Patient-Derived 3D Models for Preclinical Drug Response Evaluation in Neuroblastoma

Abstract

Abstract Background Neuroblastoma is the most common solid childhood tumor outside the central nervous system. Metastatic and relapsed disease in neuroblastoma patients still has a poor prognosis. New treatment methods to improve therapy and outcomes for patients with advanced neuroblastoma are urgently needed. However, clinical trials currently only allow for testing few substances in even fewer patients. This increases the need to improve and advance preclinical tumor models to preselect favorable candidates for novel therapeutics. Aims This study investigates the advantages and limitations of using bioreactor-based perfused 3D models in for preclinical drug testing in solid tumors exemplified in neuroblastoma. Methods We established perfused bioreactor-based 3D models using neuroblastoma cell lines and tissue slice-culture and evaluated cell viability. Several inhibitors against hypoxia-related factors, associated with tumor progression, were tested including Aqb011 against aquaporin 1 (AQP1) and SLC-0111 against carbonic-anhydrase IX (CAIX). Isothermal microcalorimetry was used for measuring real time thermogenesis in 3D cell and tissue constructs. Results 3D neuroblastoma cell constructs presented with similar morphological features compared to neuroblastoma cells in intact tumor tissue featuring small, round and blue cells. Perfused neuroblastoma slice-culture maintained its primary structure in the bioreactor for up to 10 days. Both 3D neuroblastoma cell constructs and neuroblastoma tissue responded to inhibition with significant decrease of thermogenesis measured by microcalorimetry, if the correlating hypoxic factor (AQP1 or CAXI) was expressed. The AQP1 inhibitor Aqb011 and the CAIX inhibitor SLC-0111 decrease heat production of 3D neuroblastoma cultures. Conclusions We newly establish two preclinical models for neuroblastoma cells and tissue in a 3D structure. Drug response monitoring can be applied in these models using isothermal microcalorimetry. Monitoring times can be increased by use of the perfused bioreactor. Our models are an important first step to preclinically evaluate new drugs using patient-derived tumor tissue exemplified for neuroblastoma.