NGS panel enhance precise diagnosis of myeloid neoplasms under WHO-HAEM5 and International Consensus Classification: An observational study

Abstract

This study aimed to assess hematological diseases next-generation sequencing (NGS) panel enhances the diagnosis and classification of myeloid neoplasms (MN) using the 5th edition of the WHO Classification of Hematolymphoid Tumors (WHO-HAEM5) and the International Consensus Classification (ICC) of Myeloid Tumors. A cohort of 112 patients diagnosed with MN according to the revised fourth edition of the WHO classification (WHO-HAEM4R) underwent testing with a 141-gene NGS panel for hematological diseases. Ancillary studies were also conducted, including bone marrow cytomorphology and routine cytogenetics. The cases were then reclassified according to WHO-HAEM5 and ICC to assess the practical impact of these 2 classifications. The mutation detection rates were 93% for acute myeloid leukemia (AML), 89% for myelodysplastic syndrome (MDS), 94% for myeloproliferative neoplasm (MPN), and 100% for myelodysplasia/myeloproliferative neoplasm (MDS/MPN) (WHO-HAEM4R). NGS provided subclassified information for 26 and 29 patients with WHO-HAEM5 and ICC, respectively. In MPN, NGS confirmed diagnoses in 16 cases by detecting JAK2, MPL, or CALR mutations, whereas 13 "triple-negative" MPN cases revealed at least 1 mutation. NGS panel testing for hematological diseases improves the diagnosis and classification of MN. When diagnosed with ICC, NGS produces more classification subtype information than WHO-HAEM5.

Figure 1.

The relationship between WHO-HAEM4R, WHO-HAEM5, and ICC classified MN types. AML-DBD = AML, defined by differentiation, AML-DGA = AML with defining genetic abnormalities, AML-MR = Acute myeloid leukemia, myelodysplasia-related, AML-MRC = AML with myelodysplasia-related changes, AML-MRCA = AML with myelodysplasia‑related cytogenetic abnormalities, AML-MRGM = AML with myelodysplasia‑related gene mutations, AML-RGA = AML with recurrent genetic abnormalities, AML, NOS = AML, not otherwise specified, AML-TP53 = AML with TP53 mutations, CML = chronic myeloid leukemia, CMML = chronic myelomonocytic leukemia, ET = essential thrombocythemia, MDS/AML-MRGM = MDS/AML with myelodysplasia-related gene mutations, MDS-biTP53 = MDS with biallelic TP53 inactivation, MDS-EB = MDS with excess blasts, MDS-IB = MDS with increased blasts, MDS-LB = MDS with low blasts, MDS-MLD = MDS with multilineage dysplasia, MDS,NOS = MDS, not otherwise specified, MDS-SLD = MDS with single lineage dysplasia, MDS/MPN,NOS = MDS/MPN, not otherwise specified, MDS/MPN-RS-T = MDS/MPN with ring sideroblasts and thrombocytosis, MDS/MPN-SF3B1-T = MDS/MPN with thrombocytosis and SF3B1 mutation, MDS/MPN-U = MDS/MPN unclassifiable, MDS-SF3B1, myelodysplastic neoplasm with low blasts and SF3B1 mutation, MDS-TP53 = MDS with TP53 mutation, PMF = primary myelofibrosis, PV = polycythemia vera.

Figure 2.

Organization of mutations into categories of related genes (Orange indicates pathogenic mutations, and blue indicates unknown significance. Dark colors indicate mutations in 2 or more genes, slashes indicate double mutations in the same gene, and crosses indicate multiple mutations in the same gene. Dark green indicates a negative result of NGS, and light green indicates “triple-negative” MPN). Other tumor suppressor genes include RB1, FAT1, SPEN, EGR1, PHF6; Other DNA Methylation gene include: SETD2, DNMT3A, DNMT3B, TPMT; Other tyr kinases gene include: FGFR3, JAK1, JAK3, PDGFRA, ABL1; Ser-thr kinases gene include: ATM, BRAF; Other activated signaling gene includ: ABL2, ANKRD26, ATG2, BCARD11, CBL,CCND1, CCND3, CD79B, CSF1R, CSF3R, CSMD1, ECT2L, ETNK1, IL7R, MYB, NF1, NOTCH1, NOTCH2, NOTCH3, NT5C2, PLCG1, PLCG2, PPM1D, PRDM1, PTPN11, RBBP6, RELN, SAMD9, TNFAIP3, TNFRSF14,TRAF3, TYK2; Other myeloid TF gene include: CREBBP, CUX1, DDX41, ETV6, GATA1, GATA2, IKZF1, MED12, STAT5A, STAT5B, TAL1, TCF3; Other chromatin modifying gene include: ARID1A, ARID5B, ASXL2, BCORL1, DDX41, DIS3, EP300, ID3, KDM6A, KMT2A, KMT2C, KMT2D, PML, SETBP1; Other cohesin gene include: RAD21, SMC1, SMC1A, SMC2, SMC3; Other spliceosome gene include: PRPF8, SF1, SF3A1.