Infection of the human intestine by enteroviruses: A CRISPR-based screening approach

Informations projet

EvISC

N° de convention de subvention: 795904

DOI

10.3030/795904

Projet clôturé le 31 Août 2019

Date de signature de la CE 7 Mars 2018

Date de début 1 Janvier 2019

Date de fin 31 Decembre 2020

Financé au titre de

• EXCELLENT SCIENCE - Marie Skłodowska-Curie Actions

Coût total

€ 200 194,80

Contribution de l’UE

€ 200 194,80

Coordonné par

KOBENHAVNS UNIVERSITET
Denmark

Periodic Reporting for period 1 - EvISC (Infection of the human intestine by enteroviruses: A CRISPR-based screening approach)

Période du rapport: 2019-01-01 au 2020-12-31

Résumé du contexte et des objectifs généraux du projet

Managing infection diseases rely on the availability of medical treatment. While medication, including vaccines, have been developed and are successfully used for treating and protecting individuals, to date there are still many infections for which treatment options are limited or not available. Human Enterovirus (HEV) infections are of public health concern as globally millions of people become infected each year. Most HEV infections cause only mild symptoms, however severe pathologies can occur, especially in immunocompromised individuals as well as in infants, children and the elderly. To date, no enteroviral therapies exist and new strategies are urgently needed to counter HEV outbreaks and severe pathologies in populations at risk, in particular given the fact that many countries have stopped vaccinating against the most prominent member of the HEV family, polio.

This project aims at identifying host factors crucial for the HEV life cycle by combining for the first time, state-of-the-art high-throughput CRISPR technology with physiologically relevant primary cultures of human intestinal epithelial cells (organoids). The ultimate goal is to identify novel host factors that are key regulators of the viral life cycle and might pave the way to the discovery of novel drug targets for a broad-spectrum treatment against HEV infections. Two overall objectives have been defined for this project, which are (1) identifying novel host factors essential for the life cycle of human enteroviruses (HEVs) upon infection of human intestinal organoids, and (2) deciphering the roles of these identified host factors in promoting HEV infection in humans.

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

The main results obtained during this reporting period include hands-on training (two months secondment) on how to handle HEVs and to perform various virus-related assays. Furthermore, pilot experiments using HEVs and intestinal organoids revealed that these cells constitute a suitable model system to study HEV infections, and that most HEVs tested rapidly infect and within 2-3 days kill the cells.
Whole-genome CRISPR screens are usually carried out using immortalized cell lines that are easy to culture and can be grown in high numbers. In order to be able to carry out a CRISPR screen in intestinal organoids we turned towards a bioreactor culture system that supports growth of high numbers of organoids in a single vessel and allows for easy subculturing. In addition, this system can also be used directly for virus-related experiments.

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)

I am terminating the fellowship after 8 months and thus, have little to report in this section. In terms of impact, this project has had a positive effect on following aspects:
i) host lab; implementing a bioreactor as an organoid culture system in the host lab provides other lab members or even the institute with a novel tool
ii) collaboration; thanks to this project a collaboration between the host lab and the secondment lab was established
iii) outreach; a one-week biology course was held at a high-school in Switzerland and due to its success and positive feedback from teachers and students the course will be implemented again in the next school year

HEV (Coxackievirus A2) infected intestinal organoid. Virus in green and host cell nuclei in blue.