ROCKETS - a novel one-for-all toolbox for light sheet microscopy in drug discovery

Area of Science:

• Immunology
• Biomedical Imaging
• Preclinical Research

Background:

• Advancing immunotherapies requires better preclinical tools for target, biodistribution, safety, and efficacy assessment.
• Light sheet fluorescence microscopy (LSFM) provides fast, high-resolution imaging of large tissues but faces limitations due to laborious processing.
• Standardized protocols are needed to increase throughput and broaden LSFM applications in immunology.

Purpose of the Study:

• To develop a harmonized protocol for processing, clearing, and imaging entire mouse bodies for enhanced preclinical research.
• To apply this protocol with LSFM to study the biodistribution of an antibody targeting Epithelial Cell Adhesion Molecule (EpCAM).
• To identify novel EpCAM expression sites and assess their relevance for EpCAM-targeting immunotherapies.

Main Methods:

• Development of the Rapid Optical Clearing Kit for Enhanced Tissue Scanning (ROCKETS) protocol.
• Application of ROCKETS with LSFM for 3D imaging of whole mouse organs and bodies.
• Quantitative analysis of EpCAM distribution in mouse tissues and confirmation in human samples.

Main Results:

• ROCKETS enables comprehensive, high-resolution 3D imaging of entire mouse bodies.
• New EpCAM binding sites were identified, including gustatory papillae (tongue), choroid plexi (brain), and duodenal papillae.
• High EpCAM expression was confirmed in human tongue and duodenal tissues, highlighting potential clinical relevance.

Conclusions:

• ROCKETS combined with LSFM offers a standardized platform for preclinical evaluation of immunotherapies.
• The protocol facilitates quantitative co-localization studies within the microanatomical context of organs or whole organisms.
• Discovery of novel EpCAM sites has significant implications for the clinical translation of EpCAM-targeting immunotherapies.