Multigene model for predicting metastatic prostate cancer using circulating tumor cells by microfluidic magnetophoresis

Abstract

We aimed to isolate circulating tumor cells (CTCs) using a microfluidic technique with a novel lateral magnetophoretic microseparator. Prostate cancer–specific gene expressions were evaluated using mRNA from the isolated CTCs. A CTC‐based multigene model was then developed for identifying advanced prostate cancer. Peripheral blood samples were obtained from five healthy donors and patients with localized prostate cancer (26 cases), metastatic hormone‐sensitive prostate cancer (mHSPC, 10 cases), and metastatic castration‐resistant prostate cancer (mCRPC, 28 cases). CTC recovery rate and purity (enriched CTCs/total cells) were evaluated according to cancer stage. The areas under the curves of the six gene expressions were used to evaluate whether multigene models could identify mHSPC or mCRPC. The number of CTCs and their purity increased at more advanced cancer stages. In mHSPC/mCRPC cases, the specimens had an average of 27.5 CTCs/mL blood, which was 4.2 × higher than the isolation rate for localized disease. The CTC purity increased from 2.1% for localized disease to 3.8% for mHSPC and 6.7% for mCRPC, with increased CTC expression of the genes encoding prostate‐specific antigen (PSA), prostate‐specific membrane antigen (PSMA), and cytokeratin 19 (KRT19). All disease stages exhibited expression of the genes encoding androgen receptor (AR) and epithelial cell adhesion molecule (EpCAM), although expression of the AR‐V7 variant was relatively rare. Relative to each gene alone, the multigene model had better accuracy for predicting advanced prostate cancer. Our lateral magnetophoretic microseparator can be used for identifying prostate cancer biomarkers. In addition, CTC‐based genetic signatures may guide the early diagnosis of advanced prostate cancer.

Perioperative Circulating Tumor Cells (CTCs), MCTCs, and CTC-White Blood Cells Detected by a Size-Based Platform Predict Prognosis in Renal Cell Carcinoma

“…As mentioned previously, CTCs in the bloodstream can be classified into three types, and their detection relied on a combination of membrane filtration and epithelial/mesenchymal biomarker-based identification. Recent reports indicated that the expression of EMT markers in CTCs is relevant process for invasion and metastasis in several cancers, such as breast, colorectal, nonsmall cell lung, gastric, and prostate cancers [ 26 – 30 ]. The CTC detection method in the present study allowed the isolation of pivotal EMT CTCs in renal cancer.…”

Circulating tumor cell gene expression and plasma AR gene copy number as biomarkers for castration-resistant prostate cancer patients treated with cabazitaxel

“…CTC positivity for the examined patient cohort was defined as the expression of at least 1 of the following 12 biomarkers: AR-V7, AKR1C3, AKT2, ALDH1, AR, EPCAM, PSMA, MDK, PIK3CA, PSCA, TUBB3, VIM. These biomarkers were selected based on literature data supporting their capacity to identify CTC as positive in prostate cancer or confirming their role in prostate cancer or in EMT [ 28 – 30 ].…”

Association of serum prostate-specific antigen (PSA) level and circulating tumor cell-based PSA mRNA in prostate cancer

“…Since the discovery that androgen receptor (AR)-targeting agents are closely related to the AR splice variant expressed on CTCs in advanced prostate cancer, CTC-based genomic analysis has been highlighted. 11 , 12 , 13 However, genomic analysis of CTCs remains challenging despite the development of novel techniques such as next-generation sequencing and droplet digital PCR (ddPCR) because of the presence of trace amounts of CTCs in blood. Recently, technology capable of separating CTCs more effectively has been developed and can be used for identifying biomarkers in PCa.…”

“…Recently, technology capable of separating CTCs more effectively has been developed and can be used for identifying biomarkers in PCa. 13 , 14 , 15 …”

“…Previous work has shown that many CTC based molecules can be used as a marker to predict the treatment response or prognosis of patients with PCa. 11 , 12 , 13 Among them, PSA mRNA is a very unique molecule. Since PSA mRNA is produced in epithelial cells of the prostate, it cannot be detected in the blood of men without PCa.…”