Quantitative assessment of nanoparticle uptake and trafficking in advanced 3D cell models: A high-content screening
1Organoid Core Facility, Gustave Roussy Institute, Villejuif, France; Department for BioMedical Research, University of Bern, Bern, Switzerland; Department of Urology, Inselspital, Bern University Hospital, Bern, Switzerland.
Abstract:
Nanoparticles (NPs), the most common physical form of drug or therapeutic delivery systems enhance treatment safety and efficacy thanks to protection and vehiculation of their payload into the targeted tissue. Recently, nanomedicines have received increasing interest in the area of cancer therapeutics. However, the failure to overcome biological barriers at the tissue and cellular levels prevents NP distribution and delivery, which carries unsuccess in clinical trials. With the introduction of advanced three-dimensional (3D) tumor replicas such as cancer spheroids and patient-derived organoids (PDOs) that provide a physiological 3D tissue-like context, the intrinsic NP-cell interaction features such as cellular uptake and intratumor behavior can be directly tested on a translational and preclinical basis. High-content screening (HCS) plays a pivotal role for predicting the efficacy of each NP design in an automated and unbiassed manner. This chapter presents in detail some protocols developed for confocal microscopy HCS on miniaturized cultures of tumor spheroids and PDOs, quantitative profiling of NP intra-spheroid transport and RNA-interference screening to unveil NP penetrance cellular effectors. In perspective, these modules put together, envision a core for a nano-PDO HCS workflow to advance cancer nanomedicine development.
