Robotic system for assessing metabolic activity as a read-out for drug response of pediatric tumors

点击翻译按钮获取中文摘要

The Translation Program of the Hopp Children's Cancer Center Heidelberg (KiTZ) headed by the applicant, is comprising groups, which are working on better understanding and developing new treatment options for children with different pediatric cancer entities.Within our INFORM registry study (INdividualized Therapy FOr Relapsed Malignancies in Childhood) molecular analysis of more than 1000 tumor relapses proved that about 50% of the tumors harbour putatively druggable events. However, therapeutic targets with high evidence for response and clinical benefit are only detected in a minority of tumours. To identify more therapeutic options for pediatric cancer patients, we therefore aim to integrate functional single and combination drug response data with our well established INFORM molecular profiling analysis (Low Coverage Whole Genome Sequencing, Whole Exome Sequencing, RNA Sequencing, DNA Methylation). To develop the drug screening platform towards future companion diagnostics and improvement of clinical success rates we follow different strategies to improve recapitulation of in vivo disease and therapy. Amongst others, these include 1) focusing on approved compounds, 2) three-dimensional tumor cell cultures (spheroid cultures) in 384-well format, 3) standardized protocols for sample preparation and screening, and 4) quality controls for the assay read-out. We are currently assessing drug response by image-based analyses on a high-content microscope with a combination of fluorescent probes and with consideration of state-of-the art HCI assay development and analysis guidelines.However, according to EU regulations (2017/746 (IVDR)) for in vitro diagnostics, functional drug response data have to be validated by a second, technically independent, method to allow translation into potential clinical phase I/II studies. We therefore apply for an automated system to measure metabolic activity, as an equivalent to residual cell viability, of the three-dimensional tumor spheroids after drug exposure. Moreover, this robotic system will allow us to increase the throughput of other drug screening projects of the group and in collaboration with other KiTZ members.