Show simple item record

dc.contributor.authorFerrarini, Aen_US
dc.contributor.authorForcato, Cen_US
dc.contributor.authorBuson, Gen_US
dc.contributor.authorTononi, Pen_US
dc.contributor.authorDel Monaco, Ven_US
dc.contributor.authorTerracciano, Men_US
dc.contributor.authorBolognesi, Cen_US
dc.contributor.authorFontana, Fen_US
dc.contributor.authorMedoro, Gen_US
dc.contributor.authorNeves, Ren_US
dc.contributor.authorMöhlendick, Ben_US
dc.contributor.authorRihawi, Ken_US
dc.contributor.authorArdizzoni, Aen_US
dc.contributor.authorSumanasuriya, Sen_US
dc.contributor.authorFlohr, Pen_US
dc.contributor.authorLambros, Men_US
dc.contributor.authorde Bono, Jen_US
dc.contributor.authorStoecklein, NHen_US
dc.contributor.authorManaresi, Nen_US
dc.coverage.spatialUnited Statesen_US
dc.identifier.citationPLoS One, 2018, 13 (3), pp. e0193689 - ?en_US
dc.description.abstractChromosomal instability and associated chromosomal aberrations are hallmarks of cancer and play a critical role in disease progression and development of resistance to drugs. Single-cell genome analysis has gained interest in latest years as a source of biomarkers for targeted-therapy selection and drug resistance, and several methods have been developed to amplify the genomic DNA and to produce libraries suitable for Whole Genome Sequencing (WGS). However, most protocols require several enzymatic and cleanup steps, thus increasing the complexity and length of protocols, while robustness and speed are key factors for clinical applications. To tackle this issue, we developed a single-tube, single-step, streamlined protocol, exploiting ligation mediated PCR (LM-PCR) Whole Genome Amplification (WGA) method, for low-pass genome sequencing with the Ion Torrent™ platform and copy number alterations (CNAs) calling from single cells. The method was evaluated on single cells isolated from 6 aberrant cell lines of the NCI-H series. In addition, to demonstrate the feasibility of the workflow on clinical samples, we analyzed single circulating tumor cells (CTCs) and white blood cells (WBCs) isolated from the blood of patients affected by prostate cancer or lung adenocarcinoma. The results obtained show that the developed workflow generates data accurately representing whole genome absolute copy number profiles of single cell and allows alterations calling at resolutions down to 100 Kbp with as few as 200,000 reads. The presented data demonstrate the feasibility of the Ampli1™ WGA-based low-pass workflow for detection of CNAs in single tumor cells which would be of particular interest for genome-driven targeted therapy selection and for monitoring of disease progression.en_US
dc.format.extente0193689 - ?en_US
dc.subjectAdenocarcinoma of Lungen_US
dc.subjectCell Line, Tumoren_US
dc.subjectDNA Copy Number Variationsen_US
dc.subjectHigh-Throughput Nucleotide Sequencingen_US
dc.subjectLung Neoplasmsen_US
dc.subjectNeoplastic Cells, Circulatingen_US
dc.subjectPolymerase Chain Reactionen_US
dc.subjectProstatic Neoplasmsen_US
dc.subjectSingle-Cell Analysisen_US
dc.subjectWhole Genome Sequencingen_US
dc.titleA streamlined workflow for single-cells genome-wide copy-number profiling by low-pass sequencing of LM-PCR whole-genome amplification products.en_US
dc.typeJournal Article
rioxxterms.typeJournal Article/Reviewen_US
dc.relation.isPartOfPLoS Oneen_US
pubs.notesNot knownen_US
pubs.organisational-group/ICR/Primary Group
pubs.organisational-group/ICR/Primary Group/ICR Divisions
pubs.organisational-group/ICR/Primary Group/ICR Divisions/Clinical Studies
pubs.organisational-group/ICR/Primary Group/ICR Divisions/Clinical Studies/Prostate Cancer Targeted Therapy Group
pubs.publication-statusPublished onlineen_US
pubs.embargo.termsNot knownen_US
icr.researchteamProstate Cancer Targeted Therapy Groupen_US
dc.contributor.icrauthorDe Bono, Johannen_US

Files in this item


This item appears in the following Collection(s)

Show simple item record
Except where otherwise noted, this item's license is described as