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Cancers
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Liquid Biopsy: Latest Advances and Future Challenges
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Liquid Biopsy: Latest Advances and Future Challenges

A special issue of Cancers (ISSN 2072-6694). This special issue belongs to the section "Molecular Cancer Biology".

Deadline for manuscript submissions: closed (31 March 2020) | Viewed by 102224

Special Issue Editors

Prof. Dr. Evi S. Lianidou

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Guest Editor
Analysis of Circulating Tumor Cells Lab, Department of Chemistry, University of Athens, University Campus, 15771 Athens, Greece
Interests: liquid biopsy; circulating tumor cells (CTCs); circulating tumor DNA (ctDNA); circulating miRNAs; DNA methylation; gene expression; molecular assays
Special Issues, Collections and Topics in MDPI journals
Prof. Dr. Klaus Pantel

E-Mail Website
Guest Editor
Department of Tumor Biology, University Medical Center Hamburg-Eppendorf, 20251 Hamburg, Germany
Dr. Catherine Alix-Panabieres

E-Mail Website
Guest Editor
University Medical Center of Montpellier, IURC, Laboratory of Rare Human Circulating Cells (LCCRH), 641 avenue du Doyen Gaston Giraud, 34093 Montpellier, France
Interests: circulating tumor cells (CTCs); liquid biopsy; biomarkers, metastasis-competent CTCs
Special Issues, Collections and Topics in MDPI journals

Special Issue Information

Dear Colleagues,

This Special Issue of Cancers will consist of selected papers from The 4th ACTC (Advances in Circulating Tumor Cells) “Liquid Biopsy: Latest Advances and Future Challenges” meeting that took place in Corfu (Greece), on 2–5 October 2019.

The 4th ACTC meeting was highly interactive, stimulated intensive discussions, and was attended by 288 participants from 29 countries. Networking and knowledge sharing across basic researchers, clinicians, and company representatives was an important part of this event. The meeting brought together researchers from academia, clinicians, and technology providers that presented, discussed, and shared the latest information on liquid biopsy in a nice and relaxing atmosphere.

A critical mass of world-recognized pioneer researchers and clinical oncologists that have major contributions in the field committed to the 4th ACTC meeting through state-of-the-art presentations, stimulating discussions, and sharing a lot of unpublished data. A highly interested critical mass of participants from all over the world had the chance to interact, establish collaborations, and present their latest data in a unique, relaxing, and stimulating environment.

State of the art research on the latest technical advancements in the isolation and molecular characterization of CTCs, ctDNA analysis, miRNAs, and exosomes was presented through 34 plenary lectures, 27 selected oral presentations, and 108 posters. There were also 10 high-level industry-sponsored presentations on the latest advances in liquid biopsy technologies, and all sessions were highly attended. The latest findings on the clinical trials based on the liquid biopsy approach were thoroughly discussed. Three travel grants and three poster awards were granted to the best submitted abstracts.

This Special Issue in Cancers (IF: 6.162) will publish a selection of papers presented at the 4th ACTC meeting. The Issue will focus on state of the art analytical developments, on the latest findings on CTCs and ctDNA analysis, and clinical evidence regarding the implementation of liquid biopsy in clinical practice.

Prof. Dr. Evi S. Lianidou
Prof. Dr. Klaus Pantel
Dr. Catherine Alix-Panabières
Guest Editors

Manuscript Submission Information

Manuscripts should be submitted online at www.mdpi.com by registering and logging in to this website. Once you are registered, click here to go to the submission form. Manuscripts can be submitted until the deadline. All submissions that pass pre-check are peer-reviewed. Accepted papers will be published continuously in the journal (as soon as accepted) and will be listed together on the special issue website. Research articles, review articles as well as short communications are invited. For planned papers, a title and short abstract (about 100 words) can be sent to the Editorial Office for announcement on this website.

Submitted manuscripts should not have been published previously, nor be under consideration for publication elsewhere (except conference proceedings papers). All manuscripts are thoroughly refereed through a single-blind peer-review process. A guide for authors and other relevant information for submission of manuscripts is available on the Instructions for Authors page. Cancers is an international peer-reviewed open access semimonthly journal published by MDPI.

Please visit the Instructions for Authors page before submitting a manuscript. The Article Processing Charge (APC) for publication in this open access journal is 2900 CHF (Swiss Francs). Submitted papers should be well formatted and use good English. Authors may use MDPI's English editing service prior to publication or during author revisions.

Keywords

  • liquid biopsy
  • circulating tumor cells
  • circulating tumor DNA
  • CTCs
  • ctDNA
  • circulating miRNAs
  • exosomes
  • extracellular vesicles
  • precision medicine

Published Papers (23 papers)

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28 pages, 4743 KiB  
Article
Diversity of Epithelial-Mesenchymal Phenotypes in Circulating Tumour Cells from Prostate Cancer Patient-Derived Xenograft Models
by Sara Hassan, Tony Blick, Erik W. Thompson and Elizabeth D. Williams
Cancers 2021, 13(11), 2750; https://doi.org/10.3390/cancers13112750 - 01 Jun 2021
Cited by 19 | Viewed by 3755
Abstract
Metastasis is the leading cause of cancer-related deaths worldwide. The epithelial-mesenchymal plasticity (EMP) status of primary tumours has relevance to metastatic potential and therapy resistance. Circulating tumour cells (CTCs) provide a window into the metastatic process, and molecular characterisation of CTCs in comparison [...] Read more.
Metastasis is the leading cause of cancer-related deaths worldwide. The epithelial-mesenchymal plasticity (EMP) status of primary tumours has relevance to metastatic potential and therapy resistance. Circulating tumour cells (CTCs) provide a window into the metastatic process, and molecular characterisation of CTCs in comparison to their primary tumours could lead to a better understanding of the mechanisms involved in the metastatic cascade. In this study, paired blood and tumour samples were collected from four prostate cancer patient-derived xenograft (PDX) models (BM18, LuCaP70, LuCaP96, LuCaP105) and assessed using an EMP-focused, 42 gene human-specific, nested quantitative RT-PCR assay. CTC burden varied amongst the various xenograft models with LuCaP96 having the highest number of CTCs per mouse (mean: 704; median: 31) followed by BM18 (mean: 101; median: 21), LuCaP70 (mean: 73; median: 16) and LuCaP105 (mean: 57; median: 6). A significant relationship was observed between tumour size and CTC number (p = 0.0058). Decreased levels of kallikrein-related peptidase 3 (KLK3) mRNA (which encodes prostate-specific antigen; PSA) were observed in CTC samples from all four models compared to their primary tumours. Both epithelial- and mesenchymal-associated genes were commonly expressed at higher levels in CTCs compared to the bulk primary tumour, although some common EMT-associated genes (CDH1, VIM, EGFR, EPCAM) remained unchanged. Immunofluorescence co-staining for pan-cytokeratin (KRT) and vimentin (VIM) indicated variable proportions of CTCs across the full EMP axis, even in the same model. EMP hybrids predominated in the BM18 and LuCaP96 models, but were not detected in the LuCaP105 model, and variable numbers of KRT+ and human VIM+ cells were observed in each model. SERPINE1, which encodes plasminogen activator inhibitor-1 (PAI-1), was enriched at the RNA level in CTCs compared to primary tumours and was the most commonly expressed mesenchymal gene in the CTCs. Co-staining for SERPINE1 and KRT revealed SERPINE1+ cells in 7/11 samples, six of which had SERPINE+KRT+ CTCs. Cell size variation was observed in CTCs. The majority of samples (8/11) contained larger CTCs ranging from 15.3 to 37.8 µm, whilst smaller cells (10.7 ± 4.1 µm, similar in size to peripheral blood mononuclear cells (PBMCs)) were identified in 6 of 11 samples. CTC clusters were also identified in 9/11 samples, containing 2–100 CTCs per cluster. Where CTC heterogeneity was observed in the clusters, epithelial-like cells (KRT+VIM−) were located on the periphery of the cluster, forming a layer around hybrid (KRT+VIM+) or mesenchymal-like (KRT−VIM+) cells. The CTC heterogeneity observed in these models emphasises the complexity in CTC isolation and classification and supports the increasingly recognised importance of the epithelial-mesenchymal hybrid state in cancer progression and metastasis. Full article
(This article belongs to the Special Issue Liquid Biopsy: Latest Advances and Future Challenges)
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27 pages, 3629 KiB  
Article
Cysteine-Rich Angiogenic Inducer 61: Pro-Survival Function and Role as a Biomarker for Disseminating Breast Cancer Cells
by Kai Bartkowiak, Isabel Heidrich, Marcel Kwiatkowski, Tobias M. Gorges, Antje Andreas, Maria Geffken, Karl Verpoort, Volkmar Müller, Hartmut Schlüter and Klaus Pantel
Cancers 2021, 13(3), 563; https://doi.org/10.3390/cancers13030563 - 02 Feb 2021
Cited by 6 | Viewed by 2905
Abstract
(1) Background: the early detection of cancer cells in the blood or bone marrow of breast cancer patients improves the understanding of metastasis. Disseminating tumor cells in the bone marrow with a pronounced manifestation of mesenchymal markers (mDTC) are difficult to detect by [...] Read more.
(1) Background: the early detection of cancer cells in the blood or bone marrow of breast cancer patients improves the understanding of metastasis. Disseminating tumor cells in the bone marrow with a pronounced manifestation of mesenchymal markers (mDTC) are difficult to detect by epithelial markers, but they are relevant in the initiation of metastasis. (2) Methods: the breast cancer mDTC cell line BC-M1 was analyzed by mass spectrometry, which revealed high levels of the protein-cysteine–rich angiogenic inducer 61 (Cyr61). The function of Cyr61 was investigated using shRNA and hypoxia. Peripheral blood samples from 35 breast cancer patients were investigated for CTCs defined as cytokeratin-positive/CD45-negative cells. (3) Results: the Cyr61 levels are elevated in mDTC lines from breast, lung, and prostate cancer patients. The loss of Cyr61 resulted in the diminished expression of hypoxia-inducible factor 1-alpha, and increased apoptosis. Cyr61 was present in 47 (43%) of the 109 detected circulating tumor cells (CTCs), while the blood and bone marrow cells from healthy controls were Cyr61-negative. (4) Conclusions: Cyr61 is expressed in mDTC lines, supports the viability of cancer cells, and classifies a new subset of cytokeratin-positive CTCs, which deserves further investigation. Full article
(This article belongs to the Special Issue Liquid Biopsy: Latest Advances and Future Challenges)
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24 pages, 5104 KiB  
Article
Integrated Assessment of Circulating Cell-Free MicroRNA Signatures in Plasma of Patients with Melanoma Brain Metastasis
by Matias A. Bustos, Kevin D. Tran, Negin Rahimzadeh, Rebecca Gross, Selena Y. Lin, Yoshiaki Shoji, Tomohiro Murakami, Christine L. Boley, Linh T. Tran, Hunter Cole, Daniel F. Kelly, Steven O’Day and Dave S. B. Hoon
Cancers 2020, 12(6), 1692; https://doi.org/10.3390/cancers12061692 - 25 Jun 2020
Cited by 18 | Viewed by 3516
Abstract
Primary cutaneous melanoma frequently metastasizes to distant organs including the brain. Identification of cell-free microRNAs (cfmiRs) found in the blood can be used as potential body fluid biomarkers for detecting and monitoring patients with melanoma brain metastasis (MBM). In this pilot study, we [...] Read more.
Primary cutaneous melanoma frequently metastasizes to distant organs including the brain. Identification of cell-free microRNAs (cfmiRs) found in the blood can be used as potential body fluid biomarkers for detecting and monitoring patients with melanoma brain metastasis (MBM). In this pilot study, we initially aimed to identify cfmiRs in the blood of MBM patients. Normal donors plasma (healthy, n = 48) and pre-operative MBM patients’ plasma samples (n = 36) were compared for differences in >2000 microRNAs (miRs) using a next generation sequencing (NGS) probe-based assay. A 74 cfmiR signature was identified in an initial cohort of MBM plasma samples and then verified in a second cohort of MBM plasma samples (n = 24). Of these, only 58 cfmiRs were also detected in MBM tissues (n = 24). CfmiR signatures were also found in patients who have lung and breast cancer brain metastasis (n = 13) and glioblastomas (n = 36) compared to MBM plasma samples. The 74 cfmiR signature and the latter cfmiR signatures were then compared. We found a 6 cfmiR signature that was commonly upregulated in MBM plasma samples in all of the comparisons, and a 29 cfmiR signature that distinguishes MBM patients from normal donors’ samples. In addition, we assessed for cfmiRs in plasma (n = 20) and urine (n = 14) samples collected from metastatic melanoma patients receiving checkpoint inhibitor immunotherapy (CII). Pre- and post-treatment samples showed consistent changes in cfmiRs. Analysis of pre- and post-treatment plasma samples showed 8 differentially expressed (DE) cfmiRs that overlapped with the 35 cfmiR signature found in MBM patients. In paired pre-treatment plasma and urine samples receiving CII 8 cfmiRs overlapped. This study identified specific cfmiRs in MBM plasma samples that may potentially allow for assessment of melanoma patients developing MBM. The cfmiR signatures identified in both blood and urine may have potential utility to assess CII responses after further validation. Full article
(This article belongs to the Special Issue Liquid Biopsy: Latest Advances and Future Challenges)
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19 pages, 4897 KiB  
Article
Molecular Interplay between Dormant Bone Marrow-Resident Cells (BMRCs) and CTCs in Breast Cancer
by Debasish Boral, Haowen N. Liu, S. Ray Kenney and Dario Marchetti
Cancers 2020, 12(6), 1626; https://doi.org/10.3390/cancers12061626 - 19 Jun 2020
Cited by 8 | Viewed by 4127
Abstract
Despite widespread knowledge that bone marrow-resident breast cancer cells (BMRCs) affect tumor progression, signaling mechanisms of BMRCs implicated in maintaining long-term dormancy have not been characterized. To overcome these hurdles, we developed a new experimental model of clinical dormancy employing patient-isolated Circulating Tumor [...] Read more.
Despite widespread knowledge that bone marrow-resident breast cancer cells (BMRCs) affect tumor progression, signaling mechanisms of BMRCs implicated in maintaining long-term dormancy have not been characterized. To overcome these hurdles, we developed a new experimental model of clinical dormancy employing patient-isolated Circulating Tumor Cells (de novo CTCs) and their injection in xenografts with subsequent tumor monitoring and CTC characterization (ex vivo CTCs). We hypothesized that significant distinctions exist between signaling pathways of bone marrow-homing vs metastasis-competent CTCs upon transplantation in xenografts. Comparative transcriptomic analyses of ex vivo vs de novo CTCs identified increased mTOR signaling—a critical pathway frequently dysregulated in breast cancer and implicated in cell survival and dormancy—with contrasting actions by its two complementary arms (mTORC2/mTORC1). Heightened mTORC2 downstream targets augmented quiescent CTCs (Ki67−/RBL2+ cells) in paired breast cancer tissues, along with high mTORC2 activity in solitary BMRCs and tissue-resident CTCs. Further, shRNA mediated the knockdown of RICTOR, an essential component of mTORC2, and augmented Ki67/PCNA biomarker expression and proliferation. Collectively, these findings suggest that the balance between mTORC1 vs mTORC2 signaling regulates CTC-associated mitotic and/or dormancy characteristics. Full article
(This article belongs to the Special Issue Liquid Biopsy: Latest Advances and Future Challenges)
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16 pages, 2389 KiB  
Article
Technical Evaluation of Commercial Mutation Analysis Platforms and Reference Materials for Liquid Biopsy Profiling
by Sabrina Weber, Benjamin Spiegl, Samantha O. Perakis, Christine M. Ulz, Peter M. Abuja, Karl Kashofer, Paul van der Leest, Maria Aguirre Azpurua, Menno Tamminga, Dan Brudzewsky, Dominic G. Rothwell, Sumitra Mohan, Alexander Sartori, Rita Lampignano, Yves Konigshofer, Markus Sprenger-Haussels, Harriet Wikman, Inger R. Bergheim, Vera Kloten, Ed Schuuring, Michael R. Speicher and Ellen Heitzeradd Show full author list remove Hide full author list
Cancers 2020, 12(6), 1588; https://doi.org/10.3390/cancers12061588 - 16 Jun 2020
Cited by 44 | Viewed by 10963
Abstract
Molecular profiling from liquid biopsy, in particular cell-free DNA (cfDNA), represents an attractive alternative to tissue biopsies for the detection of actionable targets and tumor monitoring. In addition to PCR-based assays, Next Generation Sequencing (NGS)-based cfDNA assays are now commercially available and are [...] Read more.
Molecular profiling from liquid biopsy, in particular cell-free DNA (cfDNA), represents an attractive alternative to tissue biopsies for the detection of actionable targets and tumor monitoring. In addition to PCR-based assays, Next Generation Sequencing (NGS)-based cfDNA assays are now commercially available and are being increasingly adopted in clinical practice. However, the validity of these products as well as the clinical utility of cfDNA in the management of patients with cancer has yet to be proven. Within framework of the Innovative Medicines Initiative (IMI) program CANCER-ID we evaluated the use of commercially available reference materials designed for ctDNA testing and cfDNA derived from Diagnostic Leukaphereses (DLA) for inter- and intra-assay as well as intra- and inter-laboratory comparisons. In three experimental setups, a broad range of assays including ddPCR, MassARRAY and various NGS-based assays were tested. We demonstrate that both reference materials with predetermined VAFs and DLA samples are extremely useful for the performance assessment of mutation analysis platforms. Moreover, our data indicate a substantial variability of NGS assays with respect to sensitivity and specificity highlighting the importance of extensive validation of the test performance before offering these tests in clinical routine practice. Full article
(This article belongs to the Special Issue Liquid Biopsy: Latest Advances and Future Challenges)
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26 pages, 3828 KiB  
Article
Optimization of the Enrichment of Circulating Tumor Cells for Downstream Phenotypic Analysis in Patients with Non-Small Cell Lung Cancer Treated with Anti-PD-1 Immunotherapy
by Maria A Papadaki, Afroditi I Sotiriou, Christina Vasilopoulou, Maria Filika, Despoina Aggouraki, Panormitis G Tsoulfas, Christina A Apostolopoulou, Konstantinos Rounis, Dimitrios Mavroudis and Sofia Agelaki
Cancers 2020, 12(6), 1556; https://doi.org/10.3390/cancers12061556 - 12 Jun 2020
Cited by 26 | Viewed by 3615
Abstract
The current study aimed at the optimization of circulating tumor cell (CTC) enrichment for downstream protein expression analyses in non-small cell lung cancer (NSCLC) to serve as a tool for the investigation of immune checkpoints in real time. Different enrichment approaches—ficoll density, erythrolysis, [...] Read more.
The current study aimed at the optimization of circulating tumor cell (CTC) enrichment for downstream protein expression analyses in non-small cell lung cancer (NSCLC) to serve as a tool for the investigation of immune checkpoints in real time. Different enrichment approaches—ficoll density, erythrolysis, their combination with magnetic separation, ISET, and Parsortix—were compared in spiking experiments using the A549, H1975, and SKMES-1 NSCLC cell lines. The most efficient methods were tested in patients (n = 15) receiving immunotherapy targeting programmed cell death-1 (PD-1). Samples were immunofluorescently stained for a) cytokeratins (CK)/epithelial cell adhesion molecule (EpCAM)/leukocyte common antigen (CD45), and b) CK/programmed cell death ligand-1 (PD-L1)/ indoleamine-2,3-dioxygenase (IDO). Ficoll, ISET, and Parsortix presented the highest yields and compatibility with phenotypic analysis; however, at the patient level, they provided discordant CTC positivity (13%, 33%, and 60% of patients, respectively) and enriched for distinct CTC populations. IDO and PD-L1 were expressed in 44% and 33% and co-expressed in 19% of CTCs. CTC detection was associated with progressive disease (PD) (p = 0.006), reduced progression-free survival PFS (p = 0.007), and increased risk of relapse (hazard ratio; HR: 10.733; p = 0.026). IDO-positive CTCs were associated with shorter PFS (p = 0.039) and overall survival OS (p = 0.021) and increased risk of death (HR: 5.462; p = 0.039). The current study indicates that CTC analysis according to distinct immune checkpoints is feasible and may provide valuable biomarkers to monitor NSCLC patients treated with anti-PD-1 agents. Full article
(This article belongs to the Special Issue Liquid Biopsy: Latest Advances and Future Challenges)
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9 pages, 860 KiB  
Article
RNA-Sequencing of Tumor-Educated Platelets, a Novel Biomarker for Blood-Based Sarcoma Diagnostics
by Kimberley M. Heinhuis, Sjors G. J. G. In ’t Veld, Govert Dwarshuis, Daan van den Broek, Nik Sol, Myron G. Best, Frits van Coevorden, Rick L. Haas, Jos H. Beijnen, Winan J. van Houdt, Tom Würdinger and Neeltje Steeghs
Cancers 2020, 12(6), 1372; https://doi.org/10.3390/cancers12061372 - 27 May 2020
Cited by 33 | Viewed by 3496
Abstract
Sarcoma is a heterogeneous group of rare malignancies arising from mesenchymal tissues. Recurrence rates are high and methods for early detection by blood-based biomarkers do not exist. Hence, development of blood-based liquid biopsies as disease recurrence monitoring biomarkers would be an important step [...] Read more.
Sarcoma is a heterogeneous group of rare malignancies arising from mesenchymal tissues. Recurrence rates are high and methods for early detection by blood-based biomarkers do not exist. Hence, development of blood-based liquid biopsies as disease recurrence monitoring biomarkers would be an important step forward. Recently, it has been shown that tumor-educated platelets (TEPs) harbor specific spliced ribonucleic acid(RNA)-profiles. These RNA-repertoires are potentially applicable for cancer diagnostics. We aim to evaluate the potential of TEPs for blood-based diagnostics of sarcoma patients. Fifty-seven sarcoma patients (active disease), 38 former sarcoma patients (cancer free for ≥3 years) and 65 healthy donors were included. RNA was isolated from platelets and sequenced. Quantified read counts were processed with self-learning particle-swarm optimization-enhanced thromboSeq analysis and subjected to analysis of variance (ANOVA) statistics. Highly correlating spliced platelet messenger RNAs (mRNAs) of sarcoma patients were compared to controls (former sarcoma + healthy donors) to identify a quantitative sarcoma-specific signature measure, the TEP-score. ANOVA analysis identified distinctive platelet RNA expression patterns of 2647 genes (false discovery rate <0.05) in sarcoma patients as compared to controls. The self-learning algorithm reached a diagnostic accuracy of 87% (validation set only; n = 53 samples, area under the curve (AUC): 0.93, 95% confidence interval (CI): 0.86–1). Our data indicates that TEP RNA-based liquid biopsies may enable for sarcoma diagnostics. Full article
(This article belongs to the Special Issue Liquid Biopsy: Latest Advances and Future Challenges)
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15 pages, 3520 KiB  
Article
Tailoring Chemometric Models on Blood-Derived Cultures Secretome to Assess Personalized Cancer Risk Score
by Maria Laura Coluccio, Francesco Gentile, Ivan Presta, Giuseppe Donato, Nicola Coppedè, Immanuel Valprapuram, Chiara Mignogna, Annamaria Lavecchia, Federica Figuccia, Virginia M. Garo, Enzo Di Fabrizio, Patrizio Candeloro, Giuseppe Viglietto and Natalia Malara
Cancers 2020, 12(6), 1362; https://doi.org/10.3390/cancers12061362 - 26 May 2020
Cited by 7 | Viewed by 2416
Abstract
The molecular protonation profiles obtained by means of an organic electrochemical transistor, which is used for analysis of molecular products released by blood-derived cultures, contain a large amount of information The transistor is based on the conductive polymer PEDOT:PSS comprising super hydrophobic SU8 [...] Read more.
The molecular protonation profiles obtained by means of an organic electrochemical transistor, which is used for analysis of molecular products released by blood-derived cultures, contain a large amount of information The transistor is based on the conductive polymer PEDOT:PSS comprising super hydrophobic SU8 pillars positioned on the substrate to form a non-periodic square lattice to measure the state of protonation on secretomes derived from liquid biopsies. In the extracellular space of cultured cells, the number of glycation products increase, driven both by a glycolysis metabolism and by a compromised function of the glutathione redox system. Glycation products are a consequence of the interaction of the reactive aldehydes and side glycolytic products with other molecules. As a result, the amount of the glycation products reflects the anti-oxidative cellular reserves, counteracting the reactive aldehyde production of which both the secretome protonation profile and cancer risk are related. The protonation profiles can be profitably exploited through the use of mathematical techniques and multivariate statistics. This study provides a novel chemometric approach for molecular analysis of protonation and discusses the possibility of constructing a predictive cancer risk model based on the exploration of data collected by conventional analysis techniques and novel nanotechnological devices. Full article
(This article belongs to the Special Issue Liquid Biopsy: Latest Advances and Future Challenges)
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18 pages, 1046 KiB  
Article
Liquid Biopsy Serial Monitoring of Treatment Responses and Relapse in Advanced Esophageal Squamous Cell Carcinoma
by Josephine Mun Yee Ko, Hoi Yan Ng, Ka On Lam, Keith Wan Hang Chiu, Dora Lai Wan Kwong, Anthony Wing Ip Lo, Jean Chrysei Wong, Roger Chia Wei Lin, Henry Chun Hung Fong, Jason Ying Ki Li, Wei Dai, Simon Law and Maria Li Lung
Cancers 2020, 12(6), 1352; https://doi.org/10.3390/cancers12061352 - 26 May 2020
Cited by 14 | Viewed by 2775
Abstract
(1) Background: Early predictive markers to track treatment responses are needed for advanced esophageal squamous cell carcinoma (ESCC) patients. We examined the prognostication and risk stratification role of liquid biopsy serial monitoring for this deadly cancer. (2) Methods: Circulating tumor cells (CTCs) and [...] Read more.
(1) Background: Early predictive markers to track treatment responses are needed for advanced esophageal squamous cell carcinoma (ESCC) patients. We examined the prognostication and risk stratification role of liquid biopsy serial monitoring for this deadly cancer. (2) Methods: Circulating tumor cells (CTCs) and plasma cell-free DNA (cfDNA) were isolated from 60 ESCC patients treated by chemotherapy (CT) at five serial timepoints: baseline (CTC1/cfDNA1), CT pre-cycle III (CTC2/cfDNA2), CT post-cycle IV, end of CT and relapse. (3) Results: In 45/57 ESCC patients with evaluable CTC counts at CT pre-cycle III, positive CTC2 (≥3 CTCs) is independently associated with response at interim reassessment and progression-free survival (PFS) in multivariate analysis. In 42/57 ESCC patients with changes of CTC1/CTC2 and cfDNA1/cfDNA2, patients categorized into four risk groups based on the number of favorable and unfavorable changes of CTC1/CTC2 and cfDNA1/cfDNA2, were independently associated with overall survival (OS) by multivariate analysis. (4) Conclusions: CTC counts at pre-cycle III are independently associated with response at interim reassessment and PFS. Combined changes of CTC counts and cfDNA levels from baseline to pre-cycle III are independently associated with OS. Longitudinal liquid biopsy serial monitoring provides complementary information for prediction and prognosis for CT responses in advanced ESCC. Full article
(This article belongs to the Special Issue Liquid Biopsy: Latest Advances and Future Challenges)
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14 pages, 2034 KiB  
Article
Comparison of Circulating Cell-Free DNA Extraction Methods for Downstream Analysis in Cancer Patients
by Paul van der Leest, Pieter A. Boonstra, Arja ter Elst, Léon C. van Kempen, Marco Tibbesma, Jill Koopmans, Anneke Miedema, Menno Tamminga, Harry J. M. Groen, Anna K. L. Reyners and Ed Schuuring
Cancers 2020, 12(5), 1222; https://doi.org/10.3390/cancers12051222 - 13 May 2020
Cited by 36 | Viewed by 5358
Abstract
Circulating cell-free DNA (ccfDNA) may contain DNA originating from the tumor in plasma of cancer patients (ctDNA) and enables noninvasive cancer diagnosis, treatment predictive testing, and response monitoring. A recent multicenter evaluation of workflows by the CANCER-ID consortium using artificial spiked-in plasma showed [...] Read more.
Circulating cell-free DNA (ccfDNA) may contain DNA originating from the tumor in plasma of cancer patients (ctDNA) and enables noninvasive cancer diagnosis, treatment predictive testing, and response monitoring. A recent multicenter evaluation of workflows by the CANCER-ID consortium using artificial spiked-in plasma showed significant differences and consequently the importance of carefully selecting ccfDNA extraction methods. Here, the quantity and integrity of extracted ccfDNA from the plasma of cancer patients were assessed. Twenty-one cancer patient-derived cell-free plasma samples were selected to compare the Qiagen CNA, Maxwell RSC ccfDNA plasma, and Zymo manual quick ccfDNA kit. High-volume citrate plasma samples collected by diagnostic leukapheresis from six cancer patients were used to compare the Qiagen CNA (2 mL) and QIAamp MinElute ccfDNA kit (8 mL). This study revealed similar integrity and similar levels of amplified short-sized fragments and tumor-specific mutants comparing the CNA and RSC kits. However, the CNA kit consistently showed the highest yield of ccfDNA and short-sized fragments, while the RSC and ME kits showed higher variant allelic frequencies (VAFs). Our study pinpoints the importance of standardizing preanalytical conditions as well as consensus on defining the input of ccfDNA to accurately detect ctDNA and be able to compare results in a clinical routine practice, within and between clinical studies. Full article
(This article belongs to the Special Issue Liquid Biopsy: Latest Advances and Future Challenges)
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14 pages, 1362 KiB  
Article
PIM-1 Is Overexpressed at a High Frequency in Circulating Tumor Cells from Metastatic Castration-Resistant Prostate Cancer Patients
by Athina Markou, Eleni Tzanikou, Areti Strati, Martha Zavridou, Sophia Mastoraki, Evangelos Bournakis and Evi Lianidou
Cancers 2020, 12(5), 1188; https://doi.org/10.3390/cancers12051188 - 08 May 2020
Cited by 12 | Viewed by 3729
Abstract
PIM-1 is an oncogene involved in cell cycle progression, cell growth, cell survival and therapy resistance, activated in many types of cancer, and is now considered as a very promising target for cancer therapy. We report for the first time that PIM-1 is [...] Read more.
PIM-1 is an oncogene involved in cell cycle progression, cell growth, cell survival and therapy resistance, activated in many types of cancer, and is now considered as a very promising target for cancer therapy. We report for the first time that PIM-1 is overexpressed in circulating tumor cells (CTCs) from metastatic castration-resistant prostate cancer patients (mCRPC). We first developed and validated a highly sensitive RT-qPCR assay for quantification of PIM-1 transcripts. We further applied this assay to study PIM-1 expression in EpCAM(+) CTC fraction isolated from 64 peripheral blood samples of 50 mCRPC patients. CTC enumeration in all samples was performed using the FDA-cleared CellSearch® system. PIM-1 overexpression was detected in 24/64 (37.5%) cases, while in 20/24 (83.3%) cases that were positive for PIM-1 expression, at least one CTC/7.5 mL PB was detected in the CellSearch®. Our data indicate that PIM-1 overexpression is observed at high frequency in CTCs from mCRPC patients and this finding, in combination with androgen receptor splice variant 7 (AR-V7) expression in CTCs, suggest its potential role as a very promising target for cancer therapy. We strongly believe that PIM-1 overexpression in EpCAM(+) CTC fraction merits to be further evaluated and validated as a non-invasive circulating tumor biomarker in a large and well-defined patient cohort with mCRPC. Full article
(This article belongs to the Special Issue Liquid Biopsy: Latest Advances and Future Challenges)
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12 pages, 6123 KiB  
Article
Multicenter Evaluation of Independent High-Throughput and RT-qPCR Technologies for the Development of Analytical Workflows for Circulating miRNA Analysis
by Anna Babayan, Martin H. D. Neumann, Andrei Herdean, Jonathan M. Shaffer, Melanie Janning, Franca Kobus, Sonja Loges, Francesca Di Pasquale, Mikael Kubista, Martin Schlumpberger, Rita Lampignano, Thomas Krahn, Thomas Schlange, Markus Sprenger-Haussels, Klaus Pantel and Vera Kloten
Cancers 2020, 12(5), 1166; https://doi.org/10.3390/cancers12051166 - 05 May 2020
Cited by 11 | Viewed by 3448
Abstract
Background: Among emerging circulating biomarkers, miRNA has the potential to detect lung cancer and follow the course of the disease. However, miRNA analysis deserves further standardization before implementation into clinical trials or practice. Here, we performed international ring experiments to explore (pre)-analytical factors [...] Read more.
Background: Among emerging circulating biomarkers, miRNA has the potential to detect lung cancer and follow the course of the disease. However, miRNA analysis deserves further standardization before implementation into clinical trials or practice. Here, we performed international ring experiments to explore (pre)-analytical factors relevant to the outcome of miRNA blood tests in the context of the EU network CANCER-ID. Methods: Cell-free (cfmiRNA) and extracellular vesicle-derived miRNA (EVmiRNA) were extracted using the miRNeasy Serum/Plasma Advanced, and the ExoRNeasy Maxi kit, respectively, in a plasma cohort of 27 NSCLC patients and 20 healthy individuals. Extracted miRNA was investigated using small RNA sequencing and hybridization platforms. Validation of the identified miRNA candidates was performed using quantitative PCR. Results: We demonstrate the highest read counts in healthy individuals and NSCLC patients using QIAseq. Moreover, QIAseq showed 15.9% and 162.9% more cfmiRNA and EVmiRNA miRNA counts, respectively, in NSCLC patients compared to healthy control samples. However, a systematic comparison of selected miRNAs revealed little agreement between high-throughput platforms, thus some miRNAs are detected with one technology, but not with the other. Adding to this, 35% (9 of 26) of selected miRNAs in the cfmiRNA and 42% (11 of 26) in the EVmiRNA fraction were differentially expressed by at least one qPCR platform; about half of the miRNAs (54%) were concordant for both platforms. Conclusions: Changing of (pre)-analytical methods of miRNA analysis has a significant impact on blood test results and is therefore a major confounding factor. In addition, to confirm miRNA biomarker candidates screening studies should be followed by targeted validation using an independent platform or technology. Full article
(This article belongs to the Special Issue Liquid Biopsy: Latest Advances and Future Challenges)
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15 pages, 1820 KiB  
Article
Inflammation-Based Scores Increase the Prognostic Value of Circulating Tumor Cells in Primary Breast Cancer
by Svetlana Miklikova, Gabriel Minarik, Tatiana Sedlackova, Jana Plava, Marina Cihova, Silvia Jurisova, Katarina Kalavska, Marian Karaba, Juraj Benca, Bozena Smolkova and Michal Mego
Cancers 2020, 12(5), 1134; https://doi.org/10.3390/cancers12051134 - 01 May 2020
Cited by 23 | Viewed by 3445
Abstract
A correlation between circulating tumor cells (CTCs) and monocytes in metastatic breast cancer (BC), where CTCs and monocyte-to-lymphocyte ratio (MLR) were predictors of overall survival (OS), was recently shown. Herein, we aimed to assess the association between CTCs and the complete blood count [...] Read more.
A correlation between circulating tumor cells (CTCs) and monocytes in metastatic breast cancer (BC), where CTCs and monocyte-to-lymphocyte ratio (MLR) were predictors of overall survival (OS), was recently shown. Herein, we aimed to assess the association between CTCs and the complete blood count (CBC)-derived inflammation-based scores in 284 primary BC patients. CTCs were determined in CD45-depleted peripheral blood mononuclear cells by real time-PCR. This method allowed us to detect a subset of CTCs with an epithelial-to-mesenchymal transition phenotype (CTC EMT), previously associated with inferior outcomes in primary BC. In the present study, CTC EMT positivity (hazard ratio (HR) = 2.4; 95% CI 1.20–4.66, p = 0.013) and elevated neutrophil-to-lymphocyte ratio (NLR) (HR = 2.20; 95% CI 1.07–4.55; p = 0.033) were associated with shorter progression-free survival (PFS) in primary BC patients. Multivariate analysis showed that CTC EMT-positive patients with NLR ≥ 3 had 8.6 times increased risk of disease recurrence (95% CI 2.35–31.48, p = 0.001) compared with CTC EMT-negative patients with NLR < 3. Similarly, disease recurrence was 13.14 times more likely in CTC EMT-positive patients with MLR ≥ 0.34 (95% CI 4.35–39.67, p < 0.001). Given its low methodological and financial demands, the CBC-derived inflammation-based score determination could, after broader validation, significantly improve the prognostication of BC patients. Full article
(This article belongs to the Special Issue Liquid Biopsy: Latest Advances and Future Challenges)
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Graphical abstract

19 pages, 2853 KiB  
Article
Analysis of a Real-World Cohort of Metastatic Breast Cancer Patients Shows Circulating Tumor Cell Clusters (CTC-clusters) as Predictors of Patient Outcomes
by Clotilde Costa, Laura Muinelo-Romay, Victor Cebey-López, Thais Pereira-Veiga, Inés Martínez-Pena, Manuel Abreu, Alicia Abalo, Ramón M. Lago-Lestón, Carmen Abuín, Patricia Palacios, Juan Cueva, Roberto Piñeiro and Rafael López-López
Cancers 2020, 12(5), 1111; https://doi.org/10.3390/cancers12051111 - 29 Apr 2020
Cited by 38 | Viewed by 3977
Abstract
Circulating tumor cell (CTC) enumeration has emerged as a powerful biomarker for the assessment of prognosis and the response to treatment in metastatic breast cancer (MBC). Moreover, clinical evidences show that CTC-cluster counts add prognostic information to CTC enumeration, however, their significance is [...] Read more.
Circulating tumor cell (CTC) enumeration has emerged as a powerful biomarker for the assessment of prognosis and the response to treatment in metastatic breast cancer (MBC). Moreover, clinical evidences show that CTC-cluster counts add prognostic information to CTC enumeration, however, their significance is not well understood, and more clinical evidences are needed. We aim to evaluate the prognostic value of longitudinally collected single CTCs and CTC-clusters in a heterogeneous real-world cohort of 54 MBC patients. Blood samples were longitudinally collected at baseline and follow up. CTC and CTC-cluster enumeration was performed using the CellSearch® system. Associations with progression-free survival (PFS) and overall survival (OS) were evaluated using Cox proportional hazards modelling. Elevated CTC counts and CTC-clusters at baseline were significantly associated with a shorter survival time. In joint analysis, patients with high CTC counts and CTC-cluster at baseline were at a higher risk of progression and death, and longitudinal analysis showed that patients with CTC-clusters had significantly shorter survival compared to patients without clusters. Moreover, patients with CTC-cluster of a larger size were at a higher risk of death. A longitudinal analysis of a real-world cohort of MBC patients indicates that CTC-clusters analysis provides additional prognostic value to single CTC enumeration, and that CTC-cluster size correlates with patient outcome. Full article
(This article belongs to the Special Issue Liquid Biopsy: Latest Advances and Future Challenges)
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17 pages, 1871 KiB  
Article
Multimodal Targeted Deep Sequencing of Circulating Tumor Cells and Matched Cell-Free DNA Provides a More Comprehensive Tool to Identify Therapeutic Targets in Metastatic Breast Cancer Patients
by Corinna Keup, Markus Storbeck, Siegfried Hauch, Peter Hahn, Markus Sprenger-Haussels, Oliver Hoffmann, Rainer Kimmig and Sabine Kasimir-Bauer
Cancers 2020, 12(5), 1084; https://doi.org/10.3390/cancers12051084 - 27 Apr 2020
Cited by 16 | Viewed by 4040
Abstract
Cell-free DNA (cfDNA) and circulating tumor cells (CTCs) exhibit great potential for therapy management in oncology. We aimed to establish a multimodal liquid biopsy strategy that is usable with minimized blood volume to deconvolute the genomic complexity of metastatic breast cancer. CTCs were [...] Read more.
Cell-free DNA (cfDNA) and circulating tumor cells (CTCs) exhibit great potential for therapy management in oncology. We aimed to establish a multimodal liquid biopsy strategy that is usable with minimized blood volume to deconvolute the genomic complexity of metastatic breast cancer. CTCs were isolated from 10ml blood of 18 hormone receptor-positive and human epidermal growth factor receptor 2-negative (HER2-) metastatic breast cancer patients. cfDNA was isolated from plasma generated after CTC depletion and targeted sequencing analyses were conducted. PIK3CA and ESR1 variants were less common in CTC gDNA, while ERBB2 variants were only detected in CTC gDNA. A total of 62% of all cfDNA variants were recovered in the matched CTC gDNA, while 72% of all variants were unique in either cfDNA (14 variants) or CTC gDNA (104 variants). The percentage of patients with no detectable cfDNA variants or CTC gDNA variants was 17%/11%, but a combined analysis identified variants in 94% of all patients. In univariate and multivariate regression models, ESR1 variants in cfDNA and CTC gDNA correlated significantly with survival. We suggest a coordinated analysis of both fractions in order to provide a comprehensive genomic footprint that may contribute to identifying the most suitable therapy for each individual. Full article
(This article belongs to the Special Issue Liquid Biopsy: Latest Advances and Future Challenges)
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18 pages, 4225 KiB  
Article
Expansion of Circulating Tumor Cells from Patients with Locally Advanced Pancreatic Cancer Enable Patient Derived Xenografts and Functional Studies for Personalized Medicine
by Lianette Rivera-Báez, Ines Lohse, Eric Lin, Shreya Raghavan, Sarah Owen, Ramdane Harouaka, Kirk Herman, Geeta Mehta, Theodore S. Lawrence, Meredith A. Morgan, Kyle C. Cuneo and Sunitha Nagrath
Cancers 2020, 12(4), 1011; https://doi.org/10.3390/cancers12041011 - 20 Apr 2020
Cited by 27 | Viewed by 4880
Abstract
Improvement in pancreatic cancer treatment represents an urgent medical goal that has been hampered by the lack of predictive biomarkers. Circulating Tumor Cells (CTCs) may be able to overcome this issue by allowing the monitoring of therapeutic response and tumor aggressiveness through ex [...] Read more.
Improvement in pancreatic cancer treatment represents an urgent medical goal that has been hampered by the lack of predictive biomarkers. Circulating Tumor Cells (CTCs) may be able to overcome this issue by allowing the monitoring of therapeutic response and tumor aggressiveness through ex vivo expansion. The successful expansion of CTCs is challenging, due to their low numbers in blood and the high abundance of blood cells. Here, we explored the utility of pancreatic CTC cultures as a preclinical model for treatment response. CTCs were isolated from ten patients with locally advanced pancreatic cancer using the Labyrinth, a biomarker independent, size based, inertial microfluidic separation device. Three patient-derived CTC samples were successfully expanded in adherent and spheroid cultures. Molecular and functional characterization was performed on the expanded CTC lines. CTC lines exhibited KRAS mutations, consistent with pancreatic cancers. Additionally, we evaluated take rate and metastatic potential in vivo and examined the utility of CTC lines for cytotoxicity assays. Patient derived expanded CTCs successfully generated patient derived xenograft (PDX) models with a 100% take rate. Our results demonstrate that CTC cultures are possible and provide a valuable resource for translational pancreatic cancer research, while also providing meaningful insight into the development of distant metastasis, as well as treatment resistance. Full article
(This article belongs to the Special Issue Liquid Biopsy: Latest Advances and Future Challenges)
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17 pages, 2836 KiB  
Article
Magnetic-Based Enrichment of Rare Cells from High Concentrated Blood Samples
by Junhao Wu, Katharina Raba, Rosa Guglielmi, Bianca Behrens, Guus Van Dalum, Georg Flügen, Andreas Koch, Suraj Patel, Wolfram T. Knoefel, Nikolas H. Stoecklein and Rui P. L. Neves
Cancers 2020, 12(4), 933; https://doi.org/10.3390/cancers12040933 - 10 Apr 2020
Cited by 18 | Viewed by 3885
Abstract
Here, we tested two magnetic-bead based systems for the enrichment and detection of rare tumor cells in concentrated blood products. For that, the defined numbers of cells from three pancreatic cancer cell lines were spiked in 108 peripheral blood mononuclear cells (PBMNCs) [...] Read more.
Here, we tested two magnetic-bead based systems for the enrichment and detection of rare tumor cells in concentrated blood products. For that, the defined numbers of cells from three pancreatic cancer cell lines were spiked in 108 peripheral blood mononuclear cells (PBMNCs) concentrated in 1 mL, mimicking diagnostic leukapheresis (DLA) samples, and samples were processed for circulating tumor cells (CTC) enrichment with the IsoFlux or the KingFisher systems, using different types of magnetic beads from the respective technology providers. Beads were conjugated with different anti-EpCAM and MUC-1 antibodies. Recovered cells were enumerated and documented by fluorescent microscopy. For the IsoFlux system, best performance was obtained with IsoFlux CTC enrichment kit, but these beads compromised the subsequent immunofluorescence staining. For the KingFisher system, best recoveries were obtained using Dynabeads Biotin Binder beads. These beads also allowed one to capture CTCs with different antibodies and the subsequent immunofluorescence staining. KingFisher instrument allowed a single and streamlined protocol for the enrichment and staining of CTCs that further prevented cell loss at the enrichment/staining interface. Both IsoFlux and KingFisher systems allowed the enrichment of cell line cells from the mimicked-DLA samples. However, in this particular experimental setting, the recovery rates obtained with the KingFisher system were globally higher, the system was more cost-effective, and it allowed higher throughput. Full article
(This article belongs to the Special Issue Liquid Biopsy: Latest Advances and Future Challenges)
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15 pages, 4528 KiB  
Article
Detection of Circulating Tumor Cells in the Diagnostic Leukapheresis Product of Non-Small-Cell Lung Cancer Patients Comparing CellSearch® and ISET
by Menno Tamminga, Kiki C. Andree, T. Jeroen N. Hiltermann, Maximilien Jayat, Ed Schuuring, Hilda van den Bos, Diana C. J. Spierings, Peter M. Lansdorp, Wim Timens, Leon W. M. M. Terstappen and Harry J. M. Groen
Cancers 2020, 12(4), 896; https://doi.org/10.3390/cancers12040896 - 07 Apr 2020
Cited by 32 | Viewed by 4412
Abstract
Circulating tumor cells (CTCs) detected by CellSearch are prognostic in non-small-cell lung cancer (NSCLC), but rarely found. CTCs can be extracted from the blood together with mononuclear cell populations by diagnostic leukapheresis (DLA), therefore concentrating them. However, CellSearch can only process limited DLA [...] Read more.
Circulating tumor cells (CTCs) detected by CellSearch are prognostic in non-small-cell lung cancer (NSCLC), but rarely found. CTCs can be extracted from the blood together with mononuclear cell populations by diagnostic leukapheresis (DLA), therefore concentrating them. However, CellSearch can only process limited DLA volumes (≈2 mL). Therefore, we established a protocol to enumerate CTCs in DLA products with Isolation by SizE of Tumor cells (ISET), and compared CTC counts between CellSearch® and ISET. DLA was performed in NSCLC patients who started a new therapy. With an adapted protocol, ISET could process 10 mL of DLA. CellSearch detected CTCs in a volume equaling 2 × 108 leukocytes (mean 2 mL). CTC counts per mL were compared. Furthermore, the live cell protocol of ISET was tested in eight patients. ISET successfully processed all DLA products—16 with the fixed cell protocol and 8 with the live cell protocol. In total, 10–20 mL of DLA was processed. ISET detected CTCs in 88% (14/16), compared to 69% (11/16, p < 0.05) with CellSearch. ISET also detected higher number of CTCs (ISET median CTC/mL = 4, interquartile range [IQR] = 2–6, CellSearch median CTC/mL = 0.9, IQR = 0–1.8, p < 0.01). Cells positive for the epithelial cell adhesion molecule (EpCAM+) per mL were detected in similar counts by both methods. Eight patients were processed with the live cell protocol. All had EpCAM+, CD45−, CD235- cells isolated by fluorescence-activated cell sorting (FACS). Overall, ISET processed larger volumes and detected higher CTC counts compared to CellSearch. EpCAM+ CTCs were detected in comparable rates. Full article
(This article belongs to the Special Issue Liquid Biopsy: Latest Advances and Future Challenges)
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Review

Jump to: Research

20 pages, 633 KiB  
Review
Circulating Tumor Cells from Enumeration to Analysis: Current Challenges and Future Opportunities
by Yu-Ping Yang, Teresa M. Giret and Richard J. Cote
Cancers 2021, 13(11), 2723; https://doi.org/10.3390/cancers13112723 - 31 May 2021
Cited by 26 | Viewed by 5206
Abstract
Circulating tumor cells (CTCs) have been recognized as a major contributor to distant metastasis. Their unique role as metastatic seeds renders them a potential marker in the circulation for early cancer diagnosis and prognosis as well as monitoring of therapeutic response. In the [...] Read more.
Circulating tumor cells (CTCs) have been recognized as a major contributor to distant metastasis. Their unique role as metastatic seeds renders them a potential marker in the circulation for early cancer diagnosis and prognosis as well as monitoring of therapeutic response. In the past decade, researchers mainly focused on the development of isolation techniques for improving the recovery rate and purity of CTCs. These developed techniques have significantly increased the detection sensitivity and enumeration accuracy of CTCs. Currently, significant efforts have been made toward comprehensive molecular characterization, ex vivo expansion of CTCs, and understanding the interactions between CTCs and their associated cells (e.g., immune cells and stromal cells) in the circulation. In this review, we briefly summarize existing CTC isolation technologies and specifically focus on advances in downstream analysis of CTCs and their potential applications in precision medicine. We also discuss the current challenges and future opportunities in their clinical utilization. Full article
(This article belongs to the Special Issue Liquid Biopsy: Latest Advances and Future Challenges)
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24 pages, 1115 KiB  
Review
MicroRNAs from Liquid Biopsy Derived Extracellular Vesicles: Recent Advances in Detection and Characterization Methods
by Rares Drula, Leonie Florence Ott, Ioana Berindan-Neagoe, Klaus Pantel and George A. Calin
Cancers 2020, 12(8), 2009; https://doi.org/10.3390/cancers12082009 - 22 Jul 2020
Cited by 41 | Viewed by 5179
Abstract
Liquid biopsies have become a convenient tool in cancer diagnostics, real-time disease monitoring, and evaluation of residual disease. Yet, the information still encrypted in the variety of tumor-derived molecules identified in biofluids has proven difficult to decipher due to the technological limitations imposed [...] Read more.
Liquid biopsies have become a convenient tool in cancer diagnostics, real-time disease monitoring, and evaluation of residual disease. Yet, the information still encrypted in the variety of tumor-derived molecules identified in biofluids has proven difficult to decipher due to the technological limitations imposed by their biological nature. Such is the case of extracellular vesicle (EV) encapsulated ncRNAs, which have gained traction in recent years as biomarkers. Due to their resilience towards degrading factors they may act as suitable disease indicators. This review addresses the less described issues in this context. We present an overview of less investigated biofluids that can be used for EV isolation in addition to different isolation approaches to overcome the technical challenges these specimens harbor. Furthermore, we summarize the latest technological advances providing improvement to ncRNA detection and analysis. Thereby, this review summarizes the current state-of-the-art methodologies regarding EV and EV derived miRNA analysis and how they compare to current approaches. Full article
(This article belongs to the Special Issue Liquid Biopsy: Latest Advances and Future Challenges)
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16 pages, 473 KiB  
Review
The Role of Liquid Biopsies in Detecting Molecular Tumor Biomarkers in Brain Cancer Patients
by Heena Sareen, Celine Garrett, David Lynch, Branka Powter, Daniel Brungs, Adam Cooper, Joseph Po, Eng-Siew Koh, Joey Yusof Vessey, Simon McKechnie, Renata Bazina, Mark Sheridan, James van Gelder, Balsam Darwish, Mathias Jaeger, Tara L. Roberts, Paul De Souza and Therese M. Becker
Cancers 2020, 12(7), 1831; https://doi.org/10.3390/cancers12071831 - 08 Jul 2020
Cited by 31 | Viewed by 5144
Abstract
Glioblastoma multiforme (GBM) is one of the most lethal primary central nervous system cancers with a median overall survival of only 12–15 months. The best documented treatment is surgical tumor debulking followed by chemoradiation and adjuvant chemotherapy with temozolomide, but treatment resistance and [...] Read more.
Glioblastoma multiforme (GBM) is one of the most lethal primary central nervous system cancers with a median overall survival of only 12–15 months. The best documented treatment is surgical tumor debulking followed by chemoradiation and adjuvant chemotherapy with temozolomide, but treatment resistance and therefore tumor recurrence, is the usual outcome. Although advances in molecular subtyping suggests GBM can be classified into four subtypes, one concern about using the original histology for subsequent treatment decisions is that it only provides a static snapshot of heterogeneous tumors that may undergo longitudinal changes over time, especially under selective pressure of ongoing therapy. Liquid biopsies obtained from bodily fluids like blood and cerebro-spinal fluid (CSF) are less invasive, and more easily repeated than surgery. However, their deployment for patients with brain cancer is only emerging, and possibly suppressed clinically due to the ongoing belief that the blood brain barrier prevents the egress of circulating tumor cells, exosomes, and circulating tumor nucleic acids into the bloodstream. Although brain cancer liquid biopsy analyses appear indeed challenging, advances have been made and here we evaluate the current literature on the use of liquid biopsies for detection of clinically relevant biomarkers in GBM to aid diagnosis and prognostication. Full article
(This article belongs to the Special Issue Liquid Biopsy: Latest Advances and Future Challenges)
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25 pages, 875 KiB  
Review
Clinical Relevance of Liquid Biopsy in Melanoma and Merkel Cell Carcinoma
by Magali Boyer, Laure Cayrefourcq, Olivier Dereure, Laurent Meunier, Ondine Becquart and Catherine Alix-Panabières
Cancers 2020, 12(4), 960; https://doi.org/10.3390/cancers12040960 - 13 Apr 2020
Cited by 26 | Viewed by 6226
Abstract
Melanoma and Merkel cell carcinoma are two aggressive skin malignancies with high disease-related mortality and increasing incidence rates. Currently, invasive tumor tissue biopsy is the gold standard for their diagnosis, and no reliable easily accessible biomarker is available to monitor patients with melanoma [...] Read more.
Melanoma and Merkel cell carcinoma are two aggressive skin malignancies with high disease-related mortality and increasing incidence rates. Currently, invasive tumor tissue biopsy is the gold standard for their diagnosis, and no reliable easily accessible biomarker is available to monitor patients with melanoma or Merkel cell carcinoma during the disease course. In these last years, liquid biopsy has emerged as a candidate approach to overcome this limit and to identify biomarkers for early cancer diagnosis, prognosis, therapeutic response prediction, and patient follow-up. Liquid biopsy is a blood-based non-invasive procedure that allows the sequential analysis of circulating tumor cells, circulating cell-free and tumor DNA, and extracellular vesicles. These innovative biosources show similar features as the primary tumor from where they originated and represent an alternative to invasive solid tumor biopsy. In this review, the biology and technical challenges linked to the detection and analysis of the different circulating candidate biomarkers for melanoma and Merkel cell carcinoma are discussed as well as their clinical relevance. Full article
(This article belongs to the Special Issue Liquid Biopsy: Latest Advances and Future Challenges)
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20 pages, 1155 KiB  
Review
Gauging the Impact of Cancer Treatment Modalities on Circulating Tumor Cells (CTCs)
by Trevor J. Mathias, Katarina T. Chang, Stuart S. Martin and Michele I. Vitolo
Cancers 2020, 12(3), 743; https://doi.org/10.3390/cancers12030743 - 21 Mar 2020
Cited by 10 | Viewed by 3819
Abstract
The metastatic cascade consists of multiple complex steps, but the belief that it is a linear process is diminishing. In order to metastasize, cells must enter the blood vessels or body cavities (depending on the cancer type) via active or passive mechanisms. Once [...] Read more.
The metastatic cascade consists of multiple complex steps, but the belief that it is a linear process is diminishing. In order to metastasize, cells must enter the blood vessels or body cavities (depending on the cancer type) via active or passive mechanisms. Once in the bloodstream and/or lymphatics, these cancer cells are now termed circulating tumor cells (CTCs). CTC numbers as well as CTC clusters have been used as a prognostic marker with higher numbers of CTCs and/or CTC clusters correlating with an unfavorable prognosis. However, we have very limited knowledge about CTC biology, including which of these cells are ultimately responsible for overt metastatic growth, but due to the fact that higher numbers of CTCs correlate with a worse prognosis; it would seem appropriate to either limit CTCs and/or their dissemination. Here, we will discuss the different cancer treatments which may inadvertently promote the mobilization of CTCs and potential CTC therapies to decrease metastasis. Full article
(This article belongs to the Special Issue Liquid Biopsy: Latest Advances and Future Challenges)
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