Covalent Chemistry‐Mediated Multimarker Purification of Circulating Tumor Cells Enables Noninvasive Detection of Molecular Signatures of Hepatocellular Carcinoma

Abstract

Transcriptomic profiling of tumor tissues introduces a large database, which has led to improvements in the ability of cancer diagnosis, treatment, and prevention. However, performing tumor transcriptomic profiling in the clinical setting is very challenging since the procurement of tumor tissues is inherently limited by invasive sampling procedures. Here, the feasibility of purifying hepatocellular carcinoma (HCC) circulating tumor cells (CTCs) from clinical patient samples is demonstrated with improved molecular integrity using Click Chips in conjunction with a multimarker antibody cocktail. The purified CTCs are then subjected to messenger RNA profiling by NanoString nCounter platform, targeting 64 HCC‐specific genes, which are generated from an integrated data analysis framework with eight tissue‐based prognostic gene signatures from seven publicly available HCC transcriptomic studies. After bioinformatics analysis and comparison, the HCC CTC‐derived gene signatures show high concordance with HCC tissue‐derived gene signatures from the Cancer Genome Atlas database, suggesting that HCC CTCs purified by Click Chips can enable the translation of HCC tissue molecular profiling into a noninvasive setting.

Circulating tumor cells: A step toward precision medicine in hepatocellular carcinoma

“…Sun et al . displayed that the CTCs purified by the latest NanoVelcro assay, that is, Click Chip, possessed highly concordant expression of 64 HCC tissue‐derived genes from The Cancer Genome Atlas (TCGA) 14 . Lee et al .…”

“…62 Sun et al displayed that the CTCs purified by the latest NanoVelcro assay, that is, Click Chip, possessed highly concordant expression of 64 HCC tissue-derived genes from The Cancer Genome Atlas (TCGA). 14 Lee et al established a 10-gene HCC CTC Risk Score panel (DDR1, EHHADH, AR, LUM, HSD17B6, PMEPA1, TSKU, NECAB2, LAD1, and SLC27A5) quantified by the NanoString nCounter platform 65 and the risk score was significantly associated with overall survival. 15 Besides serving as a biomarker assay, many researchers also utilized CTC platforms to explore cancer biology.…”

“…The androgen receptor splice variant 7 (AR‐V7) RNA expression in prostate cancer CTCs was validated to have values in predicting hormonal therapy response in a multicenter, prospective‐blinded clinical trial, and the results were consistent between two CTC platforms of different enrichment mechanisms 11 . Some researchers might argue that a single‐gene expression is biased and insufficient for prognostication, so several studies have established CTC‐specific gene signatures to provide comprehensive biological insights 12–15 . In this review article, we will briefly introduce the existing CTC technologies and focus on discussing clinically relevant studies with an adequate sample size in HCC.…”

“…11 Some researchers might argue that a single-gene expression is biased and insufficient for prognostication, so several studies have established CTC-specific gene signatures to provide comprehensive biological insights. [12][13][14][15] In this review article, we will briefly introduce the existing CTC technologies and focus on discussing clinically relevant studies with an adequate sample size in HCC.…”

Hierarchical integration of DNA nanostructures and NanoGold onto a microchip facilitates covalent chemistry-mediated purification of circulating tumor cells in head and neck squamous cell carcinoma

“…By taking advantage of this unique feature, we and others have pioneered the use of nanostructured substrates for enrichment, isolation, and characterization of rare cells, including, but not limited to, circulating tumor cells (CTCs) [ 6 ] and circulating fetal nucleated cells (CFNCs) [ 10 – 12 ]. Particularly, NanoVelcro Chips [ 13 – 15 ] developed by our research team have been applied to various clinical needs such as CTC enumeration [ 16 ], single-cell analysis [ 17 ], and CTC purification [ 18 , 19 ] with well-preserved RNA and cell viability. Though it has been successfully demonstrated that the previously developed nanostructured substrates enrich rare cells by relying on the nanostructure-mediated cell-affinity capture, all of these methods are based on the integration of a single layer of nanostructure onto a supporting substrate.…”