Ex Vivo Expansion and Drug Sensitivity Profiling of Circulating Tumor Cells from Patients with Small Cell Lung Cancer

Abstract

Small cell lung cancer (SCLC) represents one of the most aggressive malignancies among cancer types. Not only tumor sample availability is limited, but also the ability for tumor cells to rapidly acquire drug resistance are the rate-limiting bottlenecks for overall survival in current clinical settings. A liquid biopsy capable of capturing and enriching circulating tumor cells (CTCs), together with the possibility of drug screening, is a promising solution. Here, we illustrate the development of a highly efficient ex vivo CTC expansion system based on binary colloidal crystals substrate. Clinical samples were enrolled from 22 patients with SCLC in the study. The CTCs were enriched and expanded from the collected peripheral blood samples. Expanded cells were analyzed for protein expression and observed for drug sensitivity with the use of immunofluorescence and ATP titer evaluation, respectively. Successful CTC spheroid proliferation was established after 4 weeks within 82% of all the collected peripheral blood samples from enrolled patients. Upon immunofluorescence analysis, the enriched cells showed positive markers for EpCAM, TTF-1, synaptophysin and negative for CD45. Additionally, the expanded CTCs demonstrated marked heterogeneity in the expression of E-cadherin and N-cadherin. In a preliminary case series, the drug sensitivity of patient-derived CTC to cisplatin and etoposide was studied to see the correlation with the corresponding therapeutic outcome. In conclusion, our study demonstrates that it is possible to efficiently expand CTCs from SCLC within a clinically relevant time frame; the biomarker information generated from enriched CTCs can assist the selection of effective drugs and improve disease outcome.

Patient-Derived Tumor Chemosensitization of GKB202, an Antrodia Cinnamomea Mycelium-Derived Bioactive Compound

“…Recently, our group developed a CTC organoid culture workflow that efficiently expand CTC from patients with extensive stage small cell lung cancer [ 21 ]. Moreover, the drug sensitivity profile from these CTC-derived organoids correlates well to the clinical treatment response.…”

Clinical Relevance of Mesenchymal- and Stem-Associated Phenotypes in Circulating Tumor Cells Isolated from Lung Cancer Patients

“…Ex vivo expansion of CTCs from patients with SCLC has also been recently [ 74 ] reported. Initial findings have reported the existence of EMT and of other currently investigated markers such as PD-L1, but this will have to be further examined.…”

Correlation Between Drug Sensitivity Profiles of Ex Vivo Expanded Circulating Tumor Cells and Clinical Treatment Response in Pancreatic Ductal Adenocarcinoma Patients

“…Liquid biopsies consisting of a 20-mL sample of peripheral venous blood were collected from each patient, using K2EDTA Vacutainers® (BD Bioscience, Franklin Lakes NJ, USA). A total of 41 liquid biopsies were collected, from which the peripheral blood mononuclear cell (PBMC) fraction containing CTCs was extracted using Ficoll-Paque centrifugation as previously described [17,20]. Cells were then seeded onto a substrate of binary colloidal crystal (BCC) containing silica and polymethyl methacrylate (PMMA), and cultured in the eSelect system for 3 weeks, with replacement of culture medium every 4 days.…”

“…Cells were then seeded onto a substrate of binary colloidal crystal (BCC) containing silica and polymethyl methacrylate (PMMA), and cultured in the eSelect system for 3 weeks, with replacement of culture medium every 4 days. Organoid expansion was monitored by visual microcopy, and the presence of CTCs was con rmed by EpCAM and CD45 immuno uorescence staining in a procedure described previously [17]. Patients for whom CTC-derived organoid expansion was unsuccessful were denoted as "failed expansion.…”

“…These CTC-derived organoid cultures were assessed for sensitivity to a panel of nine anti-cancer drugs, and the results were correlated with clinical response in patients over a 3-month followup period. A previous study using the eSelect system showed good correlation between the drug sensitivity pro les of CTC-derived organoid cultures and clinical response to treatment in small cell lung cancer patients [17], and demonstrated the potential to reduce costs and time involved with culturing and testing. Here we showed that drug sensitivity results derived from CTC-derived organoids, calculated as a cytotoxicity score (CTS), correlated signi cantly with clinical outcomes in prospective utility analysis.…”