Morpho-molecular Functional Drug Response Analysis of Patient-Derived Organoids of Colorectal Cancer

Nanoparticle-based drug delivery systems have shown potential for cancer treatment. Due to their size, nanoparticles penetrate into tumors, facilitate drug uptake, enable efficient drug delivery and ensure efficacy at the target site. They present good pharmacokinetics, precisely target tumor cells, reduce side effects and drug resistance. Recently, patientderived organoids (PDOs) of colorectal cancer (CRC) patients proved to be a valuable tool to study the pathological, physiological, and genetic characteristics of CRC. In particular, they can be employed to test drugs in-vitro in a complex heterogeneous environment. We use UiO-66-NH2 nanoparticles as drug delivery carriers for 5- Fluorouracil (5-FU) to PDOs of CRC to study interactions between organoids and nanoparticles. Since PDOs preserve tumor heterogeneity to a significant extent, we investigate the structural, metabolic, and molecular changes in PDOs across different CRC and the impact of UiO-66-NH2 as carrier for 5-FU with our custom multimodal optical microscope offering complementary coregistered contrast from Optical Coherence Tomography (OCT), Multiphoton Microscopy (MPM), and Raman Microscopy (RM). We examine the interactions between nanoparticles and PDOs of CRC as well as their response to 5FU. We assess the influence of the drug by extracting morphological features from OCT/MPM images, distinguish PDOs phenotypic differences through RM molecular analysis, identifying alterations in relevant biomarkers such as lipids, proteins, collagen, DNA, further analyze molecular metabolic differences and conduct sample classification. We demonstrate the significant potential of our multimodal imaging system in the analysis of PDOs and nano-medicine effects, enabling high- precision morpho-molecular metabolic analysis and classification.