Targeted NGS on Sequential Bone Marrow Biopsies Aids in the Evaluation of Cytopenias and Monocytosis and Documents Clonal Evolution-a Proof of Principle Study

Overview

Journal Virchows Arch

Specialties Cell Biology
Molecular Biology
Pathology

Date 2023 Aug 23

PMID 37610626

Authors

Dominik Nann

Achim Rau

Lejla Mahmutovic

Julia Steinhilber

Vanessa Meca

Birgit Federmann

Wichard Vogel

Irina Bonzheim

Leticia Quintanilla-Martinez

Falko Fend

Affiliations

Soon will be listed here.

Abstract

Differential diagnosis of clonal versus reactive cytopenia and monocytosis, respectively, frequently presents a diagnostic challenge. With the two recent classifications of myeloid disorders, mutational analysis has gained importance as a diagnostic tool. However, reports on its utility on trephine bone marrow biopsies (BMB) are sparse. The aim of our proof of principle study was to determine the suitability of targeted sequencing for the longitudinal evaluation of cytopenia and monocytosis and demonstration of clonal evolution on sequential BMB. Seventy-seven EDTA-decalcified BMB of 33 patients with peripheral cytopenia and/or monocytosis, including at least one follow-up biopsy/patient, were included. Initial morphological diagnoses were idiopathic cytopenia of undetermined significance (ICUS, 8 cases), MDS (without blast increase, 7 cases), MDS with increased blasts/excess blasts (MDS-IB/EB) (11 cases), and CMML (7 cases). Thirty-one genes relevant for myeloid disorders were examined using two custom AmpliSeq NGS panels. Mutations were found in the initial BMB of 5/8 cases of ICUS, thus changing the diagnosis to clonal cytopenia of unknown significance (CCUS), 5/7 MDS, 10/11 MDS-IB/EB, and 7/7 CMML. Clonal evolution was observed in 14/33 (42%) cases, mostly associated with disease progression. None of the wild-type patients acquired mutations during follow-up. NGS-based mutation profiling is a robust diagnostic tool for BMB and provides valuable additional information, especially for cases with no/minimal dysplasia, and for better risk stratification of MDS. Tracking variant allele frequency and appearance of mutations over time allows for observing clonal evolution or relapse.

References

Bejar R . Clinical and genetic predictors of prognosis in myelodysplastic syndromes. Haematologica. 2014; 99(6):956-64. PMC: 4040892. DOI: 10.3324/haematol.2013.085217. View

Khoury J, Solary E, Abla O, Akkari Y, Alaggio R, Apperley J . The 5th edition of the World Health Organization Classification of Haematolymphoid Tumours: Myeloid and Histiocytic/Dendritic Neoplasms. Leukemia. 2022; 36(7):1703-1719. PMC: 9252913. DOI: 10.1038/s41375-022-01613-1. View

Huber S, Haferlach T, Muller H, Meggendorfer M, Hutter S, Hoermann G . MDS subclassification-do we still have to count blasts?. Leukemia. 2023; 37(4):942-945. PMC: 10079547. DOI: 10.1038/s41375-023-01855-7. View

Sperling A, Gibson C, Ebert B . The genetics of myelodysplastic syndrome: from clonal haematopoiesis to secondary leukaemia. Nat Rev Cancer. 2016; 17(1):5-19. PMC: 5470392. DOI: 10.1038/nrc.2016.112. View

Kwok B, Hall J, Witte J, Xu Y, Reddy P, Lin K . MDS-associated somatic mutations and clonal hematopoiesis are common in idiopathic cytopenias of undetermined significance. Blood. 2015; 126(21):2355-61. PMC: 4653764. DOI: 10.1182/blood-2015-08-667063. View

Bartels S, Schipper E, Kreipe H, Lehmann U . Comprehensive Molecular Profiling of Archival Bone Marrow Trephines Using a Commercially Available Leukemia Panel and Semiconductor-Based Targeted Resequencing. PLoS One. 2015; 10(7):e0133930. PMC: 4519100. DOI: 10.1371/journal.pone.0133930. View

Walter M, Shen D, Shao J, Ding L, White B, Kandoth C . Clonal diversity of recurrently mutated genes in myelodysplastic syndromes. Leukemia. 2013; 27(6):1275-82. PMC: 3736571. DOI: 10.1038/leu.2013.58. View

Haase D, Germing U, Schanz J, Pfeilstocker M, Nosslinger T, Hildebrandt B . New insights into the prognostic impact of the karyotype in MDS and correlation with subtypes: evidence from a core dataset of 2124 patients. Blood. 2007; 110(13):4385-95. DOI: 10.1182/blood-2007-03-082404. View

Hosono N, Chi S, Yamauchi T, Fukushima K, Shibayama H, Katagiri S . Clinical utility of genomic profiling of AML using paraffin-embedded bone marrow clots: HM-SCREEN-Japan 01. Cancer Sci. 2023; 114(5):2098-2108. PMC: 10154825. DOI: 10.1111/cas.15746. View

10.

Germing U, Strupp C, Kundgen A, Bowen D, Aul C, Haas R . No increase in age-specific incidence of myelodysplastic syndromes. Haematologica. 2004; 89(8):905-10. View

11.

Patnaik M, Tefferi A . Chronic myelomonocytic leukemia: 2018 update on diagnosis, risk stratification and management. Am J Hematol. 2018; 93(6):824-840. PMC: 5995129. DOI: 10.1002/ajh.25104. View

12.

Patnaik M, Wassie E, Padron E, Onida F, Itzykson R, Lasho T . Chronic myelomonocytic leukemia in younger patients: molecular and cytogenetic predictors of survival and treatment outcome. Blood Cancer J. 2015; 5:e270. PMC: 5404217. DOI: 10.1038/bcj.2014.90. View

13.

Hasemeier B, Geffers R, Bartels S, Schlegelberger B, Kreipe H, Lehmann U . Archival bone marrow trephines are suitable for high-throughput mutation analysis using next generation sequencing technology. Haematologica. 2013; 98(9):e115-6. PMC: 3762114. DOI: 10.3324/haematol.2013.091652. View

14.

Cogle C, Craig B, Rollison D, List A . Incidence of the myelodysplastic syndromes using a novel claims-based algorithm: high number of uncaptured cases by cancer registries. Blood. 2011; 117(26):7121-5. PMC: 3143554. DOI: 10.1182/blood-2011-02-337964. View

15.

Jaiswal S, Ebert B . Clonal hematopoiesis in human aging and disease. Science. 2019; 366(6465). PMC: 8050831. DOI: 10.1126/science.aan4673. View

16.

Itzykson R, Kosmider O, Renneville A, Gelsi-Boyer V, Meggendorfer M, Morabito M . Prognostic score including gene mutations in chronic myelomonocytic leukemia. J Clin Oncol. 2013; 31(19):2428-36. DOI: 10.1200/JCO.2012.47.3314. View

17.

Mughal T, Cross N, Padron E, Tiu R, Savona M, Malcovati L . An International MDS/MPN Working Group's perspective and recommendations on molecular pathogenesis, diagnosis and clinical characterization of myelodysplastic/myeloproliferative neoplasms. Haematologica. 2015; 100(9):1117-30. PMC: 4800699. DOI: 10.3324/haematol.2014.114660. View

18.

Bartels S, Hasemeier B, Vogtmann J, Schipper E, Busche G, Schlue J . Feasibility of Combined Detection of Gene Mutations and Fusion Transcripts in Bone Marrow Trephines from Leukemic Neoplasms. J Mol Diagn. 2020; 22(4):591-598. DOI: 10.1016/j.jmoldx.2020.01.004. View

19.

Bernard V, Gebauer N, Dinh T, Stegemann J, Feller A, Merz H . Applicability of next-generation sequencing to decalcified formalin-fixed and paraffin-embedded chronic myelomonocytic leukaemia samples. Int J Clin Exp Pathol. 2014; 7(4):1667-76. PMC: 4014247. View

20.

DeZern A, Malcovati L, Ebert B . CHIP, CCUS, and Other Acronyms: Definition, Implications, and Impact on Practice. Am Soc Clin Oncol Educ Book. 2019; 39:400-410. DOI: 10.1200/EDBK_239083. View