Fluorescence‐Guided Spatial Drug Screening in 3D Colorectal Cancer Spheroids

Abstract

The limited recapitulation of critical cancer features in two‐dimensional (2D) cultures causes poor translatability of preclinical results from in vitro assays to in vivo tumor models. This contributes to slow drug development with a low success rate. Three‐dimensional (3D) cultures better recapitulate the tumor microenvironment, enabling more accurate predictions when screening drug candidates and improving the development of chemotherapeutics. Platinum (Pt) (IV) compounds are promising prodrugs designed to reduce the severe systemic toxicity of widely‐used Food and Drug Administration (FDA)‐approved Pt(II) drugs such as cisplatin. Here, we present spatiotemporal evaluations in 3D colorectal cancer (CRC) spheroids of mitochondria‐targeting Pt(IV) complexes. CRC spheroids provide a greater pathophysiological recapitulation of in vivo tumors than 2D cultures by a marked upregulation of the ABCG2 chemoresistance marker expression. Furthermore, we introduce new 3D‐staining protocols to evaluate the real‐time decrease in mitochondria membrane potential (ΔΨ) in CRC spheroids, and a Pt‐sensing dye to quantify the Pt mitochondrial accumulation. Finally, we demonstrate a correlation between in vitro results and the efficacy of the compounds in vivo. Overall, the CRC spheroids represent a fast and cost‐effective model to assess the behavior of Pt compounds in vitro and predict their translational potential in CRC treatment.This article is protected by copyright. All rights reserved