title: High-throughput sequencing detects minimal residual disease in acute T lymphoblastic leukemia.
abstract: High-throughput sequencing (HTS) of lymphoid receptor genes is an emerging technology that can comprehensively assess the diversity of the immune system. Here, we applied HTS to the diagnosis of T-lineage acute lymphoblastic leukemia/lymphoma. Using 43 paired patient samples, we then assessed minimal residual disease (MRD) at day 29 after treatment. The variable regions of TCRB and TCRG were sequenced using an Illumina HiSeq platform after performance of multiplexed polymerase chain reaction, which targeted all potential V-J rearrangement combinations. Pretreatment samples were used to define clonal T cell receptor (TCR) complementarity-determining region 3 (CDR3) sequences, and paired posttreatment samples were evaluated for MRD. Abnormal T lymphoblast identification by multiparametric flow cytometry was concurrently performed for comparison. We found that TCRB and TCRG HTS not only identified clonality at diagnosis in most cases (31 of 43 for TCRB and 27 of 43 for TCRG) but also detected subsequent MRD. As expected, HTS of TCRB and TCRG identified MRD that was not detected by flow cytometry in a subset of cases (25 of 35 HTS compared with 13 of 35, respectively), which highlights the potential of this technology to define lower detection thresholds for MRD that could affect clinical treatment decisions. Thus, next-generation sequencing of lymphoid receptor gene repertoire may improve clinical diagnosis and subsequent MRD monitoring of lymphoproliferative disorders.
authors.affiliation: Department of Laboratory Medicine, University of Washington, Seattle, WA 98109, USA.
authors.forename: David
authors.initials: D
authors.lastname: Wu
authors.forename: Anna
authors.initials: A
authors.lastname: Sherwood
authors.forename: Jonathan R
authors.initials: JR
authors.lastname: Fromm
authors.forename: Stuart S
authors.initials: SS
authors.lastname: Winter
authors.forename: Kimberly P
authors.initials: KP
authors.lastname: Dunsmore
authors.forename: Mignon L
authors.initials: ML
authors.lastname: Loh
authors.forename: Harvey A
authors.initials: HA
authors.lastname: Greisman
authors.forename: Daniel E
authors.initials: DE
authors.lastname: Sabath
authors.forename: Brent L
authors.initials: BL
authors.lastname: Wood
authors.forename: Harlan
authors.initials: H
authors.lastname: Robins
mesh_terms: D001483:Base Sequence; D002986:Clinical Trials as Topic; D002999:Clone Cells; D022801:Complementarity Determining Regions / Q000235:genetics; D005434:Flow Cytometry; D059014:High-Throughput Nucleotide Sequencing / Q000379:methods*; D006801:Humans; D016130:Immunophenotyping; D008969:Molecular Sequence Data; D018365:Neoplasm, Residual; D054218:Precursor T-Cell Lymphoblastic Leukemia-Lymphoma / Q000175:diagnosis* / Q000235:genetics*; D016693:Receptors, Antigen, T-Cell, alpha-beta / Q000235:genetics; D013601:T-Lymphocytes / Q000276:immunology
chemical_list: D022801:Complementarity Determining Regions; D016693:Receptors, Antigen, T-Cell, alpha-beta