RT Journal
A1 Potter, Nicola E.
A1 Ermini, Luca
A1 Papaemmanuil, Elli
A1 Cazzaniga, Giovanni
A1 Vijayaraghavan, Gowri
A1 Titley, Ian
A1 Ford, Anthony
A1 Campbell, Peter
A1 Kearney, Lyndal
A1 Greaves, Mel
T1 Single-cell mutational profiling and clonal phylogeny in cancer
JF Genome Research 
JO Genome Research 
YR 2013 
FD December 01 
VO 23 
IS 12 
SP 2115 
OP 2125 
DO 10.1101/gr.159913.113 
UL http://genome.cshlp.org/content/23/12/2115.abstract 
AB The development of cancer is a dynamic evolutionary process in which intraclonal, genetic diversity provides a substrate for clonal selection and a source of therapeutic escape. The complexity and topography of intraclonal genetic architectures have major implications for biopsy-based prognosis and for targeted therapy. High-depth, next-generation sequencing (NGS) efficiently captures the mutational load of individual tumors or biopsies. But, being a snapshot portrait of total DNA, it disguises the fundamental features of subclonal variegation of genetic lesions and of clonal phylogeny. Single-cell genetic profiling provides a potential resolution to this problem, but methods developed to date all have limitations. We present a novel solution to this challenge using leukemic cells with known mutational spectra as a tractable model. DNA from flow-sorted single cells is screened using multiplex targeted Q-PCR within a microfluidic platform allowing unbiased single-cell selection, high-throughput, and comprehensive analysis for all main varieties of genetic abnormalities: chimeric gene fusions, copy number alterations, and single-nucleotide variants. We show, in this proof-of-principle study, that the method has a low error rate and can provide detailed subclonal genetic architectures and phylogenies.