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On the development of novel molecular and microscopy methodologies for the interrogation of host-pathogen interactions at the single-cell level

Project description

Delineating the host–pathogen war

Host–pathogen interaction has led to the evolution of strategies from both sides to exert their dominance. Delineation of this interplay is central for the design and development of effective methods to eradicate pathogens. Scientists of the EU-funded FISHnCRISPS project will use Salmonella typhi as a model to perform genetic screening using CRISPR interference. The idea is to isolate various virulence factors and determine their impact on host responses. In addition, the simultaneous transcriptome profiling of host and pathogen at the single cell level will help scientists understand the heterogeneity in outcome often observed among individuals battling with the same pathogen.

Objective

Over the course of host-pathogen evolution, both the players in this game have set out strategies to overcome each other, by adapting to challenging environments and camouflaging on the one side, and by detecting and eradicating the enemy on the other. In this context, the identification of the pathogenic potential, that is, the genetic elements conferring virulence and how they subvert the host’s defences, is fundamental. With a multiplexed approach, I aim at developing a pooled genetic screen using CRISPR interference to produce pathogen knocked-down libraries impaired in a curated subset of virulence factors of Salmonella Typhi. This pathogen is of particular interest because of the severity of its systemic infection combined with its human host-restriction, which left research lagging behind. Several cellular systems (macrophages, PBMCs, organoids) will be probed upon infection of S. typhi mutants, and the host transcriptome retrieved by single-cell RNA-seq. This analysis will provide a systematic and thorough map of the cause and effect network of the pathogen virulence factors and host responses.
Interestingly, in this pathogen-host “war”, each battle can end with a different winner, even when the teams are comprised of the same players, meaning that the heterogeneity in the cellular state of both the pathogen and the host previous to the infection, can lead to the occurrence of diverse outcomes within a single population. The pathogen can either replicate, persist or be cleared by the host. This intrinsic cell-to-cell variability posits the importance of profiling simultaneously the host and the pathogen at the level of single cells. To achieve this goal, I will profile the transcriptomes of the host and the pathogen at the same time, by using multiplexed smFISH. This analysis will quantitatively untangle the contribution of heterogeneity to disease outcomes.

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Funding Scheme

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MSCA-IF-EF-ST - Standard EF

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Call for proposal

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(opens in new window) H2020-MSCA-IF-2019

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Coordinator

WEIZMANN INSTITUTE OF SCIENCE
Net EU contribution

Net EU financial contribution. The sum of money that the participant receives, deducted by the EU contribution to its linked third party. It considers the distribution of the EU financial contribution between direct beneficiaries of the project and other types of participants, like third-party participants.

€ 173 464,32
Address
HERZL STREET 234
7610001 Rehovot
Israel

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Activity type
Higher or Secondary Education Establishments
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Total cost

The total costs incurred by this organisation to participate in the project, including direct and indirect costs. This amount is a subset of the overall project budget.

€ 173 464,32
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