title: Single cell genotyping of exome sequencing-identified mutations to characterize the clonal composition and evolution of inv(16) AML in a CBL mutated clonal hematopoiesis.
abstract: We recently described the development of an inv(16) acute myeloid leukemia (AML) in a CBL mutated clonal hematopoiesis. Here, we further characterized the clonal composition and evolution of the AML based on the genetic information from the bulk specimen and analyses of individual bone marrow cells for mutations in CAND1, PTPRT, and DOCK6. To control for allele dropout, heterozygous polymorphisms located close to the respective mutation loci were assessed in parallel. The clonal composition concluded from exome sequencing suggested a proliferation advantage associated with the acquisition of mutations in CAND1, PTPRT, and DOCK6. Out of 102 single cell sequencing reactions on these mutations and the respective polymorphisms, analyses yielded conclusive results for at least 2 mutation sites in 12 cells. The single cell genotyping not only confirmed the co-occurrence of the PTPRT, CAND1 and DOCK6 mutations in the same AML clone but also revealed a clonal hierarchy, as the PTPRT mutation was likely acquired after the CAND1 and DOCK6 mutations. This insight had not been possible based solely on the exome sequencing data and suggests that the mutation in PTPRT, which encodes a STAT3-inhibiting protein tyrosine phosphatase, contributed to the AML development at a later stage by enhancing proliferation.
authors.affiliation: Department of Internal Medicine I, University Freiburg-Medical Center, Faculty of Medicine, Freiburg, Germany.
authors.forename: Christoph
authors.initials: C
authors.lastname: Niemöller
authors.affiliation: Department of Internal Medicine I, University Freiburg-Medical Center, Faculty of Medicine, Freiburg, Germany.
authors.forename: Nathalie
authors.initials: N
authors.lastname: Renz
authors.affiliation: Department of Internal Medicine I, University Freiburg-Medical Center, Faculty of Medicine, Freiburg, Germany.
authors.forename: Sabine
authors.initials: S
authors.lastname: Bleul
authors.affiliation: Department of Internal Medicine I, University Freiburg-Medical Center, Faculty of Medicine, Freiburg, Germany.
authors.forename: Nadja
authors.initials: N
authors.lastname: Blagitko-Dorfs
authors.affiliation: Department of Internal Medicine I, University Freiburg-Medical Center, Faculty of Medicine, Freiburg, Germany.
authors.forename: Christine
authors.initials: C
authors.lastname: Greil
authors.affiliation: Department of Pathology and Tumor Biology, Graduate School of Medicine, Kyoto University, Kyoto, Japan.
authors.forename: Kenichi
authors.initials: K
authors.lastname: Yoshida
authors.affiliation: Department of Internal Medicine I, University Freiburg-Medical Center, Faculty of Medicine, Freiburg, Germany.
authors.forename: Dietmar
authors.initials: D
authors.lastname: Pfeifer
authors.affiliation: Department of Internal Medicine I, University Freiburg-Medical Center, Faculty of Medicine, Freiburg, Germany.
authors.forename: Marie
authors.initials: M
authors.lastname: Follo
authors.affiliation: Department of Internal Medicine I, University Freiburg-Medical Center, Faculty of Medicine, Freiburg, Germany.
authors.forename: Justus
authors.initials: J
authors.lastname: Duyster
authors.affiliation: Department of Internal Medicine I, University Freiburg-Medical Center, Faculty of Medicine, Freiburg, Germany.
authors.forename: Rainer
authors.initials: R
authors.lastname: Claus
authors.affiliation: Department of Pathology and Tumor Biology, Graduate School of Medicine, Kyoto University, Kyoto, Japan.
authors.forename: Seishi
authors.initials: S
authors.lastname: Ogawa
authors.affiliation: Department of Internal Medicine I, University Freiburg-Medical Center, Faculty of Medicine, Freiburg, Germany.
authors.forename: Michael
authors.initials: M
authors.lastname: Lübbert
authors.affiliation: Department of Internal Medicine I, University Freiburg-Medical Center, Faculty of Medicine, Freiburg, Germany. Electronic address: heiko.becker@uniklinik-freiburg.de.
authors.forename: Heiko
authors.initials: H
authors.lastname: Becker
mesh_terms: D049109:Cell Proliferation; D002999:Clone Cells / Q000473:pathology; D059472:Exome / Q000235:genetics*; D005838:Genotype; D020662:Guanine Nucleotide Exchange Factors / Q000235:genetics; D006410:Hematopoiesis; D015470:Leukemia, Myeloid, Acute / Q000209:etiology / Q000235:genetics* / Q000473:pathology; D009154:Mutation; D015514:Oncogene Proteins, Fusion; D054623:Receptor-Like Protein Tyrosine Phosphatases, Class 2 / Q000235:genetics; D059010:Single-Cell Analysis / Q000379:methods*; D014157:Transcription Factors / Q000235:genetics
chemical_list: C486317:CAND1 protein, human; C000607938:DOCK6 protein, human; D020662:Guanine Nucleotide Exchange Factors; D015514:Oncogene Proteins, Fusion; D014157:Transcription Factors; C471254:inv(16) fusion protein, human; C516865:PTPRT protein, human; D054623:Receptor-Like Protein Tyrosine Phosphatases, Class 2
keywords: AML; CBL; Clonal hematopoiesis; PTPRT; Single cell; inv(16)