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Antigen-agnostic microfluidics-based circulating tumor cell enrichment and downstream molecular characterization

Circulating tumor cells (CTC) isolated from the peripheral blood of cancer patients by a minimally invasive procedure provide surrogate markers of the tumor that can be repeatedly sampled. However, the selection and enumeration of CTCs by traditional methods based on surface proteins like EPCAM may...

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Autores principales: Cohen, Evan N., Jayachandran, Gitanjali, Hardy, Max R., Venkata Subramanian, Ananya M., Meng, Xiangtian, Reuben, James M.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7584183/
https://www.ncbi.nlm.nih.gov/pubmed/33095819
http://dx.doi.org/10.1371/journal.pone.0241123
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author Cohen, Evan N.
Jayachandran, Gitanjali
Hardy, Max R.
Venkata Subramanian, Ananya M.
Meng, Xiangtian
Reuben, James M.
author_facet Cohen, Evan N.
Jayachandran, Gitanjali
Hardy, Max R.
Venkata Subramanian, Ananya M.
Meng, Xiangtian
Reuben, James M.
author_sort Cohen, Evan N.
collection PubMed
description Circulating tumor cells (CTC) isolated from the peripheral blood of cancer patients by a minimally invasive procedure provide surrogate markers of the tumor that can be repeatedly sampled. However, the selection and enumeration of CTCs by traditional methods based on surface proteins like EPCAM may not detect CTCs with a mesenchymal phenotype. Here, we employed an antibody-agnostic platform, the Parsortix(®) PR1 system, which enriches CTCs based on cell size and membrane deformability. We evaluated the linearity, sensitivity, and specificity of the Parsortix PR1 system in tandem with 3 downstream molecular characterization techniques using healthy donor blood spiked with cultured cell lines. Signal amplification of mRNA using a QuantiGene 25-gene assay was able to quantitate multiple epithelial genes, including CDH1, EGFR, ERBB2, KRT18, and MUC1, from high numbers of spiked cells and was able to detect KRT18 when only 50 MCF-7 or SUM190 cells were spiked into healthy donor blood. However, target amplification of mRNA by quantitative polymerase chain reaction (qPCR) showed better sensitivity; qPCR without pre-amplification was able to detect CTC-related genes in Parsortix PR1-enriched cells when as few as 5 SKBR3 cells were spiked into blood. Finally, the HTG EdgeSeq nuclease protection assay was able to profile mRNA expression of over 2,560 cancer-related genes from Parsortix PR1 enriched cells, showing enrichment in cancer signaling pathways and ERBB2, KRT19, and KRT7. Overall, the Parsortix PR1 platform may be amenable to transition into routine clinical workflows.
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spelling pubmed-75841832020-10-27 Antigen-agnostic microfluidics-based circulating tumor cell enrichment and downstream molecular characterization Cohen, Evan N. Jayachandran, Gitanjali Hardy, Max R. Venkata Subramanian, Ananya M. Meng, Xiangtian Reuben, James M. PLoS One Research Article Circulating tumor cells (CTC) isolated from the peripheral blood of cancer patients by a minimally invasive procedure provide surrogate markers of the tumor that can be repeatedly sampled. However, the selection and enumeration of CTCs by traditional methods based on surface proteins like EPCAM may not detect CTCs with a mesenchymal phenotype. Here, we employed an antibody-agnostic platform, the Parsortix(®) PR1 system, which enriches CTCs based on cell size and membrane deformability. We evaluated the linearity, sensitivity, and specificity of the Parsortix PR1 system in tandem with 3 downstream molecular characterization techniques using healthy donor blood spiked with cultured cell lines. Signal amplification of mRNA using a QuantiGene 25-gene assay was able to quantitate multiple epithelial genes, including CDH1, EGFR, ERBB2, KRT18, and MUC1, from high numbers of spiked cells and was able to detect KRT18 when only 50 MCF-7 or SUM190 cells were spiked into healthy donor blood. However, target amplification of mRNA by quantitative polymerase chain reaction (qPCR) showed better sensitivity; qPCR without pre-amplification was able to detect CTC-related genes in Parsortix PR1-enriched cells when as few as 5 SKBR3 cells were spiked into blood. Finally, the HTG EdgeSeq nuclease protection assay was able to profile mRNA expression of over 2,560 cancer-related genes from Parsortix PR1 enriched cells, showing enrichment in cancer signaling pathways and ERBB2, KRT19, and KRT7. Overall, the Parsortix PR1 platform may be amenable to transition into routine clinical workflows. Public Library of Science 2020-10-23 /pmc/articles/PMC7584183/ /pubmed/33095819 http://dx.doi.org/10.1371/journal.pone.0241123 Text en © 2020 Cohen et al http://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Article
Cohen, Evan N.
Jayachandran, Gitanjali
Hardy, Max R.
Venkata Subramanian, Ananya M.
Meng, Xiangtian
Reuben, James M.
Antigen-agnostic microfluidics-based circulating tumor cell enrichment and downstream molecular characterization
title Antigen-agnostic microfluidics-based circulating tumor cell enrichment and downstream molecular characterization
title_full Antigen-agnostic microfluidics-based circulating tumor cell enrichment and downstream molecular characterization
title_fullStr Antigen-agnostic microfluidics-based circulating tumor cell enrichment and downstream molecular characterization
title_full_unstemmed Antigen-agnostic microfluidics-based circulating tumor cell enrichment and downstream molecular characterization
title_short Antigen-agnostic microfluidics-based circulating tumor cell enrichment and downstream molecular characterization
title_sort antigen-agnostic microfluidics-based circulating tumor cell enrichment and downstream molecular characterization
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7584183/
https://www.ncbi.nlm.nih.gov/pubmed/33095819
http://dx.doi.org/10.1371/journal.pone.0241123
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