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Multiple myeloma gammopathies

Circulating tumor cells for comprehensive and multiregional non-invasive genetic characterization of multiple myeloma


Multiple myeloma (MM) patients undergo repetitive bone marrow (BM) aspirates for genetic characterization. Circulating tumor cells (CTCs) are detectable in peripheral blood (PB) of virtually all MM cases and are prognostic, but their applicability for noninvasive screening has been poorly investigated. Here, we used next-generation flow (NGF) cytometry to isolate matched CTCs and BM tumor cells from 53 patients and compared their genetic profile. In eight cases, tumor cells from extramedullary (EM) plasmacytomas were also sorted and whole-exome sequencing was performed in the three spatially distributed tumor samples. CTCs were detectable by NGF in the PB of all patients with MM. Based on the cancer cell fraction of clonal and subclonal mutations, we found that ~22% of CTCs egressed from a BM (or EM) site distant from the matched BM aspirate. Concordance between BM tumor cells and CTCs was high for chromosome arm-level copy number alterations (≥95%) though not for translocations (39%). All high-risk genetic abnormalities except one t(4;14) were detected in CTCs whenever present in BM tumor cells. Noteworthy, ≥82% mutations present in BM and EM clones were detectable in CTCs. Altogether, these results support CTCs for noninvasive risk-stratification of MM patients based on their numbers and genetic profile.

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Fig. 1: Spatial mutational heterogeneity in patients with extramedullary disease.
Fig. 2: CTCs harbor most mutations present in both medullary and extramedullary disease.
Fig. 3: Cancer cell fraction of mutations in matched CTCs and BM tumor cells.
Fig. 4: Concordance between mutations, CNA, and translocations found in CTCs vs BM tumor cells.
Fig. 5: CNA in matched BM tumor cells and CTCs using microarrays.

Data availability

WES and arrays data that support the findings of this study have been deposited in the European Genome-phenome Archive (ENA) with the following accession codes: EGAS00001004288 and EGAS00001004314. Data will be available immediately following publication, no end date. All other data and processing code are available from the corresponding author upon reasonable request.


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We would like to thank to the patients and their families who participated in this study.


This study was supported by the Centro de Investigación Biomédica en Red—Área de Oncología—del Instituto de Salud Carlos III (CIBERONC; CB16/12/00236, CB16/12/00369, CB16/12/00489, and CB16/12/00400); by Cancer Research UK [C355/A26819] and FC AECC and AIRC under the Accelerator Award Program; by the Instituto de Salud Carlos III, FCAECC and co-financed by FEDER (ERANET-TRANSCAN-2 iMMunocell AC17/00101); the Spanish Ministry of Science and Innovation and co-financed by FSE (Torres Quevedo fellowship, PTQ-16-08623); the Black Swan Research Initiative of the International Myeloma Foundation; European Research Council (ERC) under the European Commission’s H2020 Framework Programme (MYELOMANEXT, 680200); the Qatar National Research Fund (QNRF) Award No. 7-916-3-237; the AACR-Millennium Fellowship in Multiple Myeloma Research (15-40-38-PAIV); the Leukemia Research Foundation; and the Multiple Myeloma Research Foundation (MMRF) under the 2019 Research Fellowship Award.

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Authors and Affiliations




GB, CLO, JFSM, and BP conceived the idea and designed the study protocol; NP, M-TC, M-JC, XA, JFM, LSF, PRO, RR, JML, PM, LP, RDO, APM, HEO, FP, M-VM, LR, JB, J-JL, AO, and JFSM provided study material and patients; LB, NP, M-TC, and BP analyzed flow cytometry data; DA and IR performed cell sorting; JJG, GB, LB, DAP, IG, and SR extracted and processed the samples; JJG, GB, RV-M, and M-GA did/supervised the bioinformatics processing and JJG, GB, RV-M, and BP analyzed and interpreted data; JJG, GB, RV-M, and BP wrote the manuscript and all authors reviewed and approved the manuscript.

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Correspondence to Bruno Paiva.

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Conflict of interest

M-JC reports membership on board of directors or advisory committees with Janssen, Celgene, and Novartis. PRO reports membership on board of directors or advisory committees with Janssen, Celgene, Takeda, Amgen, Abbvie, Sanofi, BMS, and Kite Pharma. M-VM reports membership on board of directors or advisory committees with Janssen, Celgene, Takeda, Amgen, Abbvie, GSK, Pharmamar, Mundipharma-EDO, Adaptive, and Seattle-Genetics. LR reports honoraria from Janssen, Celgene, Amgen, and Takeda. JB reports honoraria from Celgene, Amgen, and Janssen. J-JL reports honoraria from and membership on board of directors or advisory committees with Takeda, Amgen, Celgene, and Janssen. JFSM reports consultancy for Bristol-Myers Squibb, Celgene, Novartis, Takeda, Amgen, MSD, Janssen, and Sanofi and membership on board of directors or advisory committees with Takeda. BP honoraria for lectures from and membership on advisory boards with Amgen, Bristol-Myers Squibb, Celgene, Janssen, Merck, Novartis, Roche, and Sanofi; unrestricted grants from Celgene, EngMab, Sanofi, and Takeda; and consultancy for Celgene, Janssen, and Sanofi.

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Garcés, JJ., Bretones, G., Burgos, L. et al. Circulating tumor cells for comprehensive and multiregional non-invasive genetic characterization of multiple myeloma. Leukemia 34, 3007–3018 (2020).

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