Genome-wide CRISPR/Cas9 screen in intestinal organoids for identification and molecular characterization of therapeutic targets to enhance tissue regeneration

Informations projet

ScrIntOR

N° de convention de subvention: 101026109

DOI

10.3030/101026109

Projet clôturé le 30 Septembre 2021

Date de signature de la CE 19 Mars 2021

Date de début 1 Mai 2021

Date de fin 30 Avril 2023

Financé au titre de

• EXCELLENT SCIENCE - Marie Skłodowska-Curie Actions

Coût total

€ 207 312,00

Contribution de l’UE

€ 207 312,00

Coordonné par

KOBENHAVNS UNIVERSITET
Denmark

Periodic Reporting for period 1 - ScrIntOR (Genome-wide CRISPR/Cas9 screen in intestinal organoids for identification and molecular characterization of therapeutic targets to enhance tissue regeneration)

Période du rapport: 2021-05-01 au 2023-04-30

Résumé du contexte et des objectifs généraux du projet

Around 3 million people in Europe are affected by inflammatory bowel disease (IBD), a devastating condition characterized by chronic inflammation of the gastrointestinal tract which causes severe damage to the intestinal epithelium. Current standard treatment of IBD focuses predominantly on reducing the inflammatory burden of the intestine, a symptomatic medication without promoting the regeneration of the damaged intestinal epithelium. This is because regeneration of this tissue is still very poorly understood. It is therefore of great importance to identify the molecular mechanisms driving epithelial regeneration under ulcerative conditions and thus facilitate the development of novel pharmaceutical compounds to improve tissue recovery in IBD patients.
In this project, we will combine in vitro intestinal organoid cultures with a CRISPR/Cas9 screening approach to (i) identify novel regulators of regeneration in the intestinal epithelium and (ii) decipher how this process is controlled at the molecular level. The outcome of the screen and the downstream validation will substantially enhance the current understanding of intestinal repair upon tissue damage and is likely to aid in the identification of molecular pathways that will constitute new therapeutic targets for patients suffering from IBD.

Travail effectué depuis le début du projet jusqu’à la fin de la période considérée dans le rapport et principaux résultats atteints jusqu’à présent

I have decided to terminate my MSCA fellowship after 5 months, as I have been awarded a postdoc fellowship from the Lundbeck Foundation, which will expand my funding period as a postdoc for another 3 years.
Within the reporting period, I have been successfully working on the characterization of intestinal organoid cultures as well as establishing the set-up for the genome-wide CRISPR/Cas9 screen, and I will be soon ready to perform the actual screen. I have therefore established intestinal organoid cultures under conventional conditions as well as grown in a bioreactor system, which will allow simultaneous growth of millions of organoids. These cultures upon appropriate stimulation can enter a regenerative state. In addition, I have generated intestinal epithelial cell lines which express Cas9 in an inducible manner.
Due to early termination of the MSCA fellowship, no exploitation or dissemination of the results was planned at this stage yet.

Progrès au-delà de l’état des connaissances et impact potentiel prévu (y compris l’impact socio-économique et les conséquences sociétales plus larges du projet jusqu’à présent)

As I terminated the fellowship at an early stage, the full impact of the project as well as the potential use of the project outcome will only be visible at a later point.
Nevertheless, the project combines state-of-the-art organoid cultures with a genome-wide CRISPR/Cas9 screening approach intending to identify the molecular drivers of intestinal regeneration. The results of the screen will be already a significant resource on their own, however, we are aiming to move beyond a purely descriptive approach by dissecting the molecular events that orchestrate tissue repair and validating critical pathway components in a colitis mouse model. Thus, this study will potentially facilitate the identification of novel drug targets for patients suffering from IBD.

Intestinal organoid expressing Cas9. Nuclei in blue, Cas9 in red, F-actin in cyan. Scale bar 100μm.