Defining heterocellular signalling within the intratumoral stem cell niche of colorectal cancer

Projektinformationen

IntratumoralNiche

ID Finanzhilfevereinbarung: 803608

DOI

10.3030/803608

Projekt abgeschlossen

EK-Unterschriftsdatum 25 Oktober 2018

Startdatum 1 Januar 2019

Enddatum 31 Dezember 2023

Finanziert unter

• EXCELLENT SCIENCE - European Research Council (ERC)

Gesamtkosten

€ 1 500 000,00

EU-Beitrag

€ 1 500 000,00

Koordiniert durch

UNIVERSITAIR MEDISCH CENTRUM UTRECHT
Netherlands

Periodic Reporting for period 4 - IntratumoralNiche (Defining heterocellular signalling within the intratumoral stem cell niche of colorectal cancer)

Berichtszeitraum: 2023-07-01 bis 2023-12-31

Zusammenfassung vom Kontext und den Gesamtzielen des Projekts

Purpose:
Cells in a tumor are highly heterogeneous. The role and consequence of having multiple cell types
within a cancer is mostly centered towards the function of cancer stem cells (CSCs) since they are the driving
forces of tumor growth. However, even the most malignant cancers contain differentiated tumor cells. Is there a functional role for these cells,
for instance by providing signaling cues that support CSC function. In other words, do colorectal tumors build their own stem cell supporting niche?

Importance for society:
My overarching aim is to identify and characterize the signaling pathways in cancer stem cells and differentiated tumor cells (tumor-intrinsic niche) that are vital for colorectal tumor growth and metastasis formation.

Objectives:
-Identify signaling pathways that act between cancer stem cells and differentiated tumor cells
-Develop new technological strategies to characterize cellular behavior, cell cycle kinetics, and signaling activities in real-time and with single-cell resolution in patient organoids
-Functional investigation of candidate genes that are essential components of the intratumoral stem cell niche

Arbeit, die ab Beginn des Projekts bis zum Ende des durch den Bericht erfassten Berichtszeitraums geleistet wurde, und die wichtigsten bis dahin erzielten Ergebnisse

We characterized the cell type dynamics, composition, and transcriptome of early colorectal cancer liver metastases. This revealed that failure to establish cellular heterogeneity (i.e. presence of cancer stem cells and differentiated tumor cells) through YAP-controlled epithelial self-organization prohibits the outgrowth of micrometastases (Heinz et al., Cancer Res. 2022).

We have developed a new CRISPR pipeline that allows for fast, highly efficient, and error-free fluorescent genetic knock-ins in human organoids, both normal and cancer. The method is published and the palette of targeting vectors are publicly available from Addgene (Bollen, Hageman, et al., PLOS Biol, 2022).

We studied cell cycle kinetics in patient tumor organoids to map chromosomal instability in colorectal cancer via live-cell imaging of 3D organoids. We demonstrate a) that CIN is widespread in these cancers regardless of mutational landscape and b) that the combination of CIN levels and tolerance for mitotic errors shape karyotype diversification and heterogeneity landscapes. (Bolhaquiero, Ponsioen et al., Nat Genet 2019).

Regarding real-time analysis of signaling activity, we report the most optimal FRET biosensor to measure ERK activity in single-cells in real-time in patient organoids. Second, we developed strategies to measure quantitative signaling activity in 3D patient organoids. Third, we document how EGFR amplifies downstream ERK activity in human CRC, even in the presence of mutant KRAS or BRAF. The work explains EGFR as a successful drug target in widely applied combination therapies against CRC. (Ponsioen et al., Nat Cell Biol 2021).

Currently our work in ongoing to continue innovating our methodologies to assess single-cell signaling activities in real-time, and we work with sophisticated genetic knock-ins to address signaling function of diverse cell types in human tumors.

Fortschritte, die über den aktuellen Stand der Technik hinausgehen und voraussichtliche potenzielle Auswirkungen (einschließlich der bis dato erzielten sozioökonomischen Auswirkungen und weiter gefassten gesellschaftlichen Auswirkungen des Projekts)

Developed strategies that provide insights into real-time signaling activity in tumor organoids and its quantitative assessment upon targeted therapies is a technological breakthrough. We aim to expand our toolbox to study more signaling pathways, and innovate the strategy itself to perform long-term drug response analyses to incorporate the emergence of drug resistance.

patient organoids