Mutational profiling of isolated myeloid sarcomas and utility of serum 2hg as biomarker of idh1/2 mutations

Access through your institution Buy or subscribe Myeloid sarcoma (MS) is an extramedullary tumor mass consisting of myeloid blasts with or without maturation. MS may precede a characterized

acute myeloid leukemia (AML) or can be present as an isolated tumor. Prognosis of isolated MS is typically poor despite the use of high-dose chemotherapy [1]. As isolated MS are frequently

misdiagnosed for a solid tumor or lymphoma at diagnosis, molecular characterization of MS is not routinely performed, and tailored therapies based on molecular risk stratification in MS

patients remained unused. Like in AML, cytogenetic abnormalities are found in around 50% of MS and mutations in _NPM1_ and _FLT3_ genes are the most frequent molecular lesions reported so

far in MS [1,2,3]. A next-generation sequencing (NGS) assay including 21 genes in a series of 6 MS cases recently detected lesions in a broad spectrum of AML and myelodysplastic syndrome

(MDS)-associated genes critical for epigenetic (_TET2_, _DNMT3A_, _IDH2_, _ASXL1_, _EZH2_), splicing (_SF3B1_), transcription (_RUNX1, WT1_), tumor suppression (_TP53_), or signaling (_KIT_,

_N-RAS_) regulation [4]. To further characterize the mutational landscape of isolated MS, we analyzed gene mutations in cases of isolated MS using an NGS panel of 32 genes recurrently

mutated in myeloid malignancies. Samples were obtained from 14 consecutive patients with histologically proven isolated de novo MS collected in 3 French centers between 2004 and 2016.

Clinical and pathology data are summarized in Table 1. Conventional karyotype on MS sample was normal in the single patient with available data. Genomic DNA (gDNA) was extracted from frozen

MS samples (_n_ = 8) or formalin-fixed paraffin-embedded (FFPE) tissue samples (_n_ = 6) and assessed for mutations in 32 genes (_ASXL1, BCOR, BCORL1, CALR, CBL, CSF3R, DNMT3A, ETV6, EZH2,

FLT3-_TKD_, GATA2, IDH1, IDH2, JAK2, KIT, KRAS, MPL, NPM1, NRAS, PHF6, PTPN11, RIT1, RUNX1, SETBP1, SF3B1, SRSF2, STAG2, TET2, TP53, U2AF1, WT1_, and _ZRSR2_). _FLT3_ internal tandem

duplication (_FLT3_-ITD) mutations were investigated by fluorescent PCR and fragment analysis. _NPM1, ASXL1_ (including c.1934dupG; p.G646WfsX12), _SRSF2_, and _IDH1/2_ variants were

confirmed by Sanger sequencing (cf Supplemental Methods). Overall, 11/14 (79%) patients presented at least 1 mutation in the selected genes. Our results are consistent with the reported high

frequency of _NPM1_ mutation in MS (7/14) [2,3,4]. A _FLT3-_ITD mutation was identified in only 2 out of 10 patients in our cohort, contrasting with the higher frequency in previously

reported cases [2, 4]. Mutations in _IDH1_ and _IDH2_ were predominant molecular features in our MS samples cohort (6/14 patients), including _IDH2_ R140Q mutation in 4/14 patients (29%),

and _IDH1_ R132(H/C) mutation in 2/14 patients (14%). We also detected the common missense mutation _SRSF2 P95(H/L)_ in 3/14 patients (21%). As frequently observed in AML, _NPM1_ mutation

was associated with _IDH1 R132H/C_ or _IDH2 R140Q_ while _SRSF2_ mutation was associated with _IDH2 R140Q_ (cf Supplemental Fig. 1). _KIT_ mutations were found in 2/14 patients (one of which

had positive type A CBFβ-MYH11 fusion transcript on MS sample) in exon 17 (D816H, D820Y) while previously reported in exon 10 [4]. Mutations were also found in _NRAS_ in 2/14 patients

(14%), _PTPN11_ in 2/14 patients (14%), and _TP53, GATA2_, _RUNX1, DNMT3A_ mutations were each found in 1/14 patients (7%). This is a preview of subscription content, access via your

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Article  CAS  Google Scholar  Download references ACKNOWLEDGEMENTS The authors thank Anne Salomon and Elisabeth Chachaty for clinical specimens, Chris Ottolenghi and Lionel Mercier for their

technical assistance in 2HG analysis. This work was partly funded by the Fondation ARC pour la Recherche sur le Cancer. AUTHOR CONTRIBUTIONS CW, JBM, SDB, ClB, CeB, RI, and NB followed the

study participants. CW collected the data and interpreted the results; AR and CQ supervised and performed molecular analysis on MS sample; SB, JD, VP, and AP performed 2HG measures; MTCM

reviewed FDG-PET results; VS and JB performed the cytological and histological analyses; CW wrote the manuscript; AR, AP, VPL, and SDB supervised the project and revised the manuscript. All

authors approved the manuscript. AUTHOR INFORMATION Author notes * These authors contributed equally: Christophe Willekens, Aline Renneville. AUTHORS AND AFFILIATIONS * Département

d’Hématologie, Gustave Roussy, Université Paris-Saclay, F-94805, Villejuif, France Christophe Willekens, Jean-Baptiste Micol & Stéphane De Botton * Inserm U1170, Gustave Roussy,

Université Paris-Saclay, 94805, Villejuif, France Christophe Willekens, Cyril Quivoron, Virginie Penard-Lacronique & Stéphane De Botton * Centre de Biologie-Pathologie, Laboratoire

d’hématologie, Centre Hospitalier Universitaire de Lille, Lille, France Aline Renneville & Claude Preudhomme * Département de Biologie-Pathologie, Gustave Roussy, Université

Paris-Saclay, F-94805, Villejuif, France Sophie Broutin, Véronique Saada, Julia Delahousse, Vianney Poinsignon, Jacques Bosq & Angelo Paci * Hôpital Claude Huriez, Centre Hospitalier

Universitaire Lille, Service des maladies du sang, Lille, France Claire Bories & Céline Berthon * Département d’Hématologie, Hôpital Saint-Louis, Assistance Publique-Hôpitaux de Paris,

Université Paris Diderot, 75475, Paris, France Raphael Itzykson & Nicolas Boissel * Département de médecine nucléaire, Gustave Roussy, Université Paris-Saclay, F-94805, Villejuif, France

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search for this author inPubMed Google Scholar CORRESPONDING AUTHOR Correspondence to Stéphane De Botton. ETHICS DECLARATIONS CONFLICT OF INTEREST SDB has conflict of interest with AGIOS and

CELGENE. No potential conflicts of interest were disclosed by the other authors. ELECTRONIC SUPPLEMENTARY MATERIAL SUPPLEMENTAL FIGURE 1 SUPPLEMENTAL FIGURE 2 FIGURES AND TABLES LEGENDS

SUPPLEMENTAL METHODS RIGHTS AND PERMISSIONS Reprints and permissions ABOUT THIS ARTICLE CITE THIS ARTICLE Willekens, C., Renneville, A., Broutin, S. _et al._ Mutational profiling of isolated

myeloid sarcomas and utility of serum 2HG as biomarker of _IDH1/2_ mutations. _Leukemia_ 32, 2008–2081 (2018). https://doi.org/10.1038/s41375-018-0056-6 Download citation * Received: 30

November 2017 * Revised: 08 January 2018 * Accepted: 12 January 2018 * Published: 26 February 2018 * Issue Date: September 2018 * DOI: https://doi.org/10.1038/s41375-018-0056-6 SHARE THIS

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