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Dynamic regulation of paracellular channel gating

Project description

Defining tight junction gating kinetics

In epithelial layers, cells are joined through tight junctions, multiprotein complexes that regulate the selective transport between organ compartments. Claudin proteins play a fundamental role in these structures, forming highly selective conductance channels. Mutations in claudin genes cause hypomagnesaemia, renal failure, and other human diseases, underscoring the clinical importance of these proteins. The EU-funded DynChan project will develop an innovative technology to study the mechanism underlying transport regulation through claudin channels. A nanopillar array chip will employ nanoelectrodes to probe single-channel events over many tight junctions and define claudin pore function. Results will accelerate our understanding of tight junction biology and contribute to the design of novel therapeutic interventions.

Objective

Epithelial paracellular, i.e. tight junction, permeability is largely defined by the integrated functions of claudin proteins that can either seal the paracellular space or form highly-selective conductance channels. The importance of claudins is exemplified by the many human diseases caused by barrier dysregulation and claudin mutations.
The host laboratory recently reported the first measurements of single channel tight junction currents, thereby demonstrating that claudin channels transition between open and closed states. The central hypothesis of this application is that claudin channel activity is regulated by specific molecular interactions.
Unfortunately, the trans-tight junction patch-clamp method developed by the host laboratory is extremely labor intensive and unable to capture more than a small section of a single tight junction, making it unsuitable for comprehensive analyses. To overcome this obstacle, we first aim to develop a nanopillar array chip that will supersede the patch-clamp method. Cells grown over and around the nanopillars will form tight junctions above the nanoelectrode at the tip of each nanopillar. This will make it possible to measure large numbers of single-channel events over many junctions.
The second aim will exploit the nanopillar chip to define the conductances and gating activities of claudin proteins and the mechanisms by which they are regulated. This novel technology will also allow others to analyze claudin function in health and disease. The nanopillar chip and data generated using this tool will accelerate our understanding of tight junction biology and enable development of channel modulators that, in a manner analogous to the advances enabled by transmembrane ion channel modulators, will lead to novel therapeutic approaches.

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MSCA-IF - Marie Skłodowska-Curie Individual Fellowships (IF)

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Call for proposal

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

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Coordinator

AGENCIA ESTATAL CONSEJO SUPERIOR DE INVESTIGACIONES CIENTIFICAS
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.

€ 245 732,16
Address
CALLE SERRANO 117
28006 MADRID
Spain

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Region
Comunidad de Madrid Comunidad de Madrid Madrid
Activity type
Research Organisations
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Total cost

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€ 245 732,16

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