11 patients were calculated. Mutational landscape of MPAL. Asterisks denote genes covered by the FoundationOne Heme panel, but not the RainDance panel, and the frequencies of these mutations in 11 patients were calculated. Plus signs denote fusions detected by Archer, RNA-seq, and/or FoundationOne Heme panels, and the frequencies of these fusions in 23 patients were calculated. Wenbin Xiao et al. Blood Adv 2018;2:3526-3539 © 2018 by The American Society of Hematology

New molecular subtypes of de novo MPAL New molecular subtypes of de novo MPAL. (A) Graphic demonstration of mutant alleles of PHF6 gene (1 patient had splicing site mutation not illustrated here). New molecular subtypes of de novo MPAL. (A) Graphic demonstration of mutant alleles of PHF6 gene (1 patient had splicing site mutation not illustrated here). (B) New MPAL classification with incorporation of our genetic discoveries. (C-D) Overall survival (C) and relapse-free survival (D) of PHF6 mutated vs DNMT3A unmutated vs others without PHF6 or DNMT3A mutations. Wenbin Xiao et al. Blood Adv 2018;2:3526-3539 © 2018 by The American Society of Hematology

Distribution of somatic mutations in immunophenotypically distinct subpopulations in de novo MPAL. (A) An example of MPAL with T/B/M differentiation (patient 14 in Figure 2; B-blast component is not shown). Distribution of somatic mutations in immunophenotypically distinct subpopulations in de novo MPAL. (A) An example of MPAL with T/B/M differentiation (patient 14 in Figure 2; B-blast component is not shown). (B) An example of MPAL with T/B differentiation (patient 16 in Figure 2). (C-D) An example of T/B/M MPAL by flow cytometry (patient 17). (E-G) VAFs of various mutated genes in sorted blast populations (E, patient 14; F, patient 16; G, patient 17). Lym, sorted natural killer cell population serving as germline reference. Wenbin Xiao et al. Blood Adv 2018;2:3526-3539 © 2018 by The American Society of Hematology