A humanized breast cancer microorganoid platform for the ranked evaluation of tumor microenvironment-targeted drugs by light sheet fluorescence microscopy

Abstract

Abstract Targeting the supportive tumor microenvironment (TME) is an approach of high interest in cancer drug development. However, assessing TME-targeted drug candidates presents a unique set of challenges: Adequate assays need to recreate the TME at least in part and provide intricate information about drug-induced changes in the TME’s interactions. We have developed a comprehensive screening platform that allows to monitor, quantify, and rank drug-induced effects in self-organizing, vascularized tumor microorganoids (TMOs). Fully humanized, the confrontation of four different cell populations makes it possible to study complex changes in composition and cell-cell interaction. The platform is highly modular, allowing for adjustments regarding tumor entity, TME composition, or for genetic manipulation of individual cell populations. Treatment effects are recorded by light sheet fluorescence microscopy and translated by an advanced image analysis routine in processable multi-parametric datasets. The detailed data output allows for handling a wide range of potential inquiries. Nevertheless, the system proved to be robust, with strong interassay reliability. We demonstrate the platform's utility for the side-by-side evaluation of TME-targeted antifibrotic and antiangiogenic drugs. The platform's output delivered a broad scope of information about treatment effects, enabling clear distinction of even closely related drug candidates according to projected therapeutic needs. Moreover, the modular character allowed for the differential evaluation of genetically targeting different cellular components, adding new possibilities for tailoring selective drugs.