title: Next-generation sequencing of peripheral B-lineage cells pinpoints the circulating clonotypic cell pool in multiple myeloma.
abstract: The identity of the proliferative compartment of myeloma progenitor cells remains a matter of debate. Polymerase chain reaction-based studies suggested pre-switch "clonotypic" B cells sharing the immunoglobulin (Ig) rearrangement of the malignant plasma cell (M-PC), to circulate in the blood and possess stem cell-like properties. Here, we disprove this hypothesis. We screened peripheral blood IgM, IgG, and IgA repertoires of myeloma patients for the clonotypic rearrangement by next-generation sequencing. None of 12 cases showed pre-switch clonotypic transcripts. In the post-switch IgG/IgA repertoires, however, the clonotypic rearrangement was detected at high frequency in 6 of 8 patients with active disease, whereas it was undetectable after treatment, correlating with flow cytometric presence or absence of circulating M-PCs. Minor subclones with alternative post-switch isotypes suggested ongoing switch events and clonal evolution at the M-PC level. Our findings consistently show an absence of pre-switch clonotypic B cells, while M-PCs circulate in the peripheral blood and may contribute to spreading of the disease.
authors.affiliation: University Medical Center Hamburg-Eppendorf, Hubertus Wald Tumorzentrum, University Cancer Center Hamburg, Department of Oncology, Hematology, Bone Marrow Transplantation with Section Pneumology, Hamburg, Germany;
authors.forename: Benjamin
authors.initials: B
authors.lastname: Thiele
authors.affiliation: University Medical Center Hamburg-Eppendorf, Hubertus Wald Tumorzentrum, University Cancer Center Hamburg, Department of Oncology, Hematology, Bone Marrow Transplantation with Section Pneumology, Hamburg, Germany;
authors.forename: Marie
authors.initials: M
authors.lastname: Kloster
authors.affiliation: University Medical Center Hamburg-Eppendorf, Bioinformatics Service Facility, Hamburg, Germany; and.
authors.forename: Malik
authors.initials: M
authors.lastname: Alawi
authors.affiliation: Heinrich-Pette-Institute, Leibniz-Institute for Experimental Virology, Hamburg, Germany.
authors.forename: Daniela
authors.initials: D
authors.lastname: Indenbirken
authors.affiliation: University Medical Center Hamburg-Eppendorf, Hubertus Wald Tumorzentrum, University Cancer Center Hamburg, Department of Oncology, Hematology, Bone Marrow Transplantation with Section Pneumology, Hamburg, Germany;
authors.forename: Martin
authors.initials: M
authors.lastname: Trepel
authors.affiliation: Heinrich-Pette-Institute, Leibniz-Institute for Experimental Virology, Hamburg, Germany.
authors.forename: Adam
authors.initials: A
authors.lastname: Grundhoff
authors.affiliation: University Medical Center Hamburg-Eppendorf, Hubertus Wald Tumorzentrum, University Cancer Center Hamburg, Department of Oncology, Hematology, Bone Marrow Transplantation with Section Pneumology, Hamburg, Germany;
authors.forename: Mascha
authors.initials: M
authors.lastname: Binder
mesh_terms: D001402:B-Lymphocytes / Q000378:metabolism*; D001773:Blood Cells / Q000378:metabolism; D019070:Cell Lineage / Q000235:genetics*; D002999:Clone Cells; D015331:Cohort Studies; D015322:Gene Rearrangement, B-Lymphocyte / Q000235:genetics; D059014:High-Throughput Nucleotide Sequencing; D006801:Humans; D017578:Immunoglobulin Class Switching; D007136:Immunoglobulins / Q000235:genetics; D009101:Multiple Myeloma / Q000235:genetics* / Q000473:pathology*; D010641:Phenotype; D010950:Plasma Cells / Q000378:metabolism / Q000473:pathology
chemical_list: D007136:Immunoglobulins