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Following our summary on the recommendations for genetic testing in common myeloproliferative neoplasms (MPN) based on the British Society for Haematology (BSH) Good-Practice Paper published by Cross et al.,1 the present summary will focus on the atypical MPN subtypes including chronic eosinophilic leukemia (CEL), myeloid/lymphoid neoplasms with eosinophilia (MLN-eo), chronic neutrophilic leukemia (CNL), MPN-unclassifiable (MPN-U), mastocytosis, and myelodysplastic syndromes (MDS)/MPN overlap.
Briefly, Good-Practice Papers aim to recommend good practice in areas where the evidence is limited but a degree of consensus or uniformity may improve patient care using the Grading of Recommendations Assessment, Development and Evaluation (GRADE) nomenclature.
When a persistent eosinophilia (at least 1.5 × 109/L) with no secondary cause is identified, initial investigation for FIP1L1-PDGFRA on peripheral blood or bone marrow (BM) by fluorescence in situ hybridization (FISH) or nested reverse transcriptase (RT) polymerase chain reaction (PCR) is recommended (Grade 1B).
BM cytogenetics or FISH are recommended to screen for FIP1L1-PDGFRA and other fusion genes, but this must then be confirmed by molecular methods (Grade 1B).
In case of elevated serum tryptase levels without a TK gene fusion, mastocytosis should be considered, and examination of BM histology is essential in this context. Myeloid gene panel and KIT D816V testing should be considered for patients with persistent unexplained eosinophilia who test negative for fusion genes (Grade 2B) (Table 1).
Table 1. Markers of clonality associated with eosinophilia*
Category |
Genes |
Frequency |
---|---|---|
MLN-eo |
FIP1L1-PDGFRA |
5–20% HEUS; >80% MLN-eo |
Other PDGFRA fusions |
Rare |
|
PDGFRB fusions |
<10% MLN-eo |
|
FGFR1 fusions |
<5% MLN-eo |
|
PCM1-JAK2, BCR-JAK2 |
<5% MLN-eo |
|
TK gene fusions in CEL and eosinophilia associated with other MPN or MDS/MPN |
ETV6-ABL1 |
?1–2% HEUS/MPN-eo |
FLT3 fusions |
Rare |
|
Other JAK2 fusions |
Rare |
|
NTRK3 RET, ALK, others |
Very rare |
|
Other variants in CEL and eosinophilia associated with other MPN, MDS/MPN, or SM |
JAK2 exon 13 indels |
1–2% HEUS |
KIT D816V |
3% HEUS |
|
STAT5B N642H |
2% persistent eosinophilia including MPN-eo and MDS/MPN-eo |
|
DNMT3A, TET2, ASXL1, EZH2, SETBP1, CBL, other myeloid genes |
11–22% HES/HEUS |
|
HEUS, hypereosinophilia of undetermined significance; HES, idiopathic hypereosinophilic syndrome; MDS, myelodysplastic syndromes; MPN, myeloproliferative neoplasms; SM, systemic mastocytosis; TK, tyrosine kinase. |
CNL is strongly, but not exclusively, associated with CSF3R mutations which are key diagnostic features, but there is also significant overlap between CNL and MDS/MPN.
In the context of mutated genes, testing for CSF3R variants, preferably as part of a more comprehensive myeloid panel, is recommended for all patients with suspected CNL (Grade 2B).
MPN-U is a rare subtype, mostly consisting of patients who do not meet the diagnostic criteria for a specific MPN subtype, or those with features overlapping with ≥2 subtypes. Most patients test positive for JAK2 V617F, CALR or other myeloid driver mutations.
KIT D816V mutation is present in most (up to 90%) adult patients with systemic mastocytosis (SM); however, variant allele frequency (VAF) is often too low for detection by next-generation sequencing (NGS). Therefore, more sensitive techniques (i.e., RT-PCR or digital PCR) should be used on peripheral blood or BM samples. It is also possible to consider standard mutation analysis on purified mast cell samples.
Sensitive KIT D816V testing is recommended for all patients with a suspected mastocytosis (Grade 1B). If KIT D816V is negative, screening for other KIT mutations should be considered in adult patients and it is recommended in pediatric patients (Grade 1B), as KIT D816V is only seen in 30–50% of children with mastocytosis.
Apart from the KIT driver mutation, additional somatic mutations are noted in 70–90% of advanced SM cases, and most will have an associated hematologic neoplasm (AHN) which is usually of an MDS/MPN subtype.
Certain mutations are associated with adverse prognosis, and as a result, profiling can inform decision making regarding therapy and transplantation.
Myeloid panel analysis is advised in patients with advanced SM who are candidates for allogenic stem cell transplantation (allo-SCT) (Grade 1B).
Myeloid panel analysis should be considered for other SM patients if the behavior of the disease may influence therapy (Grade 2B). Myeloid panel and/or BM cytogenetics should be considered to allow characterisation of the AHN component of SM-AHN (Grade 2B).
Adult MDS/MPN overlap syndromes include chronic myelomonocytic leukemia (CMML), atypical CML BCR-ABL1-negative (aCML), MDS/MPN with ring sideroblasts and thrombocytosis (MDS/MPN-RS-T), and MDS/MPN-U.
It is essential to exclude BCR-ABL1 in all cases, and rearrangements of PDGFRA, PDGFRB, FGFR1, or PCM1-JAK2 in rare cases with an associated eosinophilia (Grade 1B). Cytogenetics should be performed at time of diagnosis as it can demonstrate clonality and provides information on prognosis, and can be useful to rule out any rare TK fusions which can mimic MDS/MPN.
Somatic mutations are reported in >90% of cases across the MDS/MPN overlaps. Therefore, the presence of additional mutations can provide supportive evidence of clonality and inform diagnostic discussions.
A myeloid gene panel and BM cytogenetics or SNP array are recommended for patients diagnosed with MDS/MPN or cases with suspected MDS/MPN but with indeterminate morphology (Grade 1B).
The most frequent genetic mutations seen in MDS/MPN are not specific to these disorders and patients can be further classified investigating genotypic/phenotypic correlations.
Gene panels can provide information on prognosis and are incorporated into prognostic scoring systems across these diseases.
Based on the above information, a targeted sequencing panel is recommended in patients diagnosed with an MDS/MPN overlap disorder, particularly in patients who are candidates for active treatment or transplant. This analysis can also inform prognosis and treatment decisions; therefore, it may be useful even if a patient is on supportive care.
Also, due to a strong correlation between mutations identified in the peripheral blood and BM, especially in CMML, mutational analysis can be a valuable tool in elderly patients or those unfit for BM biopsy.
Mutational analysis represents a valuable component of the diagnostic tool kit to classify patients with atypical MPN subtypes further, and to exclude certain disorders. Investigations using gene panels can also inform decisions around prognosis, and treatment. Genomic and genetic profiling are useful to shed a light on the complex nature of these disorders.
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