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Predicting Apoptosis Engagement Downstream of Caspases in vivo

Project description

Understanding predictive parameters of apoptosis downstream of caspase activation

Epithelial cell death is part of the developmental process and tissue homeostasis. Recent studies using the Drosophila pupal notum (a single-layer epithelium) demonstrated that cells engage in apoptosis at variable levels of effector caspase activity. This suggests a new layer of regulation of cell death with important physiological roles. The ERC-funded PrApEDoC project proposes to combine quantitative live imaging, quantitative optogenetic perturbations, and supervised machine learning to define predictive apoptosis parameters downstream of caspase activation. Next, the project will use spatial omics technologies combined with an optogenetic screening to identify factors modulating sensitivity to caspases. The goal is to uncover new physiological functions of caspase sensitivity modulation and characterise its impact on tissue morphogenesis and oncogenic cell elimination.

Objective

Epithelial cell death is widespread during development and tissue homeostasis. While the core pathway of apoptosis is well known, we lack predictive power to understand when and where a specific cell will die in a tissue. Yet, this should have important consequences for clone selection and tissue homeostasis, as exemplified by the process of cell competition. This context dependent cell elimination is essential for tissue homeostasis and tumour progression, but we know very little about its prevalence during normal development. Recent works have characterised multiple non-apoptotic functions of effector caspases and a high frequency of cells undergoing transient caspase activation without death. Yet, what distinguishes a dying from a surviving cell downstream of effector caspase activation is completely unknown. Using quantitative live imaging of effector caspase sensors and caspase activity tuning by optogenetics in the Drosophila pupal notum (a single layer epithelium), we found that cells engage in apoptosis at very variable levels of effector caspase activity. This suggested that the engagement in apoptosis can be developmentally modulated downstream of the most terminal caspase activation and that a completely new layer of regulation of cell death could have important physiological roles. Here, we propose to combine quantitative live imaging, multivariate model, supervised machine learning and quantitative perturbations through optogenetics to define parameters predictive of the engagement in apoptosis downstream of caspase activation. We will then use spatial omics combined with an optogenetic based screening to identify new factors modulating the sensitivity to caspases. Eventually we will unravel the physiological functions of caspase sensitivity modulation by defining a new paradigm of physiological cell competition and clone selection, and characterising its impact on tissue morphogenesis and oncogenic cell elimination.

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Host institution

INSTITUT PASTEUR
Net EU contribution
€ 1 999 625,00
Address
RUE DU DOCTEUR ROUX 25-28
75724 Paris
France

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Region
Ile-de-France Ile-de-France Hauts-de-Seine
Activity type
Research Organisations
Links
Total cost
€ 1 999 625,00

Beneficiaries (1)