Single-cell DNA and RNA sequencing of circulating tumor cells

Abstract Single-cell sequencing of circulating tumor cells can precisely represent tumor heterogeneity and provide useful information for cancer treatment and research. After spiking TGW neuroblastoma cells into blood derived from healthy volunteer, the cells were isolated by fluorescence-activated...

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Autores principales:	Masato Kojima, Takanori Harada, Takahiro Fukazawa, Sho Kurihara, Isamu Saeki, Shinya Takahashi, Eiso Hiyama
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/8ca0c1b239af4bb3ae012c967c0c0408
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spelling	oai:doaj.org-article:8ca0c1b239af4bb3ae012c967c0c04082021-11-28T12:17:58ZSingle-cell DNA and RNA sequencing of circulating tumor cells10.1038/s41598-021-02165-72045-2322https://doaj.org/article/8ca0c1b239af4bb3ae012c967c0c04082021-11-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-02165-7https://doaj.org/toc/2045-2322Abstract Single-cell sequencing of circulating tumor cells can precisely represent tumor heterogeneity and provide useful information for cancer treatment and research. After spiking TGW neuroblastoma cells into blood derived from healthy volunteer, the cells were isolated by fluorescence-activated cell sorting. DNA and mRNA were amplified by four different whole-genome amplifications (WGA) and three whole-transcriptome amplifications (WTA) methods, followed by single-cell DNA and RNA sequencing. Multiple displacement amplification (MDA)-based WGA methods showed higher amplification efficiency than other methods with a comparable depth of coverage as the bulk sample. The uniformity of coverage greatly differed among samples (12.5–89.2%), with some samples evaluated by the MDA-based WGA method using phi29 DNA polymerase and random primers showing a high (> 80%) uniformity of coverage. The MDA-based WTA method less effectively amplified mRNA and showed non-specific gene expression patterns. The PCR-based WTA using template switching with locked nucleic acid technology accurately amplified mRNA from a single cell. Taken together, our results present a more reliable and adaptable approach for CTC profiling at the single-cell level. Such molecular information on CTCs derived from clinical patients will promote cancer treatment and research.Masato KojimaTakanori HaradaTakahiro FukazawaSho KuriharaIsamu SaekiShinya TakahashiEiso HiyamaNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-8 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Masato Kojima Takanori Harada Takahiro Fukazawa Sho Kurihara Isamu Saeki Shinya Takahashi Eiso Hiyama Single-cell DNA and RNA sequencing of circulating tumor cells
description	Abstract Single-cell sequencing of circulating tumor cells can precisely represent tumor heterogeneity and provide useful information for cancer treatment and research. After spiking TGW neuroblastoma cells into blood derived from healthy volunteer, the cells were isolated by fluorescence-activated cell sorting. DNA and mRNA were amplified by four different whole-genome amplifications (WGA) and three whole-transcriptome amplifications (WTA) methods, followed by single-cell DNA and RNA sequencing. Multiple displacement amplification (MDA)-based WGA methods showed higher amplification efficiency than other methods with a comparable depth of coverage as the bulk sample. The uniformity of coverage greatly differed among samples (12.5–89.2%), with some samples evaluated by the MDA-based WGA method using phi29 DNA polymerase and random primers showing a high (> 80%) uniformity of coverage. The MDA-based WTA method less effectively amplified mRNA and showed non-specific gene expression patterns. The PCR-based WTA using template switching with locked nucleic acid technology accurately amplified mRNA from a single cell. Taken together, our results present a more reliable and adaptable approach for CTC profiling at the single-cell level. Such molecular information on CTCs derived from clinical patients will promote cancer treatment and research.
format	article
author	Masato Kojima Takanori Harada Takahiro Fukazawa Sho Kurihara Isamu Saeki Shinya Takahashi Eiso Hiyama
author_facet	Masato Kojima Takanori Harada Takahiro Fukazawa Sho Kurihara Isamu Saeki Shinya Takahashi Eiso Hiyama
author_sort	Masato Kojima
title	Single-cell DNA and RNA sequencing of circulating tumor cells
title_short	Single-cell DNA and RNA sequencing of circulating tumor cells
title_full	Single-cell DNA and RNA sequencing of circulating tumor cells
title_fullStr	Single-cell DNA and RNA sequencing of circulating tumor cells
title_full_unstemmed	Single-cell DNA and RNA sequencing of circulating tumor cells
title_sort	single-cell dna and rna sequencing of circulating tumor cells
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/8ca0c1b239af4bb3ae012c967c0c0408
work_keys_str_mv	AT masatokojima singlecelldnaandrnasequencingofcirculatingtumorcells AT takanoriharada singlecelldnaandrnasequencingofcirculatingtumorcells AT takahirofukazawa singlecelldnaandrnasequencingofcirculatingtumorcells AT shokurihara singlecelldnaandrnasequencingofcirculatingtumorcells AT isamusaeki singlecelldnaandrnasequencingofcirculatingtumorcells AT shinyatakahashi singlecelldnaandrnasequencingofcirculatingtumorcells AT eisohiyama singlecelldnaandrnasequencingofcirculatingtumorcells
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Single-cell DNA and RNA sequencing of circulating tumor cells

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