H2020Индивидуална стипендия2021–2023

REMIND EBOV · Role of the mucin-like domain of the Ebola virus in modulating virus-glycosaminoglycan interactions

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

Период: 2021-08-01 → 2023-08-16
Финансиране от ЕС: 191 852 €
Участници: 1
Схема: MSCA-IF

Линиите свързват координатора с партньорите.

Накратко на български

Ролята на муцинподобния домейн на вируса Ебола се анализира чрез проследяване на отделни вирусни частици, за да се види как те се закрепят за клетката. Това помага за по-доброто разбиране на вирологията и оптимизирането на ваксини, базирани на псевдотипове.

Този кратък обзор е генериран от изкуствен интелект

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

Резултати накратко

Role of the mucin-like domain of the Ebola virus in modulating virus-glycosaminoglycan interactions

Single particle tracking (SPT) which allows for the visualization of key steps of the virus life cycle on a single virus particle level, promises to expand the knowledge in the field of virology drastically: it offers the possibility to reveal transient and dynamic processes that are otherwise masked in static or ensemble-averaged measurements. In the context of applying SPT to studying virus attachment and entry into host cells, virus pseudotypes, i.e., particles displaying the structural core of one virus and the functional envelope glycoprotein (GP) of a heterologous virus of interest, are promising candidates: they are compatible with biosafety level -2 conditions and can be easily labelled. However, heterogeneities in the GP distributions represent a significant hurdle for single particle applications, as they may affect the particle behaviour. Accordingly, it is important to characterize pseudotypes on an individual particle level, to optimize production, labelling and data acquisition strategies. In this project, fluorescent pseudotypes of the deadly filovirus Ebola were produced, using a lentiviral pseudotyping system with a mCherry-tagged viral core. A workflow for their in-depth characterization on a single particle level is proposed. Additionally, the particles were used in single particle kinetic assays, to test the hypothesis that mucin-like domain of Ebola, a highly glycosylated region of the protein, is important in modulating the attachment and detachment of the virus from the cell surface. A thorough characterization of pseudotype properties and the production of more homogenous samples will strengthen the interpretations of experimental results within virology and also contribute to the optimization of pseudotype-based vaccines. In addition, understanding the mechanisms by which virus interactions at the cell surface are modulated, can help developing therapeutics for future outbreaks.

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

Цел на проекта

Ebola virus, one of the deadliest human pathogens, is a known candidate for severe outbreaks and has caused several thousand deaths in the more recent outbreaks alone. To fight against it, a detailed knowledge about its viral life cycle is fundamental to the development of efficient vaccines and drugs. In this project I suggest to investigate the role of the mucin-like domain (MLD) of the viral glycoprotein (GP) in modulating virus attachment, detachment and diffusion on glycosaminoglycans (GAGs), responsible for recruiting the virus at the cell surface. To do so, I will generate GP-containing pseudotyped viruses, mimicking the tropism of the pathogen. Specifically, I will compare the GP of the Zaire strain of EBOV, an MLD-deleted mutant and a natural mutant that occurred during the West Africa outbreak (2013-2016) that is reported to have an increased tropism for human cells. I will use advanced biophysical techniques to examine the interactions on a molecular level as well as on the cellular level. On a molecular level, I will study the binding strength of individual bonds formed between the GP and GAGs using force spectroscopy. In addition, I will investigate the attachment and detachment of virus particles from GAGs immobilized on a glass surface in a biomimetic fashion, using total internal fluorescence microscopy. Proceeding to a more physiological model using living cells, I plan to study the diffusion behavior at the cell surface of pseudotyped viruses carrying the various mutations in their MLD. Stepping up in complexity, in the last part of the project, I will investigate the role of the MLD in modulating the ability of the virus to cross the glycocalyx, the sugar coat of cells, by employing 3D tracking. Taken together this project will lead to a better understanding on how viral particle migrate on the cell surface and how the interactions function on a molecular level.

Оригинален текст от CORDIS (на английски).

Участници

UMEA UNIVERSITET · UMEAКоординаторШвеция