Next-generation Sequencing (NGS) Analysis on Single Circulating Tumor Cells (CTCs) with No Need of Whole-genome Amplification (WGA)

Palmirotta, Raffaele; Lovero, Domenica; Silvestris, Erica; Felici, Claudia; Quaresmini, Davide; Cafforio, Paola; Silvestris, Franco

doi:10.21873/cgp.20029

Cited by 33 publications

(47 citation statements)

References 32 publications

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“…DEPArray™ is an advanced and highly automated instrument, which complements the capabilities of blood enrichment technologies to deliver pure individual cells suitable for an accurate molecular characterization. The versatility of DEPArray™ has been largely demonstrated by different research groups .…”

Section: Discussionmentioning

confidence: 99%

DEPArray™ system: An automatic image‐based sorter for isolation of pure circulating tumor cells

2018

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Circulating tumor cells (CTCs) are rare cells shed into the bloodstream by invasive tumors and their analysis offers a promising noninvasive tool to predict and monitor therapeutic responses. CTCs can be isolated from patient blood and their characterization at single‐cell level can inform on the genomic landscape of a tumor. All CTC enrichment methods bear a burden of contaminating normal cells, which mandate a further step of purification to enable reliable downstream genetic analysis. Here, we describe the DEPArray™ technology, a microchip‐based digital sorter, which combines precise microfluidic and microelectronic enabling precise, image‐based isolation of single CTCs, which can then be analyzed by Next Generation Sequencing (NGS) methods. © 2018 The Authors. Cytometry Part A published by Wiley Periodicals, Inc. on behalf of International Society for Advancement of Cytometry.

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Section: Discussionmentioning

confidence: 99%

DEPArray™ system: An automatic image‐based sorter for isolation of pure circulating tumor cells

2018

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“…Though, it has been optimized to work with highly fragmented and degraded FFPE DNA, recent studies have shown that it can be used to analyze mutations in cfDNA (https://www.illumina.com/). The Ion Ampliseq Colon and Lung cancer panel on the Ion Torrent Personal Genome Machine selectively amplifies 90 amplicons that encompass 1825 mutational hotspots of 22 genes related to colon and lung cancer [86][87][88]. It has been clinically validated in a retrospective study of 39 NSCLC samples and 51 colorectal cancer samples [84].…”

Section: Methods Targeting Druggable Mutation and Other Aberrations Imentioning

confidence: 99%

Liquid biopsy - emergence of a new era in personalized cancer care

2018

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The most successful treatment for cancer involves identifying druggable, biological markers for targeted therapy. In the clinical setting, surgical removal of tumors is the only procedure for identifying such targetable molecules. Shed from tumor cells, these markers are also present in circulating blood, albeit in very negligible amounts. Liquid biopsy is a procedure performed on a blood sample to look for such circulating cancer markers cells or pieces of nucleic acid from the tumor. The procedure shows promise in revolutionizing personalized cancer treatments. Here we briefly review the technique, characterization, and its utilization in clinics.

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“…By contrast, MDA methods have more advantages for SNVs for their high coverage breadth. More efforts should be devoted into developing bias‐free WGA methods or getting single‐genome sequencing without amplification . Zahn et al proposed a direct single‐cell whole‐genome library preparation method with high‐fidelity .…”

Section: Single Ctc Analysismentioning

confidence: 99%

Beyond Capture: Circulating Tumor Cell Release and Single‐Cell Analysis

Sharma

et al. 2019

Small Methods

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As the most promising “liquid biopsy”, analysis of circulating tumor cells (CTCs) provides noninvasive access to obtain biological information of solid tumors. It not only advances the understanding of the mechanisms underlying tumor metastasis, relapse, and chemoresistance, but also promotes the development of precision medicine. Over the past decade, the rapid advances in micro/nanomaterials and microfluidics have overcome the technical challenges of capturing CTCs, allowing efficient enrichment and sensitive detection. Beyond the capture of CTCs, major challenges of in‐depth CTC analysis are the release of CTCs from capture platforms and the inherent heterogeneity in CTCs. A variety of technologies have now emerged for release and single‐cell analysis of CTCs. This review focuses on recent progress along this direction. First, different release strategies are outlined and discussed, including their design principles and characteristics (merits and limitations). Then, existing methods for single CTC analysis at the genomic, transcriptomic, proteomic, and functional level are summarized. Finally, some perspectives are provided on current development trends, future research directions, and challenges of CTC studies.

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Next-generation Sequencing (NGS) Analysis on Single Circulating Tumor Cells (CTCs) with No Need of Whole-genome Amplification (WGA)

Cited by 33 publications

References 32 publications

DEPArray™ system: An automatic image‐based sorter for isolation of pure circulating tumor cells

DEPArray™ system: An automatic image‐based sorter for isolation of pure circulating tumor cells

Liquid biopsy - emergence of a new era in personalized cancer care

Beyond Capture: Circulating Tumor Cell Release and Single‐Cell Analysis

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