Circulating Tumor Cells in Cancer Diagnostics and Prognostics by Single-Molecule and Single-Cell Characterization

Abstract

Circulating tumor cells (CTCs) represent an interesting source of biomarkers for diagnosis, prognosis, and the prediction of cancer recurrence, yet while they are extensively studied in oncobiology research, their diagnostic utility has not yet been demonstrated and validated. Their scarcity in human biological fluids impedes the identification of dangerous CTC subpopulations that may promote metastatic dissemination. In this Perspective, we discuss promising techniques that could be used for the identification of these metastatic cells. We first describe methods for isolating patient-derived CTCs and then the use of 3D biomimetic matrixes in their amplification and analysis, followed by methods for further CTC analyses at the single-cell and single-molecule levels. Finally, we discuss how the elucidation of mechanical and morphological properties using techniques such as atomic force microscopy and molecular biomarker identification using nanopore-based detection could be combined in the future to provide patients and their healthcare providers with a more accurate diagnosis.

Integrated Microfluidics for Single‐Cell Separation and On‐Chip Analysis: Novel Applications and Recent Advances

“…The analysis of CTCs presents distinct challenges due to their scarcity in the bloodstream (estimated 1-10 CTCs mL À1 of blood) and the lack of specific markers indicative of metastasis-promoting cells. [114,183,184] Furthermore, CTCs may exist as clusters rather than single cells, complicating their handling in microfluidic devices. [185,186] Lastly, the expectation for Codetection of multiple secreted factors in relation to cell proximity and migration behavior [202] Human oral squamous carcinoma cells and human primary oral squamous carcinoma cells were suspended in cell culture medium; human primary CAF cells were isolated from patient samples and suspended in cell culture medium, human primary monocytes were isolated from PBMCs and differentiated into macrophages -Microchambers Biological analysis (morphology, migration)…”

Detection and Molecular Characterization of Circulating Tumour Cells: Challenges for the Clinical Setting

“…Thus, a high variety of novel ‘label-free’ CTC enrichment technologies based on CTC size and deformability have been developed [ 4 ]. These technologies can be categorized as: (i) systems based on a combination of microfluidics and size-based enrichment, such as the Parsortix system, (Angle, UK), that was FDA-cleared in May 2022 for CTC enrichment in metastatic breast cancer (MBC), (ii) filtration devices based on different CTC size, such as the ScreenCell device (Screencell, Paris, France) and the ISET system (RareCells, France), (iii) microchips with integrated nanomaterials, such as the CTC chip (NP-HBCTC-Chip) and the graphene oxide (GO) chip, (iv) systems for single CTC isolation, such as the DepArray (Menarini, Silicon Biosystems, Italy), the VyCAP (VyCAP BV, The Netherlands), and the CellCollector (ALS, Sartorius), (v) epitope-agnostic systems that detect both EpCAM-pos and EpCAM-neg CTCs, such as the EPISPOT, and the Epic platform (Epic Sciences), (vi) density-based isolation followed by antibody cocktail for imaging (RareCyte system, Rarecyte, US), and (vii) molecular tests, such as the Adnatest (Qiagen, Germany), and many others [ 1 , 2 , 3 , 5 , 6 , 7 , 8 , 9 , 10 , 11 ].…”

3D Printing for Cancer Diagnosis: What Unique Advantages Are Gained?

“…Among these advancements, the emergence of 3D printing technology has revolutionized the field of cancer diagnostics, offering immense potential in detecting and monitoring the disease. 4−7 Cancer is characterized by the uncontrolled growth and spread of abnormal cells within the body 8,9 Early detection plays a crucial role in improving patient outcomes and survival rates. Traditional diagnostic methods, such as imaging techniques (e.g., X-ray, 10 computed tomography, 11,12 magnetic resonance imaging 13,14 ) and biopsy, have been instrumental in cancer diagnosis.…”

“…Cancer is characterized by the uncontrolled growth and spread of abnormal cells within the body , Early detection plays a crucial role in improving patient outcomes and survival rates. Traditional diagnostic methods, such as imaging techniques (e.g., X-ray, computed tomography, , magnetic resonance imaging , ) and biopsy, have been instrumental in cancer diagnosis.…”