The incorporation of microfluidics into circulating tumor cell isolation for clinical applications

Kozminsky, Molly; Wang, Yan; Nagrath, Sunitha

doi:10.1016/j.coche.2016.01.005

Cited by 13 publications

(13 citation statements)

References 45 publications

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“…The clinical significance of CTCs has been shown in multiple cancers, including prostate cancer, in which the prognostic values of CTC counts above and below the cut-off of 5 CTCs/7.5 mL whole blood have been associated with overall survival (15). Beyond enumeration, novel CTC isolation technologies have also enabled further characterization, including protein expression by immunofluorescence (16,17). …”

Section: Introductionmentioning

confidence: 99%

HER2 and EGFR Overexpression Support Metastatic Progression of Prostate Cancer to Bone

et al. 2017

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Activation of the epidermal growth factor receptors EGFR (ErbB1) and HER2 (ErbB2) drive the progression of multiple cancer types through complex mechanisms that are still not fully understood. In this study, we report that HER2 expression is elevated in bone metastases of prostate cancer independently of gene amplification. An examination of HER2 and NF-κB receptor (RANK) coexpression revealed increased levels of both proteins in aggressive prostate tumors and metastatic deposits. Inhibiting HER2 expression in bone tumor xenografts reduced proliferation and RANK expression while maintaining EGFR expression. In examining the role of EGFR in tumor-initiating cells (TIC), we found that EGFR expression was required for primary and secondary sphere formation of prostate cancer cells. EGFR expression was also observed in circulating tumor cells (CTC) during prostate cancer metastasis. Dual inhibition of HER2 and EGFR resulted in significant inhibition of tumor xenograft growth, further supporting the significance of these receptors in prostate cancer progression. Overall, our results indicate that EGFR promotes survival of prostate TIC and CTC that metastasize to bone, whereas HER2 supports the growth of prostate cancer cells once they are established at metastatic sites.

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Section: Introductionmentioning

confidence: 99%

HER2 and EGFR Overexpression Support Metastatic Progression of Prostate Cancer to Bone

et al. 2017

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“…This problem has been addressed with a host of isolation technologies . Notably, the first FDA‐approved CTC isolation technology, CellSearch, has been used to establish survival differences based on CTC enumeration .…”

Section: Introductionmentioning

confidence: 99%

Detection of CTC Clusters and a Dedifferentiated RNA‐Expression Survival Signature in Prostate Cancer

et al. 2018

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Rates of progression and treatment response in advanced prostate cancer are highly variable, necessitating non‐invasive methods to assess the molecular characteristics of these tumors in real time. The unique potential of circulating tumor cells (CTCs) to serve as a clinically useful liquid biomarker is due to their ability to inform via both enumeration and RNA expression. A microfluidic graphene oxide‐based device (GO Chip) is used to isolate CTCs and CTC clusters from the whole blood of 41 men with metastatic castration‐resistant prostate cancer. Additionally, the expression of 96 genes of interest is determined by RT‐qPCR. Multivariate analyses are conducted to determine the genes most closely associated with overall survival, PSA progression, and radioclinical progression. A preliminary signature, comprising high expression of stemness genes and low expression of epithelial and mesenchymal genes, potentially implicates an undifferentiated CTC phenotype as a marker of poor prognosis in this setting.

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“…El-Heliebi et al compared three different CTC isolation systems for gene expression and DNA mutation analysis in CTCs of prostate cancer patients and showed also differences 32 . The incorporation of microfluidics into CTC isolation is now emerging for clinical applications 33 . Many microfluidic technologies have reported high sensitivity and specificity for capturing CTCs, however, the question still remains as to the superiority in comparison to immunoaffinity based approaches, specifically to identify different CTC populations 34 .…”

Section: Discussionmentioning

confidence: 99%

Direct comparison of size-dependent versus EpCAM-dependent CTC enrichment at the gene expression and DNA methylation level in head and neck squamous cell carcinoma

Zavridou

Mastoraki

Strati

et al. 2020

Sci Rep

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ANGLE) and an epcAM-dependent positive immune-magnetic isolation procedure were applied in parallel, using 10 mL PB from 50 HNSCC patients and 18 healthy donors. Total RNA was isolated from enriched CTCs and Rt-qpcR was used to study the expression levels of CK-19, PD-L1, EGFR, TWIST1, CDH2 and B2M (reference gene). Real time methylation specific PCR (MSP) was used to study the methylation status of RASSF1A and MLL3 genes. In identical blood draws, the label-free size-dependent CTC-isolation system was superior in terms of sensitivity when compared to the epcAM-dependent ctc enrichment, since a significantly higher percentage of identical PB samples was found positive at the gene expression and DNA methylation level, while the specificity was not affected. Our results indicate that future studies focused on the evaluation of clinical utility of ctc molecular characterization in HnScc should be based on size-dependent enrichment approaches.Liquid biopsy provides a valuable source of biomarkers on prognosis and response to treatment of cancer patients 1 and has recently shown a significant potential even for early cancer diagnosis and screening 2 . Isolation of circulating tumor cells (CTCs) from peripheral blood (PB) and their further downstream molecular characterization at the DNA, RNA and protein level is very important for reliable liquid biopsy analysis 3 . However, the identification and molecular characterization of CTCs is very challenging since these cells are extremely rare, and the amount of available sample for analysis in most cases is very limited 1,3 .A variety of molecular assays have been developed for CTCs detection and molecular characterization. Molecular assays are based on total RNA isolation from CTCs and subsequent mRNA quantification of specific genes, and gDNA isolation for mutation analysis and DNA methylation studies 4 . CTC molecular characterization at the gene expression level has the potential to elucidate the critical signaling pathways involved in metastasis biology and even improve patient management. We have shown many years ago that the detection of CK-19 expression in CTCs has prognostic significance in both early and metastatic breast cancer 5-7 . Beyond gene expression, DNA methylation analysis in CTCs has a high potential to provide novel epigenetic biomarkers for diagnosis, prognosis, risk assessment, and disease monitoring in many types of cancer 4 . Based on this, we selected www.nature.com/scientificreports www.nature.com/scientificreports/ gDNA isolation from CTCs. gDNA was extracted from CTCs using the TRIZOL-LS reagent (ThermoFisher Scientific, USA) as previously described. Isolated gDNA 44 was dissolved in 30 μL of 8 mmol/L NaOH. Sodium Bisulfite (SB) treatment. gDNA samples were treated with SB, to convert all non-methylated cytosines to uracil, while methylated cytosines were not converted, using the EZ DNA Methylation Gold Kit (ZYMO Research, USA). SB-treated DNA was stored at −70 °C until use. In each SB reaction, dH 2 O and 100% methylated DNA were included as ...

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The incorporation of microfluidics into circulating tumor cell isolation for clinical applications

Cited by 13 publications

References 45 publications

HER2 and EGFR Overexpression Support Metastatic Progression of Prostate Cancer to Bone

HER2 and EGFR Overexpression Support Metastatic Progression of Prostate Cancer to Bone

Detection of CTC Clusters and a Dedifferentiated RNA‐Expression Survival Signature in Prostate Cancer

Direct comparison of size-dependent versus EpCAM-dependent CTC enrichment at the gene expression and DNA methylation level in head and neck squamous cell carcinoma

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