Clinical Validation of an Ultra High-Throughput Spiral Microfluidics for the Detection and Enrichment of Viable Circulating Tumor Cells
Background: 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...
| Main Authors: | , , , , , , , , , , , , , , |
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| Other Authors: | |
| Format: | Article |
| Language: | en_US |
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Public Library of Science
2014
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| Online Access: | http://hdl.handle.net/1721.1/89447 https://orcid.org/0000-0001-7215-1439 |
| _version_ | 1811088069830377472 |
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| author | Khoo, Bee Luan Warkiani, Majid Ebrahimi Tan, Daniel Shao-Weng Bhagat, Ali Asgar S. Irwin, Darryl Lau, Dawn Pingxi Lim, Alvin S. T. Lim, Kiat Hon Krisna, Sai Sakktee Lim, Wan-Teck Yap, Yoon Sim Lee, Soo Chin Soo, Ross A. Han, Jongyoon Lim, Chwee Teck |
| author2 | Massachusetts Institute of Technology. Department of Electrical Engineering and Computer Science |
| author_facet | Massachusetts Institute of Technology. Department of Electrical Engineering and Computer Science Khoo, Bee Luan Warkiani, Majid Ebrahimi Tan, Daniel Shao-Weng Bhagat, Ali Asgar S. Irwin, Darryl Lau, Dawn Pingxi Lim, Alvin S. T. Lim, Kiat Hon Krisna, Sai Sakktee Lim, Wan-Teck Yap, Yoon Sim Lee, Soo Chin Soo, Ross A. Han, Jongyoon Lim, Chwee Teck |
| author_sort | Khoo, Bee Luan |
| collection | MIT |
| description | Background:
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.
Methodology/Principal Findings:
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.
Conclusions/Significance:
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. |
| first_indexed | 2024-09-23T13:55:28Z |
| format | Article |
| id | mit-1721.1/89447 |
| institution | Massachusetts Institute of Technology |
| language | en_US |
| last_indexed | 2024-09-23T13:55:28Z |
| publishDate | 2014 |
| publisher | Public Library of Science |
| record_format | dspace |
| spelling | mit-1721.1/894472022-10-01T18:01:29Z Clinical Validation of an Ultra High-Throughput Spiral Microfluidics for the Detection and Enrichment of Viable Circulating Tumor Cells Khoo, Bee Luan Warkiani, Majid Ebrahimi Tan, Daniel Shao-Weng Bhagat, Ali Asgar S. Irwin, Darryl Lau, Dawn Pingxi Lim, Alvin S. T. Lim, Kiat Hon Krisna, Sai Sakktee Lim, Wan-Teck Yap, Yoon Sim Lee, Soo Chin Soo, Ross A. Han, Jongyoon Lim, Chwee Teck Massachusetts Institute of Technology. Department of Electrical Engineering and Computer Science Han, Jongyoon Background: 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. Methodology/Principal Findings: 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. Conclusions/Significance: 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. Singapore-MIT Alliance for Research and Technology 2014-09-11T18:20:53Z 2014-09-11T18:20:53Z 2014-07 2013-12 Article http://purl.org/eprint/type/JournalArticle 1932-6203 http://hdl.handle.net/1721.1/89447 Khoo, Bee Luan, Majid Ebrahimi Warkiani, Daniel Shao-Weng Tan, Ali Asgar S. Bhagat, Darryl Irwin, Dawn Pingxi Lau, Alvin S. T. Lim, et al. “Clinical Validation of an Ultra High-Throughput Spiral Microfluidics for the Detection and Enrichment of Viable Circulating Tumor Cells.” Edited by Hong Wanjin. PLoS ONE 9, no. 7 (July 7, 2014): e99409. https://orcid.org/0000-0001-7215-1439 en_US http://dx.doi.org/10.1371/journal.pone.0099409 PLoS ONE Creative Commons Attribution http://creativecommons.org/licenses/by/4.0/ application/pdf Public Library of Science Public Library of Science |
| spellingShingle | Khoo, Bee Luan Warkiani, Majid Ebrahimi Tan, Daniel Shao-Weng Bhagat, Ali Asgar S. Irwin, Darryl Lau, Dawn Pingxi Lim, Alvin S. T. Lim, Kiat Hon Krisna, Sai Sakktee Lim, Wan-Teck Yap, Yoon Sim Lee, Soo Chin Soo, Ross A. Han, Jongyoon Lim, Chwee Teck Clinical Validation of an Ultra High-Throughput Spiral Microfluidics for the Detection and Enrichment of Viable Circulating Tumor Cells |
| title | 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_short | 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 |
| url | http://hdl.handle.net/1721.1/89447 https://orcid.org/0000-0001-7215-1439 |
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