Feasibility study of in vitro drug sensitivity assay of advanced non-small cell lung adenocarcinomas

Background Despite improved screening techniques, diagnosis of lung cancer is often late and its prognosis is poor. In the present study, in vitro chemosensitivity of solid tumours and pleural effusions of lung adenocarcinomas were analysed and compared with clinical drug response.Methods Tumour cells were isolated from resected solid tumours or pleural effusions, and cryopreserved. Three-dimensional (3D) tissue aggregate cultures were set up when the oncoteam reached therapy decision for individual patients. The aggregates were then treated with the selected drug or drug combination and in vitro chemosensitivity was tested individually measuring ATP levels. The clinical response to therapy was assessed by standard clinical evaluation over an 18 months period.Results Based on the data, the in vitro chemosensitivity test results correlate well with clinical treatment response.Conclusions Such tests if implemented into the clinical decision making process might allow the selection of an even more individualised chemotherapy protocol which could lead to better therapy response.

Recellularized Colorectal Cancer Patient-Derived Scaffolds as In Vitro Pre-Clinical 3D Model for Drug Screening

Colorectal cancer (CRC) shows highly ineffective therapeutic management. An urgent unmet need is the random assignment to adjuvant chemotherapy of high-risk stage II and stage III CRC patients without any predictive factor of efficacy. In the field of drug discovery, a critical step is the preclinical evaluation of drug cytotoxicity, efficacy, and efficiency. We proposed a patient-derived 3D preclinical model for drug evaluation that could mimic in vitro the patient’s disease. Surgically resected CRC tissue and adjacent healthy colon mucosa were decellularized by a detergent-enzymatic treatment. Scaffolds were recellularized with HT29 and HCT116 cells. Qualitative and quantitative characterization of matched recellularized samples were evaluated through histology, immunofluorescences, scanning electron microscopy, and DNA amount quantification. A chemosensitivity test was performed using an increasing concentration of 5-fluorouracil (5FU). In vivo studies were carried out using zebrafish (Danio rerio) animal model. Permeability test and drug absorption were also determined. The decellularization protocol allowed the preservation of the original structure and ultrastructure. Five days after recellularization with HT29 and HCT116 cell lines, the 3D CRC model exhibited reduced sensitivity to 5FU treatments compared with conventional 2D cultures. Calculated the half maximal inhibitory concentration (IC50) for HT29 treated with 5FU resulted in 11.5 µM in 3D and 1.3 µM in 2D, and for HCT116, 9.87 µM in 3D and 1.7 µM in 2D. In xenograft experiments, HT29 extravasation was detected after 4 days post-injection, and we obtained a 5FU IC50 fully comparable to that observed in the 3D CRC model. Using confocal microscopy, we demonstrated that the drug diffused through the repopulated 3D CRC scaffolds and co-localized with the cell nuclei. The bioengineered CRC 3D model could be a reliable preclinical patient-specific platform to bridge the gap between in vitro and in vivo drug testing assays and provide effective cancer treatment.

Examination of Factors Influencing the Success Rates of an In Vitro Chemosensitivity Test for Lung Cancer Using Collagen Gel Droplet-Embedded Cultures

Objective: We evaluated the success rates and factors influencing the success rates of the Collagen Gel Droplet-Embedded Culture Drug Sensitivity Test (CD-DST) for pleural effusate in lung cancer patients with carcinomatous pleurisy.Materials and methods: Thirty-two patients with lung cancer having carcinomatous pleurisy were enrolled in this study between January 2014 and May 2017. Their pleural effusions were analysed by CD-DST, and the subjects were classified into two groups: successful cases and unsuccessful cases, based on whether the test was able to successfully determine drug sensitivity. We investigated whether the properties of the pleural effusate might influence the success rate of CD-DST. We observed that the CD-DST tended to have good success rates in patients with higher pH, higher percentage of lymphocytes, higher cell counts, and lower percentage of neutrophils in their pleural effusate. The red blood cell count was higher in the effusate of unsuccessful than successful cases.Conclusion: Our results suggest that the diagnostic success rates of CD-DST for pleural effusion from patients with lung cancer might improve by appropriate sample selection.