Polycythemia Vera Evolution to Chronic Myelomocytic Leukemia: The Prognostic Value of Next Generation Sequencing

Abstract

variants: Class 1: relevant in the clinical management of myeloid hemopathies. It has been established as a pathogenic variant in myeloid hemopathies and alters an actionable gene. Class 2: it has been established as a pathogenic variant in solid tumors or non-myeloid hemopathies and alters an actionable gene. Class 3: variant not previously described, affects an actionable gene and in silico predictors or classifies mutations as:• Class 3A: likely pathogenic.• Class 3B: uncertain significance (VUS).• Class 3C: likely benign.† VAF: variant allele frequency.‡ This variant has not been previously described in public databases (either as somatic or germline) and in silico predictors classified it, some as benign and others as pathogenic. In this situation, NGS recommendations suggest to classify these variants as of uncertain significance. Chr = chromosome.

Engraftment characterization of risk-stratified AML in NSGS mice

“… 34-37 In mice engrafted with IR AML samples, the identity of the AML graft was confirmed using targeted next-generation sequencing (NGS), as previously described. 38,39 …”

“…The NGS panel included 32 genes related to myeloid disorders, including those genes known to be mutated at disease presentation. 38,39 Briefly, DNA was extracted from PDX IR AML cells, and a library was prepared using a QIAseq FX DNA Library Kit (QIAGEN). Target regions were then captured and amplified by hybridizing the DNA library with a Myeloid Solution Capture Kit (SOPHiA GENETICS).…”

Divergent leukaemia subclones as cellular models for testing vulnerabilities associated with gains in chromosomes 7, 8 or 18

“…To define the mutation profile of MBU-7 and MBU-8, we sequenced a previously described panel of 32 genes related to myeloid disorders 25 , 26 . Three pathogenic variants of WT1 , TP53 and PTPN11 were observed in both subclones with similar variant allele frequency (Table 3 ).…”

“…Target regions were captured and amplified by hybridizing the DNA library with the Myeloid Solution Capture Kit (Sophia Genetics, Switzerland) and, finally, sequenced using MiSeq platform (Illumina, California). The results were analyzed using Sophia DDM platform 25 , 26 and interpreted using the classification described in Sukhai et al 52 .…”

The Multi-Kinase Inhibitor EC-70124 Is a Promising Candidate for the Treatment of FLT3-ITD-Positive Acute Myeloid Leukemia

“…The captured and amplified DNA was sequenced using NextSeq550 platform (Illumina ® ). The results were analyzed using DataGenomics platform as described elsewhere [37][38][39].…”