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Hopp儿童癌症中心（KiTZ）的翻译项目由小组组成，这些小组正在努力更好地了解和开发患有不同儿科癌症实体的儿童的新治疗方案。在我们的INFORM注册研究（儿童复发恶性肿瘤的个体化治疗）中，对1000多例肿瘤复发的分子分析证明，约50%的肿瘤含有推定的药物事件。然而，具有高疗效和临床益处的治疗靶点仅在少数肿瘤中检测到。因此，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。为了开发未来伴随诊断和提高临床成功率的药物筛选平台，我们采用不同的策略来改善体内疾病和治疗的再现。其中包括1)专注于批准的化合物，2)384孔格式的三维肿瘤细胞培养（球形培养），3)样品制备和筛选的标准化方案，以及4)测定读数的质量控制。我们目前正在高含量显微镜上结合荧光探针进行基于图像的分析，并考虑到最先进的HCI检测发展和分析指南，以评估药物反应。然而，根据欧盟体外诊断法规（2017/746 (IVDR)），功能性药物反应数据必须通过第二种技术上独立的方法进行验证，才能转化为潜在的临床I/II期研究。因此，我们申请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，作为等效的残余细胞活力。此外，这个机器人系统将使我们能够提高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.