title: Clonal architecture of secondary acute myeloid leukemia defined by single-cell sequencing.
abstract: Next-generation sequencing has been used to infer the clonality of heterogeneous tumor samples. These analyses yield specific predictions-the population frequency of individual clones, their genetic composition, and their evolutionary relationships-which we set out to test by sequencing individual cells from three subjects diagnosed with secondary acute myeloid leukemia, each of whom had been previously characterized by whole genome sequencing of unfractionated tumor samples. Single-cell mutation profiling strongly supported the clonal architecture implied by the analysis of bulk material. In addition, it resolved the clonal assignment of single nucleotide variants that had been initially ambiguous and identified areas of previously unappreciated complexity. Accordingly, we find that many of the key assumptions underlying the analysis of tumor clonality by deep sequencing of unfractionated material are valid. Furthermore, we illustrate a single-cell sequencing strategy for interrogating the clonal relationships among known variants that is cost-effective, scalable, and adaptable to the analysis of both hematopoietic and solid tumors, or any heterogeneous population of cells.
authors.affiliation: Center for Genome Sciences and Systems Biology, Washington University, St. Louis, Missouri, United States of America.
authors.forename: Andrew E O
authors.initials: AE
authors.lastname: Hughes
authors.affiliation: The Genome Institute, Washington University, St. Louis, Missouri, United States of America; Department of Genetics, Washington University, St. Louis, Missouri, United States of America.
authors.forename: Vincent
authors.initials: V
authors.lastname: Magrini
authors.affiliation: The Genome Institute, Washington University, St. Louis, Missouri, United States of America.
authors.forename: Ryan
authors.initials: R
authors.lastname: Demeter
authors.affiliation: The Genome Institute, Washington University, St. Louis, Missouri, United States of America.
authors.forename: Christopher A
authors.initials: CA
authors.lastname: Miller
authors.affiliation: The Genome Institute, Washington University, St. Louis, Missouri, United States of America.
authors.forename: Robert
authors.initials: R
authors.lastname: Fulton
authors.affiliation: The Genome Institute, Washington University, St. Louis, Missouri, United States of America.
authors.forename: Lucinda L
authors.initials: LL
authors.lastname: Fulton
authors.affiliation: Department of Internal Medicine, Division of Oncology, Washington University, St. Louis, Missouri, United States of America.
authors.forename: William C
authors.initials: WC
authors.lastname: Eades
authors.affiliation: Department of Internal Medicine, Division of Oncology, Washington University, St. Louis, Missouri, United States of America.
authors.forename: Kevin
authors.initials: K
authors.lastname: Elliott
authors.affiliation: Department of Internal Medicine, Division of Oncology, Washington University, St. Louis, Missouri, United States of America.
authors.forename: Sharon
authors.initials: S
authors.lastname: Heath
authors.affiliation: Department of Internal Medicine, Division of Oncology, Washington University, St. Louis, Missouri, United States of America; Siteman Cancer Center, Washington University, St. Louis, Missouri, United States of America.
authors.forename: Peter
authors.initials: P
authors.lastname: Westervelt
authors.affiliation: The Genome Institute, Washington University, St. Louis, Missouri, United States of America; Department of Internal Medicine, Division of Oncology, Washington University, St. Louis, Missouri, United States of America.
authors.forename: Li
authors.initials: L
authors.lastname: Ding
authors.affiliation: Department of Genetics, Washington University, St. Louis, Missouri, United States of America; Department of Pathology and Immunology, Washington University, St. Louis, Missouri, United States of America.
authors.forename: Donald F
authors.initials: DF
authors.lastname: Conrad
authors.affiliation: The Genome Institute, Washington University, St. Louis, Missouri, United States of America; Department of Internal Medicine, Division of Oncology, Washington University, St. Louis, Missouri, United States of America.
authors.forename: Brian S
authors.initials: BS
authors.lastname: White
authors.affiliation: Department of Internal Medicine, Division of Oncology, Washington University, St. Louis, Missouri, United States of America; Siteman Cancer Center, Washington University, St. Louis, Missouri, United States of America.
authors.forename: Jin
authors.initials: J
authors.lastname: Shao
authors.affiliation: Department of Internal Medicine, Division of Oncology, Washington University, St. Louis, Missouri, United States of America; Siteman Cancer Center, Washington University, St. Louis, Missouri, United States of America.
authors.forename: Daniel C
authors.initials: DC
authors.lastname: Link
authors.affiliation: Department of Internal Medicine, Division of Oncology, Washington University, St. Louis, Missouri, United States of America; Siteman Cancer Center, Washington University, St. Louis, Missouri, United States of America.
authors.forename: John F
authors.initials: JF
authors.lastname: DiPersio
authors.affiliation: The Genome Institute, Washington University, St. Louis, Missouri, United States of America; Department of Genetics, Washington University, St. Louis, Missouri, United States of America.
authors.forename: Elaine R
authors.initials: ER
authors.lastname: Mardis
authors.affiliation: The Genome Institute, Washington University, St. Louis, Missouri, United States of America; Department of Genetics, Washington University, St. Louis, Missouri, United States of America; Siteman Cancer Center, Washington University, St. Louis, Missouri, United States of America.
authors.forename: Richard K
authors.initials: RK
authors.lastname: Wilson
authors.affiliation: The Genome Institute, Washington University, St. Louis, Missouri, United States of America; Department of Genetics, Washington University, St. Louis, Missouri, United States of America; Department of Internal Medicine, Division of Oncology, Washington University, St. Louis, Missouri, United States of America; Siteman Cancer Center, Washington University, St. Louis, Missouri, United States of America.
authors.forename: Timothy J
authors.initials: TJ
authors.lastname: Ley
authors.affiliation: Department of Genetics, Washington University, St. Louis, Missouri, United States of America; Department of Internal Medicine, Division of Oncology, Washington University, St. Louis, Missouri, United States of America; Siteman Cancer Center, Washington University, St. Louis, Missouri, United States of America.
authors.forename: Matthew J
authors.initials: MJ
authors.lastname: Walter
authors.affiliation: Department of Internal Medicine, Division of Oncology, Washington University, St. Louis, Missouri, United States of America; Siteman Cancer Center, Washington University, St. Louis, Missouri, United States of America; Department of Pathology and Immunology, Washington University, St. Louis, Missouri, United States of America.
authors.forename: Timothy A
authors.initials: TA
authors.lastname: Graubert
mesh_terms: D000328:Adult; D000368:Aged; D045744:Cell Line, Tumor; D060965:Clonal Evolution / Q000235:genetics*; D002999:Clone Cells; D005260:Female; D005787:Gene Frequency; D059014:High-Throughput Nucleotide Sequencing; D006801:Humans; D015470:Leukemia, Myeloid, Acute / Q000235:genetics* / Q000473:pathology; D008297:Male; D008875:Middle Aged; D009154:Mutation; D009190:Myelodysplastic Syndromes / Q000235:genetics / Q000473:pathology; D009364:Neoplasm Recurrence, Local / Q000235:genetics / Q000473:pathology; D020641:Polymorphism, Single Nucleotide; D059010:Single-Cell Analysis