Category: CEMM – Molecular Oncology

Abstract

Intestinal organoid cultures are a powerful tool to study epithelial cells in vitro, as they are able to proliferate and differentiate into all cell lineages observed in vivo. Co-culturing organoids with distinct genetic backgrounds provides an excellent approach to study contact dependent and independent interactions between healthy and mutant epithelial intestinal cells. Here, we provide 2D and 3D approaches to mouse organoid co-cultures using fluorescently labeled organoids and demonstrate the analysis of these co-cultures using flow cytometry and microscopy-based approaches.

For complete details on the use and execution of this profile, please refer to van Neerven et al., 2021.

Authors

Sanne M. van Neerven, Rana Ramadan, Milou S. van Driel, David J. Huels, Louis Vermeulen

Link

https://doi.org/10.1016/j.xpro.2021.101050

Abstract

The majority of colorectal cancers (CRCs) present with early mutations in tumor suppressor gene APC. APC mutations result in oncogenic activation of the Wnt pathway, which is associated with hyperproliferation, cytoskeletal remodeling, and a global increase in mRNA translation. To compensate for the increased biosynthetic demand, cancer cells critically depend on protein chaperones to maintain proteostasis, although their function in CRC remains largely unexplored. In order to investigate the role of molecular chaperones in driving CRC initiation, we captured the transcriptomic profiles of murine wild type and Apc-mutant organoids during active transformation. We discovered a strong transcriptional upregulation of Hspb1, which encodes small heat shock protein 25 (HSP25). We reveal an indispensable role for HSP25 in facilitating Apc-driven transformation, using both in vitro organoid cultures and mouse models, and demonstrate that chemical inhibition of HSP25 using brivudine reduces the development of premalignant adenomas. These findings uncover a hitherto unknown vulnerability in intestinal transformation that could be exploited for the development of chemopreventive strategies in high-risk individuals.

Authors

Sanne M van Neerven, Wouter L Smit, Milou S van Driel, Vaishali Kakkar, Nina E de Groot, Lisanne E Nijman, Clara C Elbers, Nicolas Léveillé, Jarom Heijmans, Louis Vermeulen

Link

https://doi.org/10.15252/emmm.202216194