Culture and Drug Profiling of Patient Derived Malignant Pleural Effusions for Personalized Cancer Medicine

Abstract

IntroductionThe use of patients’ own cancer cells for in vitro selection of the most promising treatment is an attractive concept in personalized medicine. Human carcinoma cells from malignant pleural effusions (MPEs) are suited for this purpose since they have already adapted to the liquid environment in the patient and do not depend on a stromal cell compartment. Aim of this study was to develop a systematic approach for the in-vitro culture of MPEs to analyze the effect of chemotherapeutic as well as targeted drugs.MethodsMPEs from patients with solid tumors were selected for this study. After morphological and molecular characterization, they were cultured in medium supplemented with patient-derived sterile-filtered effusion supernatant. Growth characteristics were monitored in real-time using the xCELLigence system. MPEs were treated with a targeted therapeutic (erlotinib) according to the mutational status or chemotherapeutics based on the recommendation of the oncologists.ResultsWe have established a robust system for the ex-vivo culture of MPEs and the application of drug tests in-vitro. The use of an antibody based magnetic cell separation system for epithelial cells before culture allowed treatment of effusions with only moderate tumor cell proportion. Experiments using drugs and drug-combinations revealed dose-dependent and specific growth inhibitory effects of targeted drugs.ConclusionsWe developed a new approach for the ex-vivo culture of MPEs and the application of drug tests in-vitro using real-time measuring of cell growth, which precisely reproduced the effect of clinically established treatments by standard chemotherapy and targeted drugs. This sets the stage for future studies testing agents against specific targets from genomic profiling of metastatic tumor cells and multiple drug-combinations in a personalized manner.

In vitro cell culture of patient derived malignant pleural and peritoneal effusions for personalised drug screening

“…Therefore, for MSE cytologic diagnosis, Pan-CK is not be a proper marker for recognising tumour cells. Clearing of mesothelial cells using magnetic cell separation (MACS systems) might be crucial to establish pure cultures of MSE to be used for screening assays [13].…”

“…However, only modest gains have been made in long-term patient survival due to multidrug-resistant and highly aggressive characteristics of tumours growing in either pleural or peritoneal cavity. Screening of patients' own samples for in vitro chemo/immunotherapeutic agent selection enables the optimisation of individual therapeutic regimens [13]. To date, the utilisation of MSE 2D cell culture for drug screening and other experimental studies has already been demonstrated from literature evidences [8,13,14].…”

“…Screening of patients' own samples for in vitro chemo/immunotherapeutic agent selection enables the optimisation of individual therapeutic regimens [13]. To date, the utilisation of MSE 2D cell culture for drug screening and other experimental studies has already been demonstrated from literature evidences [8,13,14]. Moreover, in vitro MSE models generated either with scaffold-free techniques or with hydrogel matrix support were also used in few studies [15,16].…”

Management of cytological material, pre‐analytical procedures and bio‐banking in effusion cytopathology

“…As a recently established and rapidly expanding research field, biobanking also has an important role in ex vivo primary cell cultures of tumour cells derived from effusions. Viability of the frozen cells allows broad molecular characterisation and functional drug sensitivity/resistance profiling for multiple predictive markers and therapy targets …”

The methods of in vitro selection of chemotherapeutic druds for individual chemotherapy of maligant tumors in patinets

“…Система xCELLigence позволяет определить цитостатическое, цитотоксическое и митогенное воздействие таргетных препаратов (эрлотиниба) и химиопрепаратов (цисплатина), в режиме реального времени с 15 мин интервалом в течение 90 ч. Препараты добавляют через 1 сут к клеткам том 5 №3 / 2018 Онкология / Oncology аденокарциномы легкого человека, выделенных из злокачественной легочной эффузии. Это дает возможность построить динамические кривые и определить латентный период действия препарата на рост клеток [71].…”