Exploration of Circulating Tumour Cell (CTC) Biology: A Paradigm Shift in Liquid Biopsy

Chauhan, Anshika; Kaur, Rajandeep; Ghoshal, Sushmita

doi:10.1007/s12291-020-00923-4

Cited by 13 publications

(7 citation statements)

References 93 publications

(97 reference statements)

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“…On the contrary, analysis of CTCs provides a minimally invasive and easily repeatable approach for real-time monitoring of the disease process, which is essential for treatment success. As tumour evolves spatially and temporally ( 16 ), it is more important to analyse the molecular features of CTCs rather than their detection or enumeration ( 17 ). CTCs might represent the entire spectrum of clones in a solid tumour at a specific time, suggesting heterogeneity within the CTC population.…”

Section: Discussionmentioning

confidence: 99%

A FACS-based novel isolation technique identifies heterogeneous CTCs in oral squamous cell carcinoma

Chauhan,

Pal,

Sachdeva

et al. 2024

Front. Oncol.

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PurposeIsolating circulating tumour cells (CTCs) from the blood is challenging due to their low abundance and heterogeneity. Limitations of conventional CTC detection methods highlight the need for improved strategies to detect and isolate CTCs. Currently, the Food and Drug Administration (FDA)-approved CellSearch™ and other RUO techniques are not available in India. Therefore, we wanted to develop a flexible CTC detection/isolation technique that addresses the limitation(s) of currently available techniques and is suitable for various downstream applications.MethodsWe developed a novel, efficient, user-friendly CTC isolation strategy combining density gradient centrifugation and immuno-magnetic hematogenous cell depletion with fluorescence-activated cell sorting (FACS)-based positive selection using multiple CTC-specific cell-surface markers. For FACS, a stringent gating strategy was optimised to exclude debris and doublets by side scatter/forward scatter (SSC/FSC) discriminator, remove dead cells by 4′,6-diamidino-2-phenylindole (DAPI) staining, and eliminate non-specific fluorescence using a “dump” channel. APC-labelled anti-CD45mAB was used to gate remaining hematogenous cells, while multiple epithelial markers (EpCAM, EGFR, and Pan-Cytokeratin) and an epithelial–mesenchymal transition (EMT) marker (Vimentin) labelled with fluorescein isothiocyanate (FITC) were used to sort cancer cells. The technique was initially developed by spiking Cal 27 cancer cells into the blood of healthy donors and then validated in 95 biopsy-proven oral squamous cell carcinoma (OSCC) patients. CTCs isolated from patients were reconfirmed by Giemsa staining, immuno-staining, and whole transcriptome amplification (WTA), followed by qRT-PCR. In vitro culture and RNA sequencing (RNA-Seq) were also performed to confirm their suitability for various downstream applications.ResultsThe mean detection efficiency for the Cal 27 tongue cancer cells spiked in the whole blood of healthy donors was 32.82% ± 12.71%. While ~75% of our patients (71/95) had detectable CTCs, the CTC positivity was independent of the TNM staging. The isolated potential cancer cells from OSCC patients were heterogeneous in size. They expressed different CTC-specific markers in various combinations as identified by qRT-PCR after WTA in different patients. Isolated CTCs were also found to be suitable for downstream applications like short-term CTC culture and RNA-Seq.ConclusionWe developed a sensitive, specific, flexible, and affordable CTC detection/isolation technique, which is scalable to larger patient cohorts, provides a snapshot of CTC heterogeneity, isolates live CTCs ready for downstream molecular analysis, and, most importantly, is suitable for developing countries.

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Section: Discussionmentioning

confidence: 99%

A FACS-based novel isolation technique identifies heterogeneous CTCs in oral squamous cell carcinoma

Chauhan,

Pal,

Sachdeva

et al. 2024

Front. Oncol.

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“…Circulating tumor cells are tumorous cells that exuviate from primary or metastatic tumors and extravasate to enter and stay in the blood and lymph circulation. These rare cells travel throughout circulation to eventually colonize a distant organ and form secondary tumors (metastasis) ( Chauhan et al, 2021 ) ( Figure 2 ). They exhibit considerable heterogeneity in terms of size, shape, and surface characteristics.…”

Section: Circulating Tumor Cellsmentioning

confidence: 99%

Prospects of liquid biopsy in the prognosis and clinical management of gastrointestinal cancers

Mondal,

Shinde,

Sinha

et al. 2024

Front. Mol. Biosci.

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Gastrointestinal (GI) cancers account for one-fourth of the global cancer incidence and are incriminated to cause one-third of cancer-related deaths. GI cancer includes esophageal, gastric, liver, pancreatic, and colorectal cancers, mostly diagnosed at advanced stages due to a lack of accurate markers for early stages. The invasiveness of diagnostic methods like colonoscopy for solid biopsy reduces patient compliance as it cannot be frequently used to screen patients. Therefore, minimally invasive approaches like liquid biopsy may be explored for screening and early identification of gastrointestinal cancers. Liquid biopsy involves the qualitative and quantitative determination of certain cancer-specific biomarkers in body fluids such as blood, serum, saliva, and urine to predict disease progression, therapeutic tolerance, toxicities, and recurrence by evaluating minimal residual disease and its correlation with other clinical features. In this review, we deliberate upon various tumor-specific cellular and molecular entities such as circulating tumor cells (CTCs), tumor-educated platelets (TEPs), circulating tumor DNA (ctDNA), cell-free DNA (cfDNA), exosomes, and exosome-derived biomolecules and cite recent advances pertaining to their use in predicting disease progression, therapy response, or risk of relapse. We also discuss the technical challenges associated with translating liquid biopsy into clinical settings for various clinical applications in gastrointestinal cancers.

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“…Moreover, the downregulation of epithelial markers like EpCAM might result from EMT. 147 In this phase, applying EpCAM-based selection methods does not allow for an accurate estimation of the number of CTCs. Therefore, simultaneously detecting epithelial and mesenchymal markers and adopting marker-free detection methods can improve CTC characterization efficiency.…”

Section: Immunophenotypic Characterization Of Ctcs and Ctc Clustersmentioning

confidence: 99%

Advances in novel strategies for isolation, characterization, and analysis of CTCs and ctDNA

Yaghoubi Naei,

Bordhan,

Mirakhorli

et al. 2023

Ther Adv Med Oncol

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Over the past decade, the detection and analysis of liquid biopsy biomarkers such as circulating tumor cells (CTCs) and circulating tumor DNA (ctDNA) have advanced significantly. They have received recognition for their clinical usefulness in detecting cancer at an early stage, monitoring disease, and evaluating treatment response. The emergence of liquid biopsy has been a helpful development, as it offers a minimally invasive, rapid, real-time monitoring, and possible alternative to traditional tissue biopsies. In resource-limited settings, the ideal platform for liquid biopsy should not only extract more CTCs or ctDNA from a minimal sample volume but also accurately represent the molecular heterogeneity of the patient’s disease. This review covers novel strategies and advancements in CTC and ctDNA-based liquid biopsy platforms, including microfluidic applications and comprehensive analysis of molecular complexity. We discuss these systems’ operational principles and performance efficiencies, as well as future opportunities and challenges for their implementation in clinical settings. In addition, we emphasize the importance of integrated platforms that incorporate machine learning and artificial intelligence in accurate liquid biopsy detection systems, which can greatly improve cancer management and enable precision diagnostics.

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Exploration of Circulating Tumour Cell (CTC) Biology: A Paradigm Shift in Liquid Biopsy

Cited by 13 publications

References 93 publications

A FACS-based novel isolation technique identifies heterogeneous CTCs in oral squamous cell carcinoma

A FACS-based novel isolation technique identifies heterogeneous CTCs in oral squamous cell carcinoma

Prospects of liquid biopsy in the prognosis and clinical management of gastrointestinal cancers

Advances in novel strategies for isolation, characterization, and analysis of CTCs and ctDNA

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