H2020Индивидуална стипендия2022–2024

Illuminate-AMR · Single-molecule visualization of individual multi-drug efflux pump activity in living bacteria with nanophotonic biochips

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

Период: 2022-01-01 → 2024-05-22
Финансиране от ЕС: 174 806 €
Участници: 1
Схема: MSCA-IF

Линиите свързват координатора с партньорите.

Накратко на български

Нанофотонни биочипове ще се използват за наблюдение на това как отделни бактерии (като E. coli) изпомпват антибиотици от клетките си. Това помага за разбирането на механизмите на лекарствената резистентност, за да се разработят по-ефективни стратегии за лечение на инфекции.

Този кратък обзор е генериран от изкуствен интелект

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

Резултати накратко

Single-molecule visualization of individual multi-drug efflux pump activity in living bacteria with nanophotonic biochips

The problem being addressed in this project is multidrug resistance (MDR) in bacteria, which poses a significant threat to global health. MDR arises when bacteria develop the ability to resist multiple antimicrobial drugs, reducing the effectiveness of treatments and complicating the management of infections. A key mechanism behind this resistance is drug efflux, where bacteria actively pump out antibiotic molecules, decreasing their intracellular concentration and rendering treatments ineffective. Nevertheless, current imaging techniques are insufficient to visualize and understand the molecular processes driving this resistance, particularly at the bacterial membrane level. This technological gap similarly limits research in other membrane-controlled bioprocesses such as viral infections, cell signaling, and nutrient acquisition. Thus, a deeper understanding of the mechanisms behind drug resistance is essential for maintaining the efficacy of existing drugs and developing novel therapeutic strategies to combat resistant infections. To meet this challenge, the project objectives are set to development and testing of a photonic biochip-based imaging platform for visualization and tracking of membrane-bound proteins i.e. multidrug efflux pumps in individual Escherichia coli cells. The biochip is designed for use with conventional light microscopes, making the single-molecule imaging accessible to many labs without the need to invest in costly and difficult-to-handle equipment. The project further aims to establish a direct assay for imaging and analyzing the activity of multidrug efflux pumps in E. coli cell membrane at the single-cell level. In addition, integration of the nanophotonic chip platform with a microfluidic control system is envisioned. Lastly, performance of the proposed imaging platform in detailing the resistance mechanisms in genetically engineered E. coli cells is to be compared with current single-molecule imaging methods on an industrial scale. To this end, the applications of this platform can be expanded in life sciences and material research, with potential integration into medical diagnostics and other industrial sectors.

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

Цел на проекта

Nanophotonic chips hold great promise in imaging by providing molecular-level insights into biological processes occurring in live cells. By tuning their design, it becomes possible to selectively modify light at their surface such that they can be used to study the properties of biomolecules and cells at high resolution near the interface. On-chip decoupling of the excitation and emission light at multiple wavelengths is however a challenge in optical microscopy. The aim of this project is to substantially improve imaging of membrane processes in cells with biochips and then study population dynamics of multi-drug efflux pump activity in antimicrobial resistant bacteria by widefield nanoscopy. The objectives are (i) to develop and test a new nanophotonic biochip with which to visualize and track membrane bound proteins and their activity in single living bacterial cells under common light microscopes; (ii) to develop a direct assay for imaging the presence and activity of multi-drug efflux pumps in bacterial membranes at the single-cell level; and (iii) to implement and compare the biochip platform with modern single-molecule approaches. This imaging platform is based on encoding several precise periodicities to the surface of the biochip in order to diffraction-couple the light to traveling surface waves at multiple resonant wavelengths. This biochip will be a superior alternative to expensive TIRF microscopes that have a limited field of view, and will practically offer an improved evenness of illumination, smaller footprint, and a more accurate knowledge of penetration depth of the evanescent field. The biochip will be used to elucidate how phenotypic heterogeneity in multi-drug efflux pump activity contributes to bacterial survival and thus help improve current antibiotic treatment schemes. Such technology will result in immediate benefits for integrative cellular and molecular biology research communities, and thus has a strong potential for commercialization.

Оригинален текст от CORDIS (на английски).

Участници

BUNDESANSTALT FUER MATERIALFORSCHUNG UND -PRUEFUNG · BerlinКоординаторГермания