H2020Индивидуална стипендия2021–2024

DynChan · Dynamic regulation of paracellular channel gating

„Хоризонт 2020“ — Действия „Мария Склодовска-Кюри“

Период: 2021-03-16 → 2024-03-15
Финансиране от ЕС: 245 732 €
Участници: 2
Схема: MSCA-IF

Линиите свързват координатора с партньорите.

Накратко на български

Белтъците клаудини регулират преминаването на вещества между клетките, като техните мутации причиняват заболявания като фамилна хипомагнеземия. Разбирането на работата им чрез нови нанотехнологии помага за изясняване на механизмите на клетъчната пропускливост и търсенето на нови терапевтични подходи.

Този кратък обзор е генериран от изкуствен интелект

Кратко обяснение, генерирано от езиков модел по текста на CORDIS. Оригиналът е по-долу.

Резултати накратко

Dynamic regulation of paracellular channel gating

Tight junction proteins seal the space between adjacent epithelial and endothelial cells and form paracellular pores that regulate the paracellular transport: pore-forming claudins. Unfortunately, biophysics of these pore-forming claudins remains incompletely undefined. Mutations in pore-forming claudins cause human disease, including familial hypomagnesemia and hypercalcuria, neonatal sclerosing cholangitis associated with ichthyosis, and others. Underlying structure-function relationships will fundamentally advance our understanding of paracellular permeability and may, ultimately, lead to novel therapeutic approaches. The Dynchan project aims to develop a new technology that will enable high resolution analyses of claudin channel activity and regulation. The technology based on silicon chips with array of nanopillar electrodes is expected to provide the tool needed to develop foundational understanding of claudin biology. During the Dynchan project, we established conditions for cultivating functional, mature, properly assembled epithelial monolayers on high aspect ratio pillars localized to lateral intercellular spaces. We found that the pillar size strongly affected the assembling of tight junctions. For MDCK I cells, pillar structures of 1 µm in diameter and approximately up to 6 µm in height facilitated the formation of tight junctions in close proximity with the tips of the pillar structures. Additionally, through the application of micro and nanotechnologies, we have successfully fabricated miniature electrodes atop these pillars. This breakthrough allows us to record electrical currents crossing the tight junctions with an very low spatial resolution in the one micron range.

Текст от CORDIS, на английски · Данни: CORDIS, © Европейски съюз

Цел на проекта

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.

Оригинален текст от CORDIS (на английски).

Участници

AGENCIA ESTATAL CONSEJO SUPERIOR DE INVESTIGACIONES CIENTIFICAS · MadridКоординаторИспания
BRIGHAM INC · BostonСъединени щати