Integrated 3D Light-Sheet and 2D Multiplex Imaging for Deep Histological Profiling of a Somatic Mouse Glioblastoma Model

Abstract

Glioblastoma is a devastating brain cancer. Despite intense research, patient survival has not significantly increased over the past decades and efficient treatment is currently not available. Therefore, the fundamental understanding of the disease, based on the development of relevant animal models, combined with the development of efficient tools for their deep analysis, represents a priority. Neural Stem cells in the subventricular zone of the forebrain have been identified as cells of origin for glioblastoma, leading to the development of new somatic lineage models based on in vivo brain electroporation. While such models have been characterized in depths by sequencing approaches, systematic histological analyses are currently scarce. Here we present the multimodal histological characterization of a transgenesis independent somatic glioblastoma model in mice. Using 3D light sheet imaging we demonstrate that the model is highly reproducible, allowing quantitative evaluation of tumor growth over large cohorts. Using multiplex imaging by MICS technology we systematically characterize the cellular landscape and molecular composition of the induced tumors, as well as their micro- and macro-environments, and provide a resource of mouse compatible antibodies for cancer research. Finally, we use the model to show that tissue clearing and 3D light sheet microscopy of whole brains can be combined with subsequent multiplex imaging, allowing deep spatial characterization of the tumor proteome in pre-identified brain regions.