Clinical validation of an ultra high-throughput spiral microfluidics for the detection and enrichment of viable circulating tumor cells.

<h4>Background</h4>Circulating tumor cells (CTCs) are cancer cells that can be isolated via liquid biopsy from blood and can be phenotypically and genetically characterized to provide critical information for guiding cancer treatment. Current analysis of CTCs is hindered by the throughpu...

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Autores principales: Bee Luan Khoo, Majid Ebrahimi Warkiani, Daniel Shao-Weng Tan, Ali Asgar S Bhagat, Darryl Irwin, Dawn Pingxi Lau, Alvin S T Lim, Kiat Hon Lim, Sai Sakktee Krisna, Wan-Teck Lim, Yoon Sim Yap, Soo Chin Lee, Ross A Soo, Jongyoon Han, Chwee Teck Lim
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Publicado: Public Library of Science (PLoS) 2014
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spelling oai:doaj.org-article:03951ce56787415fbd46f96bac1202e52021-11-25T06:09:33ZClinical validation of an ultra high-throughput spiral microfluidics for the detection and enrichment of viable circulating tumor cells.1932-620310.1371/journal.pone.0099409https://doaj.org/article/03951ce56787415fbd46f96bac1202e52014-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/24999991/?tool=EBIhttps://doaj.org/toc/1932-6203<h4>Background</h4>Circulating tumor cells (CTCs) are cancer cells that can be isolated via liquid biopsy from blood and can be phenotypically and genetically characterized to provide critical information for guiding cancer treatment. Current analysis of CTCs is hindered by the throughput, selectivity and specificity of devices or assays used in CTC detection and isolation.<h4>Methodology/principal findings</h4>Here, we enriched and characterized putative CTCs from blood samples of patients with both advanced stage metastatic breast and lung cancers using a novel multiplexed spiral microfluidic chip. This system detected putative CTCs under high sensitivity (100%, n = 56) (Breast cancer samples: 12-1275 CTCs/ml; Lung cancer samples: 10-1535 CTCs/ml) rapidly from clinically relevant blood volumes (7.5 ml under 5 min). Blood samples were completely separated into plasma, CTCs and PBMCs components and each fraction were characterized with immunophenotyping (Pan-cytokeratin/CD45, CD44/CD24, EpCAM), fluorescence in-situ hybridization (FISH) (EML4-ALK) or targeted somatic mutation analysis. We used an ultra-sensitive mass spectrometry based system to highlight the presence of an EGFR-activating mutation in both isolated CTCs and plasma cell-free DNA (cf-DNA), and demonstrate concordance with the original tumor-biopsy samples.<h4>Conclusions/significance</h4>We have clinically validated our multiplexed microfluidic chip for the ultra high-throughput, low-cost and label-free enrichment of CTCs. Retrieved cells were unlabeled and viable, enabling potential propagation and real-time downstream analysis using next generation sequencing (NGS) or proteomic analysis.Bee Luan KhooMajid Ebrahimi WarkianiDaniel Shao-Weng TanAli Asgar S BhagatDarryl IrwinDawn Pingxi LauAlvin S T LimKiat Hon LimSai Sakktee KrisnaWan-Teck LimYoon Sim YapSoo Chin LeeRoss A SooJongyoon HanChwee Teck LimPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 9, Iss 7, p e99409 (2014)
institution DOAJ
collection DOAJ
language EN
topic Medicine
R
Science
Q
spellingShingle Medicine
R
Science
Q
Bee Luan Khoo
Majid Ebrahimi Warkiani
Daniel Shao-Weng Tan
Ali Asgar S Bhagat
Darryl Irwin
Dawn Pingxi Lau
Alvin S T Lim
Kiat Hon Lim
Sai Sakktee Krisna
Wan-Teck Lim
Yoon Sim Yap
Soo Chin Lee
Ross A Soo
Jongyoon Han
Chwee Teck Lim
Clinical validation of an ultra high-throughput spiral microfluidics for the detection and enrichment of viable circulating tumor cells.
description <h4>Background</h4>Circulating tumor cells (CTCs) are cancer cells that can be isolated via liquid biopsy from blood and can be phenotypically and genetically characterized to provide critical information for guiding cancer treatment. Current analysis of CTCs is hindered by the throughput, selectivity and specificity of devices or assays used in CTC detection and isolation.<h4>Methodology/principal findings</h4>Here, we enriched and characterized putative CTCs from blood samples of patients with both advanced stage metastatic breast and lung cancers using a novel multiplexed spiral microfluidic chip. This system detected putative CTCs under high sensitivity (100%, n = 56) (Breast cancer samples: 12-1275 CTCs/ml; Lung cancer samples: 10-1535 CTCs/ml) rapidly from clinically relevant blood volumes (7.5 ml under 5 min). Blood samples were completely separated into plasma, CTCs and PBMCs components and each fraction were characterized with immunophenotyping (Pan-cytokeratin/CD45, CD44/CD24, EpCAM), fluorescence in-situ hybridization (FISH) (EML4-ALK) or targeted somatic mutation analysis. We used an ultra-sensitive mass spectrometry based system to highlight the presence of an EGFR-activating mutation in both isolated CTCs and plasma cell-free DNA (cf-DNA), and demonstrate concordance with the original tumor-biopsy samples.<h4>Conclusions/significance</h4>We have clinically validated our multiplexed microfluidic chip for the ultra high-throughput, low-cost and label-free enrichment of CTCs. Retrieved cells were unlabeled and viable, enabling potential propagation and real-time downstream analysis using next generation sequencing (NGS) or proteomic analysis.
format article
author Bee Luan Khoo
Majid Ebrahimi Warkiani
Daniel Shao-Weng Tan
Ali Asgar S Bhagat
Darryl Irwin
Dawn Pingxi Lau
Alvin S T Lim
Kiat Hon Lim
Sai Sakktee Krisna
Wan-Teck Lim
Yoon Sim Yap
Soo Chin Lee
Ross A Soo
Jongyoon Han
Chwee Teck Lim
author_facet Bee Luan Khoo
Majid Ebrahimi Warkiani
Daniel Shao-Weng Tan
Ali Asgar S Bhagat
Darryl Irwin
Dawn Pingxi Lau
Alvin S T Lim
Kiat Hon Lim
Sai Sakktee Krisna
Wan-Teck Lim
Yoon Sim Yap
Soo Chin Lee
Ross A Soo
Jongyoon Han
Chwee Teck Lim
author_sort Bee Luan Khoo
title Clinical validation of an ultra high-throughput spiral microfluidics for the detection and enrichment of viable circulating tumor cells.
title_short Clinical validation of an ultra high-throughput spiral microfluidics for the detection and enrichment of viable circulating tumor cells.
title_full Clinical validation of an ultra high-throughput spiral microfluidics for the detection and enrichment of viable circulating tumor cells.
title_fullStr Clinical validation of an ultra high-throughput spiral microfluidics for the detection and enrichment of viable circulating tumor cells.
title_full_unstemmed Clinical validation of an ultra high-throughput spiral microfluidics for the detection and enrichment of viable circulating tumor cells.
title_sort clinical validation of an ultra high-throughput spiral microfluidics for the detection and enrichment of viable circulating tumor cells.
publisher Public Library of Science (PLoS)
publishDate 2014
url https://doaj.org/article/03951ce56787415fbd46f96bac1202e5
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