title: Genome-wide quantification of rare somatic mutations in normal human tissues using massively parallel sequencing.
abstract: We present the bottleneck sequencing system (BotSeqS), a next-generation sequencing method that simultaneously quantifies rare somatic point mutations across the mitochondrial and nuclear genomes. BotSeqS combines molecular barcoding with a simple dilution step immediately before library amplification. We use BotSeqS to show age- and tissue-dependent accumulations of rare mutations and demonstrate that somatic mutational burden in normal human tissues can vary by several orders of magnitude, depending on biologic and environmental factors. We further show major differences between the mutational patterns of the mitochondrial and nuclear genomes in normal tissues. Lastly, the mutation spectra of normal tissues were different from each other, but similar to those of the cancers that arose in them. This technology can provide insights into the number and nature of genetic alterations in normal tissues and can be used to address a variety of fundamental questions about the genomes of diseased tissues.
authors.affiliation: Ludwig Center for Cancer Genetics and Therapeutics, Department of Oncology, Johns Hopkins Kimmel Cancer Center, Baltimore, MD 21287; The Howard Hughes Medical Institute, Johns Hopkins University, Baltimore, MD 21231;
authors.forename: Margaret L
authors.initials: ML
authors.lastname: Hoang
authors.affiliation: Ludwig Center for Cancer Genetics and Therapeutics, Department of Oncology, Johns Hopkins Kimmel Cancer Center, Baltimore, MD 21287; The Howard Hughes Medical Institute, Johns Hopkins University, Baltimore, MD 21231;
authors.forename: Isaac
authors.initials: I
authors.lastname: Kinde
authors.affiliation: Division of Biostatistics and Bioinformatics, Department of Oncology, Johns Hopkins Kimmel Cancer Center, Baltimore, MD 21205; Department of Biostatistics, Johns Hopkins Bloomberg School of Public Health, Baltimore, MD 21205;
authors.forename: Cristian
authors.initials: C
authors.lastname: Tomasetti
authors.affiliation: Ludwig Center for Cancer Genetics and Therapeutics, Department of Oncology, Johns Hopkins Kimmel Cancer Center, Baltimore, MD 21287; The Howard Hughes Medical Institute, Johns Hopkins University, Baltimore, MD 21231;
authors.forename: K Wyatt
authors.initials: KW
authors.lastname: McMahon
authors.affiliation: Department of Pharmacological Sciences, Stony Brook University, Stony Brook, NY 11794;
authors.forename: Thomas A
authors.initials: TA
authors.lastname: Rosenquist
authors.affiliation: Department of Pharmacological Sciences, Stony Brook University, Stony Brook, NY 11794; Department of Medicine, Stony Brook University, Stony Brook, NY 11794;
authors.forename: Arthur P
authors.initials: AP
authors.lastname: Grollman
authors.affiliation: Ludwig Center for Cancer Genetics and Therapeutics, Department of Oncology, Johns Hopkins Kimmel Cancer Center, Baltimore, MD 21287; bertvog@gmail.com kinzlke@jhmi.edu.
authors.forename: Kenneth W
authors.initials: KW
authors.lastname: Kinzler
authors.affiliation: Ludwig Center for Cancer Genetics and Therapeutics, Department of Oncology, Johns Hopkins Kimmel Cancer Center, Baltimore, MD 21287; The Howard Hughes Medical Institute, Johns Hopkins University, Baltimore, MD 21231; Department of Pathology, Johns Hopkins School of Medicine, Baltimore, MD 21231 bertvog@gmail.com kinzlke@jhmi.edu.
authors.forename: Bert
authors.initials: B
authors.lastname: Vogelstein
authors.affiliation: Ludwig Center for Cancer Genetics and Therapeutics, Department of Oncology, Johns Hopkins Kimmel Cancer Center, Baltimore, MD 21287; Department of Pathology, Johns Hopkins School of Medicine, Baltimore, MD 21231.
authors.forename: Nickolas
authors.initials: N
authors.lastname: Papadopoulos
mesh_terms: D000293:Adolescent; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D002467:Cell Nucleus / Q000235:genetics; D002648:Child; D002675:Child, Preschool; D004272:DNA, Mitochondrial / Q000737:chemistry / Q000235:genetics; D005260:Female; D015894:Genome, Human / Q000235:genetics*; D023281:Genomics / Q000379:methods*; D059014:High-Throughput Nucleotide Sequencing / Q000379:methods*; D006801:Humans; D008297:Male; D008875:Middle Aged; D009154:Mutation; D055815:Young Adult
chemical_list: D004272:DNA, Mitochondrial
keywords: aging; genomics; next-generation sequencing; somatic mutation