H2020Индивидуална стипендия2019–2021

hsPCF-FRET · Real-time characterisation of neuropeptide binding to a membrane receptor involved in pain and ischemic stroke

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

Период: 2019-04-01 → 2021-03-31
Финансиране от ЕС: 207 312 €
Участници: 1
Схема: MSCA-IF-EF-ST

Линиите свързват координатора с партньорите.

Накратко на български

Взаимодействието между протеина ASIC1a и пептида BigDynorphin се анализира чрез нови методи за наблюдение в реално време. Разбирането на този процес помага за разработването на лекарства срещу хронична болка и намаляване на уврежданията при исхемичен инсулт.

Този кратък обзор е генериран от изкуствен интелект

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

Резултати накратко

Real-time characterisation of neuropeptide binding to a membrane receptor involved in pain and ischemic stroke

One way for neurons to communicate with each other is through highly localised changes in brain tissue pH that activate acid-sensing ion channels (ASIC1a). The subtype ASIC1a plays a significant role in detecting low pH and makes essential contributions to learning and memory. However, some of the most prevalent neurological disorders, such as depression, chronic pain, and ischemia, also exhibit brain tissue acidification. Chronic pain alone has a prevalence of 19% in the adult European population, and ischemic stroke imposes a heavy socio-economic burden with currently extremely limited therapeutic options. One determining factor between physiological and pathological activation of ASIC1a appears to be the presence of BigDynorphin (BigDyn), a neuropeptide with physiological roles in circadian rhythm and appetite control. During neurological stress, however, BigDyn levels increase and, together with low tissue pH, activate ASIC1a to an extent that can be toxic to neurons, worsening the outcome of the related disorder. Targeting the dynorphin–ASIC1a interaction has therefore great therapeutic potential and could present an avenue to design a new generation of ASIC-selective drugs that could treat pain without the typical downsides of opioids or limit neurotoxicity during ischemic strokes. However, its exploitation was hampered by the limited mechanistic insight of the interaction and the lack of methods to directly assess it in detail. Thus, the project first aimed to establish a protocol using a fluorescence-coupled electrophysiology approach that would allow us to look at the interaction in a cell-based system. In a second step, we then wanted to identify the binding site of dynorphins on ASIC1a and analyse the binding processes under pathological conditions. We anticipate that the information gained from this project will aid the future design of ASIC1a inhibitors with the potential to treat chronic pain and ischemia.

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

Цел на проекта

Under physiological conditions, localized acidification of brain tissue serves as neuronal signal that get synaptically transduced via acid-sensing ion channels (ASIC1a). Local acidosis has, however, also been linked to some of the most prevalent neurological disorders such as chronic pain, ischemic stroke and psychiatric diseases. ASIC1a has thus emerged as drug target with great potential, but no drugs are currently available that specifically target the channels under pathological conditions. A few known neuropeptides modulate ASIC1a and could thus serve as scaffolds for a new generation of ASIC1a-selective drugs to, for example, treat pain without the typical downsides of opioids. Advances have, however, been hampered by the limited understanding of detailed protein-peptide interactions. Thus, the aim of the proposed project is to directly characterize the binding of the neuropeptide Big Dynorphin to ASIC1a in real time. Here, I will use a unique in-house developed high-sensitivity fluorescence patch-clamp electrophysiology setup and establish a protocol for a FRET-based ligand-binding assay. Together with site-directed mutagenesis, this approach will be able to identify state-dependent binding sites and key interactions, and allow direct analysis of binding affinity and kinetics under pathological conditions; all in intact membranes and with unprecedented (microsecond) temporal resolution. This information will aid future design of ASIC inhibitors with the potential to treat chronic pain and ischemia. The technology developed for this work will also enable ligand-binding studies of other membrane proteins in living cells and with high temporal resolution and will thus be of great potential value for a broad field. The project will expand my existing electrophysiology skills and add highly versatile expertise in fluorescent measurements. I thus anticipate my project to have significant personal and scientific impact beyond the scope of this proposal.

Оригинален текст от CORDIS (на английски).

Участници

KOBENHAVNS UNIVERSITET · KOBENHAVNКоординаторДания