H2020Индивидуална стипендия2017–2019

EV-LNC · Extracellular vesicle-mediated delivery of long non-coding RNA: Implications for vascular repair and regeneration

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

Период: 2017-08-01 → 2019-07-31
Финансиране от ЕС: 195 455 €
Участници: 1
Схема: MSCA-IF-EF-ST

Линиите свързват координатора с партньорите.

Накратко на български

Връзката между гладките мускулни клетки и ендотелните клетки в кръвоносните съдове се анализира чрез преноса на малки молекули (екстрацелулярни везикули) и специфична RNA. Това помага да се разбере как протичат процесите на възстановяване и регенерация при съдови увреждания и белодробна хипертония.

Този кратък обзор е генериран от изкуствен интелект

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

Резултати накратко

Extracellular vesicle-mediated delivery of long non-coding RNA: Implications for vascular repair and regeneration

Objectives: The main aim of this project is to study extracellular vesicle mediated cell-to-cell communication between human smooth muscle cells and endothelial cells, evaluate its relevance in vascular injury in an in vitro model of pulmonary arterial hypertension, and determine the significance of long non-coding RNA in this crosstalk. Methodology: For imaging EV transfer among vascular cells, we will use a pioneer approach based on Cre-loxP recombination which results in a fluorescent colour switch of cells upon EV uptake. Cells will then be sorted according to fluorescence by flow cytometry and analysed by cutting-edge proteomics and bioinformatics, in order to elucidate intercellular signalling triggered by EV transfer. The lncRNAs present in EVs will be analysed by RNA-Seq. Mechanistic insight of enriched lncRNAs in EVs will be evaluated using gain- and loss-of function approaches in vascular cells using lentiviral vectors and GapmeRs/siRNAs, respectively. Integration of all these analyses will provide key information to define implications of EV-mediated delivery of lncRNA for vascular repair and regeneration. Conclusions • PASMCs communicate with PAECs via EVs and the latter take up and translate RNA cargo to synthesize proteins. • PASMC-to-PAECs EV-mediated communication is enhanced by TGF-β1 and IL1-β possibly due to increased uptake of PASMC-EVs activated PAECS. Since these cytokines are increased in pulmonary hypertension, this communication may be enhanced during PAH. • PASMC-EVs are enriched in the transcription factor Zeb1, related to EndoMT, and two TGF-β ligands: GDF11 and TGF-β3, at the RNA level. Zeb1 at the protein level too. • TGF-β1 treated PASMC-EVs increase their levels of paladin and bHLHE40 protein-coding RNAs, involved in EndoMT. • PAECs that take up PASMC-EVs increase their levels of smooth-muscle actin, the most widely used EndoMT maker. Thus, PASMC-EVs can induce EndoMT on PAECs. • PASMC-EVs are enriched in the lncRNA FLJ16779, which is very specific of PASMC-EVs. • TGF-β1 treated PASMC-EVs overexpress mysolid lncRNA. Overexpressed mysolid is specifically sorted into PASMC-EVs and might be transported into PAECs.

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

Цел на проекта

Background: The concept of extracellular vesicles (EVs) has drastically changed from the initial non-functional debris to the current of key mediators of paracrine signalling. The cargoes of EVs comprise all kinds of macromolecules, and recent evidence has demonstrated the presence of long non-coding RNAs (lncRNAs) in such vesicles. These RNA molecules have numerous potential regulatory functions and results obtained so far guide us to presume a determinant role in vascular cell differentiation, proliferation and repair. Besides, the increasing data emerging in the field are significantly changing the way in which we interpret molecular mechanisms driving cardiovascular diseases and offers a brand new set of molecular targets for therapy. For all these reasons, study of lncRNAs in vascular biology and disease is state-of-the-art. Objectives: The main aim of this project is to study extracellular vesicle mediated cell-to-cell communication between human smooth muscle cells and endothelial cells, evaluate its relevance in vascular injury in an in vitro model of pulmonary arterial hypertension, and determine the significance of long non-coding RNA in this crosstalk. Methodology: For imaging EV transfer among vascular cells, we will use a pioneer approach based on Cre-loxP recombination which results in a fluorescent colour switch of cells upon EV uptake. Cells will then be sorted according to fluorescence by flow cytometry and analysed by cutting-edge proteomics and bioinformatics, in order to elucidate intercellular signalling triggered by EV transfer. The lncRNAs present in EVs will be analysed by RNA-Seq. Mechanistic insight of enriched lncRNAs in EVs will be evaluated using gain- and loss-of function approaches in vascular cells using lentiviral vectors and GapmeRs/siRNAs, respectively. Integration of all these analyses will provide key information to define implications of EV-mediated delivery of lncRNA for vascular repair and regeneration.

Оригинален текст от CORDIS (на английски).

Участници

THE UNIVERSITY OF EDINBURGH · EdinburghКоординаторОбединеното кралство