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mRNA-Seq of single prostate cancer circulating tumor cells reveals recapitulation of gene expression and pathways found in prostate cancer
Circulating tumor cells (CTC) mediate metastatic spread of many solid tumors and enumeration of CTCs is currently used as a prognostic indicator of survival in metastatic prostate cancer patients. Some evidence suggests that it is possible to derive additional information about tumors from expressio...
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Published in: | PloS one 2012-11, Vol.7 (11), p.e49144-e49144 |
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creator | Cann, Gordon M Gulzar, Zulfiqar G Cooper, Samantha Li, Robin Luo, Shujun Tat, Mai Stuart, Sarah Schroth, Gary Srinivas, Sandhya Ronaghi, Mostafa Brooks, James D Talasaz, Amirali H |
description | Circulating tumor cells (CTC) mediate metastatic spread of many solid tumors and enumeration of CTCs is currently used as a prognostic indicator of survival in metastatic prostate cancer patients. Some evidence suggests that it is possible to derive additional information about tumors from expression analysis of CTCs, but the technical difficulty of isolating and analyzing individual CTCs has limited progress in this area. To assess the ability of a new generation of MagSweeper to isolate intact CTCs for downstream analysis, we performed mRNA-Seq on single CTCs isolated from the blood of patients with metastatic prostate cancer and on single prostate cancer cell line LNCaP cells spiked into the blood of healthy donors. We found that the MagSweeper effectively isolated CTCs with a capture efficiency that matched the CellSearch platform. However, unlike CellSearch, the MagSweeper facilitates isolation of individual live CTCs without contaminating leukocytes. Importantly, mRNA-Seq analysis showed that the MagSweeper isolation process did not have a discernible impact on the transcriptional profile of single LNCaPs isolated from spiked human blood, suggesting that any perturbations caused by the MagSweeper process on the transcriptional signature of isolated cells are modest. Although the RNA from patient CTCs showed signs of significant degradation, consistent with reports of short half-lives and apoptosis amongst CTCs, transcriptional signatures of prostate tissue and of cancer were readily detectable with single CTC mRNA-Seq. These results demonstrate that the MagSweeper provides access to intact CTCs and that these CTCs can potentially supply clinically relevant information. |
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Some evidence suggests that it is possible to derive additional information about tumors from expression analysis of CTCs, but the technical difficulty of isolating and analyzing individual CTCs has limited progress in this area. To assess the ability of a new generation of MagSweeper to isolate intact CTCs for downstream analysis, we performed mRNA-Seq on single CTCs isolated from the blood of patients with metastatic prostate cancer and on single prostate cancer cell line LNCaP cells spiked into the blood of healthy donors. We found that the MagSweeper effectively isolated CTCs with a capture efficiency that matched the CellSearch platform. However, unlike CellSearch, the MagSweeper facilitates isolation of individual live CTCs without contaminating leukocytes. Importantly, mRNA-Seq analysis showed that the MagSweeper isolation process did not have a discernible impact on the transcriptional profile of single LNCaPs isolated from spiked human blood, suggesting that any perturbations caused by the MagSweeper process on the transcriptional signature of isolated cells are modest. Although the RNA from patient CTCs showed signs of significant degradation, consistent with reports of short half-lives and apoptosis amongst CTCs, transcriptional signatures of prostate tissue and of cancer were readily detectable with single CTC mRNA-Seq. These results demonstrate that the MagSweeper provides access to intact CTCs and that these CTCs can potentially supply clinically relevant information.</description><identifier>ISSN: 1932-6203</identifier><identifier>EISSN: 1932-6203</identifier><identifier>DOI: 10.1371/journal.pone.0049144</identifier><identifier>PMID: 23145101</identifier><language>eng</language><publisher>United States: Public Library of Science</publisher><subject>Analysis ; Apoptosis ; Biology ; Biomarkers, Tumor - blood ; Blood ; Cancer ; Cancer genetics ; Cancer metastasis ; Cell Line, Tumor ; Engineering ; Enumeration ; Gene expression ; Gene Expression Regulation, Neoplastic ; Human subjects ; Humans ; Kinases ; Leukocytes ; Male ; Medicine ; Messenger RNA ; Metabolic Networks and Pathways ; Metastases ; Metastasis ; Neoplasm Metastasis - genetics ; Neoplasm Metastasis - pathology ; Neoplastic Cells, Circulating - metabolism ; Neoplastic Cells, Circulating - pathology ; Patients ; Prognosis ; Prostate cancer ; Prostatic Neoplasms - blood ; Prostatic Neoplasms - genetics ; Prostatic Neoplasms - pathology ; Ribonucleic acid ; RNA ; RNA, Messenger - blood ; RNA, Messenger - genetics ; Sequence Alignment ; Sequence Analysis, RNA ; Solid tumors ; Transcription ; Transcription (Genetics) ; Tumor cells ; Tumors ; Urology</subject><ispartof>PloS one, 2012-11, Vol.7 (11), p.e49144-e49144</ispartof><rights>COPYRIGHT 2012 Public Library of Science</rights><rights>2012 Cann et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License: https://creativecommons.org/licenses/by/4.0/ (the “License”), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. 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These results demonstrate that the MagSweeper provides access to intact CTCs and that these CTCs can potentially supply clinically relevant information.</description><subject>Analysis</subject><subject>Apoptosis</subject><subject>Biology</subject><subject>Biomarkers, Tumor - blood</subject><subject>Blood</subject><subject>Cancer</subject><subject>Cancer genetics</subject><subject>Cancer metastasis</subject><subject>Cell Line, Tumor</subject><subject>Engineering</subject><subject>Enumeration</subject><subject>Gene expression</subject><subject>Gene Expression Regulation, Neoplastic</subject><subject>Human subjects</subject><subject>Humans</subject><subject>Kinases</subject><subject>Leukocytes</subject><subject>Male</subject><subject>Medicine</subject><subject>Messenger RNA</subject><subject>Metabolic Networks and Pathways</subject><subject>Metastases</subject><subject>Metastasis</subject><subject>Neoplasm Metastasis - genetics</subject><subject>Neoplasm Metastasis - pathology</subject><subject>Neoplastic Cells, Circulating - 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Performed the experiments: GMC ZGG RL SL MT SS. Analyzed the data: SC AT GMC ZGG MR JDB. Contributed reagents/materials/analysis tools: GS SS JDB AT. Wrote the paper: GMC AT ZGG JDB.</notes><notes>Competing Interests: The authors have read the journal's policy and have the following conflicts: GMC, SC, RL, SL, MT, SS, GS, MR, and AT are currently employed by Illumina. This does not alter the authors' adherence to all the PLOS ONE policies on sharing data and materials.</notes><abstract>Circulating tumor cells (CTC) mediate metastatic spread of many solid tumors and enumeration of CTCs is currently used as a prognostic indicator of survival in metastatic prostate cancer patients. Some evidence suggests that it is possible to derive additional information about tumors from expression analysis of CTCs, but the technical difficulty of isolating and analyzing individual CTCs has limited progress in this area. To assess the ability of a new generation of MagSweeper to isolate intact CTCs for downstream analysis, we performed mRNA-Seq on single CTCs isolated from the blood of patients with metastatic prostate cancer and on single prostate cancer cell line LNCaP cells spiked into the blood of healthy donors. We found that the MagSweeper effectively isolated CTCs with a capture efficiency that matched the CellSearch platform. However, unlike CellSearch, the MagSweeper facilitates isolation of individual live CTCs without contaminating leukocytes. Importantly, mRNA-Seq analysis showed that the MagSweeper isolation process did not have a discernible impact on the transcriptional profile of single LNCaPs isolated from spiked human blood, suggesting that any perturbations caused by the MagSweeper process on the transcriptional signature of isolated cells are modest. Although the RNA from patient CTCs showed signs of significant degradation, consistent with reports of short half-lives and apoptosis amongst CTCs, transcriptional signatures of prostate tissue and of cancer were readily detectable with single CTC mRNA-Seq. These results demonstrate that the MagSweeper provides access to intact CTCs and that these CTCs can potentially supply clinically relevant information.</abstract><cop>United States</cop><pub>Public Library of Science</pub><pmid>23145101</pmid><doi>10.1371/journal.pone.0049144</doi><tpages>e49144</tpages><oa>free_for_read</oa></addata></record> |
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recordid | cdi_plos_journals_1326733351 |
source | PMC (PubMed Central); Publicly Available Content (ProQuest) |
subjects | Analysis Apoptosis Biology Biomarkers, Tumor - blood Blood Cancer Cancer genetics Cancer metastasis Cell Line, Tumor Engineering Enumeration Gene expression Gene Expression Regulation, Neoplastic Human subjects Humans Kinases Leukocytes Male Medicine Messenger RNA Metabolic Networks and Pathways Metastases Metastasis Neoplasm Metastasis - genetics Neoplasm Metastasis - pathology Neoplastic Cells, Circulating - metabolism Neoplastic Cells, Circulating - pathology Patients Prognosis Prostate cancer Prostatic Neoplasms - blood Prostatic Neoplasms - genetics Prostatic Neoplasms - pathology Ribonucleic acid RNA RNA, Messenger - blood RNA, Messenger - genetics Sequence Alignment Sequence Analysis, RNA Solid tumors Transcription Transcription (Genetics) Tumor cells Tumors Urology |
title | mRNA-Seq of single prostate cancer circulating tumor cells reveals recapitulation of gene expression and pathways found in prostate cancer |
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