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Functional analysis of the kinome and phosphatome as determinants of integrin phosphorylation in cancer

Project description

Integrin phosphorylation: a new cancer biomarker?

Integrins are a large family of proteins that attach the cell cytoskeleton to the extracellular matrix, serving as an adhesion point as well as a signal transducer. Altered integrin expression is frequently observed in cancer, where integrins seem to support oncogenic growth, migration and invasion. The EU-funded INTEGRIN REGULATION project is interested to delineate the activation of integrin beta 1 in breast cancer. Researchers will initially screen phosphorylated integrin beta 1 molecules and subsequently the enzymes responsible for this activation. Alongside information on the interacting partners of integrin beta 1, results will unveil novel mechanistic insight into breast cancer pathogenesis.

Objective

Integrins are key signalling molecules that mediate the complex relationship between cancer cells and their associated ECM; as such they have been implicated in every step of cancer progression. Their cytoplasmic domains are essential hubs for protein-protein interactions, where phosphorylation of key sites modulates integrin activity and oncogenic signalling. However, the regulators of integrin phosphorylation remain elusive. The proposed work will address the role of integrin beta1 in breast cancer epithelial and stromal cells, where increased expression and/or activity are correlated with poorer survival. To this end, the fellow has developed a Förster resonance energy transfer (FRET) biosensor for integrin beta1 phosphorylation, which will be applied in a kinome-/phosphatome-wide screen for regulatory kinases and/or phosphatases. This screening will be completmented with global mass spectrometry to identify phosphorylation-sensitive interactions with the integrin beta1 cytoplasmic domain. Out of these screens, the fellow will validate the identified kinases and/or phosphatases using three-dimensional functional assays, as well as standard biochemical approaches, in conjunction with live super-resolution imaging to pinpoint the subcellular localization of the interacting complex. The clinical relevance of the identified kinases and/or phosphatases will then be evaluated with immunohistochemistry staining of large breast cancer cohorts, along with functional validation using patient material from clinical collaborators. These aspects will provide an essential link from the mechanistic insights to improving patient outcomes from this work.

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Topic(s)

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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-2018

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Coordinator

TURUN YLIOPISTO
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.

€ 190 680,96
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.

€ 190 680,96
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