ESR1: Omics technologies to understand the ER stress response
Studies in ER stress to date have largely used traditional single gene and protein centred approaches in diverse disease models, and there has been no comprehensive study to understand the global response to ER stress in normal cells.
ESR1 will use cutting edge “omic” technologies to comprehensively detect the global cellular changes in RNA, Protein, glycosylation and metabolites due to ER stress in normal epithelial cell lines.
ESR2: Contribution of RIDD activity to cell survival/death
To uncover key RIDD substrates that influence cell survival and cell death ESR2 will use recombinant IRE1α cytosolic domain and shRNA technology to assess the contribution of RIDD substrates to normal cellular processes and on cancer cell characteristics (e.g., chemoresistance and cell growth, adhesion/migration and invasion
ESR3: ER stress regulated autophagy and autophagic cell death
Autophagosome may function as a platform facilitating pro-caspase-8 activation and thereby influencing cell fate. ESR3 will use a proteomics approach to investigate the full components of the caspase-8 activating complex, including the inhibitors and regulators of the complex.
Genetic approaches (overexpression, siRNA, Crispr/CAS9) will be used to test the effect of different components of caspase-8 activating complex on autophagy and cell fate.
ESR4: Deciphering and exploiting aggresome-associated cell death initiation to predict anti-cancer treatment efficacy.
ESR4 will investigate the kinetics and quantities of apoptosis vs. necroptosis activation using semi-HTS flow cytometry and biochemistry in melanoma models.
Highly sensitive FRET-based time-lapse imaging of caspase activation in combination with biophotonic indicators for necroptotic membrane permeabilisation will provide insight into whether both cell death modes can be co-activated at the single cell level.
Components of systems biological network analysis will be employed for data interpretation and additional mechanistic insight, leading towards the development of predictive treatment response models.
ESR5: Functional role of the newly discovered PERK-interacting partners
Understanding the functional impact of the PERK-interactome and its dynamic changes during ER stress will be crucial to understand the global role of PERK in disease conditions (e.g., cancer) and to develop strategies aimed at blocking specific interactions, while sparing others.
The host laboratory has identified new molecular partners of PERK, which modulate the dynamic of interorganellar contact sites and response to Ca2+ fluxes, independent of the UPR.
ESR5 will investigate how specific perturbations/disruptions of the newly identified PERK-interactome (shRNA, CRrisp-CAS9, pharmacological inhibition) affect key cancer cell features under basal or ER stress conditions.