Novel Peptide-Based Magnetic Nanoparticle for Mesenchymal Circulating Tumor Cells Detection

Abstract

The monitoring of circulating tumor cells (CTCs) has recently served as a promising approach for assessing prognosis and evaluating cancer treatment. We have already developed a CTCs enrichment platform by EpCAM recognition peptide-functionalized magnetic nanoparticles (EP@MNPs). However, considering heterogeneous CTCs generated through epithelial-mesenchymal transition (EMT), mesenchymal CTCs would be missed with this method. Notably, N-cadherin, overexpressed on mesenchymal CTCs, can facilitate the migration of cancer cells. Hence, we screened a novel peptide targeting N-cadherin, NP, and developed a new CTCs isolation approach via NP@MNPs to complement EpCAM methods’ deficiencies. NP@MNPs had a high capture efficiency (about 85%) of mesenchymal CTCs from spiked human blood. Subsequently, CTCs were captured and sequenced at the single-cell level via NP@MNPs and EP@MNPs, RNA profiles of which showed that epithelial and mesenchymal subgroups could be distinguished. Here, a novel CTCs isolation platform laid the foundation for mesenchymal CTCs isolation and subsequent molecular analysis.

The Discovery of Novel Circulating Cancer‐Related Cells in Circulation Poses New Challenges to Microfluidic Devices for Enrichment and Detection

“…However, this system uses magnetic beads that only label EpCAM, which fails to capture the mesenchymal CTCs subset, resulting in a high rate of missed detection. [ 81 ] According to this reason, later studies began to use other markers (e.g., HER2, PSMA, PD‐L1), [ 82,83 ] or even adopt a multi‐marker strategy for the development of affinity‐based microfluidic devices to increase the capture efficiency ( Figure A). [ 16–18,87,88 ] Other studies began to focus on the binding efficiency between cell surface antigen and fixed antibody by geometrically enhanced mixing to increase the contact probability between cells and antibody‐functionalized surfaces, [ 89,90 ] or by developing nanostructures, [ 91 ] and magnetic particles.…”

Genomic Landscape of Liquid Biopsy for Hepatocellular Carcinoma Personalized Medicine

“…Different approaches for CTC isolation via targeting EMT markers have been proposed (66). In addition to CK19 and compared to EpCAM and vimentin, glypican-3 (GPC-3) has been proposed for CTCs separation, since a correlation between the positive count of CTCs using GPC-3 (≥5 CTC per 7.5 ml blood) and BCLC stage has been observed (67).…”

Engineering Peptide-Functionalized Biomimetic Nanointerfaces for Synergetic Capture of Circulating Tumor Cells in an EpCAM-Independent Manner

“…Aiming at recognizing CTCs, various affinity approaches have been developed based on the physical and biological properties of cancer cells. − A diversity of nanomaterials and devices capable of discriminating size or surface roughness between CTCs and blood cells has emerged for CTC capture. − To target CTCs at the molecular level, antibodies, aptamers, and peptides were employed as the capture element. − Epithelial markers, such as the epithelial cell adhesion molecule (EpCAM), are the most commonly used targets for designing recognition elements and CTC analysis. − Recently, strategies combining size selection and molecular recognition emerged with improved specificity and sensitivity for CTC screening. − Despite great advances, there are still hurdles limiting the efficiency and accuracy of CTC detection in real samples, including the heterogeneous nature of CTCs, the lack of specific and sensitive markers, and the loss of commonly used signatures during the epithelial–mesenchymal transition (EMT) process. ,, For tumor cells with high metastatic potential, epithelial markers (e.g., EpCAM) undergo downregulation, making these key subpopulations of CTCs undetectable by epithelial marker-based approaches. ,, Moreover, some tumor cells such as stem cell-like CTCs, cells from hepatocellular carcinoma (HCC), and soft tissue tumors express fewer epithelial markers and thus are insensitive to commonly used methods. For example, more than 80% of HCC patients are negative for EpCAM and hard to be detected by EpCAM-based approaches. − Besides, isolation of CTCs without affecting their viability and biological activity is a prerequisite for downstream investigation. , …”

“…18−20 Despite great advances, there are still hurdles limiting the efficiency and accuracy of CTC detection in real samples, including the heterogeneous nature of CTCs, the lack of specific and sensitive markers, and the loss of commonly used signatures during the epithelial− mesenchymal transition (EMT) process. 1,21,22 For tumor cells with high metastatic potential, epithelial markers (e.g., EpCAM) undergo downregulation, making these key subpopulations of CTCs undetectable by epithelial marker-based approaches. 1,23,24 Moreover, some tumor cells such as stem cell-like CTCs, cells from hepatocellular carcinoma (HCC), and soft tissue tumors express fewer epithelial markers and thus are insensitive to commonly used methods.…”