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

由申请人领导的海德堡霍普儿童癌症中心（KiTZ）的翻译项目由小组组成，这些机构致力于更好地了解和开发针对不同儿童癌症实体儿童的新治疗方案。在我们的INFORM登记研究中，（儿童复发性恶性肿瘤个体化治疗）对1000多例肿瘤复发的分子分析证明，约50%的的肿瘤中含有可注射的药物。然而，具有高度反应和临床益处证据的治疗靶点仅在少数肿瘤中检测到。为了为儿童癌症患者确定更多的治疗选择，我们的目标是将功能性单一和联合药物反应数据与我们完善的INFORM分子分析（低覆盖率全基因组测序，全外显子组测序，RNA测序，DNA甲基化）相结合。为了开发面向未来伴随诊断和提高临床成功率的药物筛选平台，我们遵循不同的策略来改善体内疾病和治疗的重演。其中，这些包括1）专注于批准的化合物，2）384孔格式的三维肿瘤细胞培养物（球状体培养物），3）用于样品制备和筛选的标准化方案，以及4）用于测定读出的质量控制。我们目前正在通过结合荧光探针在高容量显微镜上进行基于图像的分析来评估药物反应，并考虑到最先进的HCI测定开发和分析指南。（2017/746（IVDR））对于体外诊断，功能性药物反应数据必须由第二个技术上独立的，方法，以允许转化为潜在的临床I/II期研究。因此，我们申请了一种自动化系统来测量药物暴露后三维肿瘤球体的代谢活性，相当于残留细胞活力。此外，这个机器人系统将使我们能够增加该集团其他药物筛选项目的吞吐量，并与其他KiTZ成员合作。

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.