National Research Council of Italy

Institute of Biosciences and BioResources

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Gabriella Pocsfalvi

Role: Research Director
Section: Researchers and Technologists
Division: Naples
Tel: (39) 081-6132570-081-6132585

Extracellular Vesicles and Mass Spetrometry Group

Research interest is focused on i) extracellular vesicles (EVs) and ii.) the development of mass spectrometry-based methods for structural biology and the application of proteomics to protein secretion. Our approach relies on the integration of a range of disciplines (mass spectrometry, chemistry, biochemistry and bioinformatics) to isolate EVs and to increase the overall power of molecular characterization of their biocargo. Pocsfalvi is leader of the Extracellular Vesicles and Mass Spetrometry (EVs-MS) group at IBBR (Napoli). EVsMS group houses hypheneted 2D-nano-HPLC-ESI-QTOF and GC-QqQ mass spectrometers, a protein sequencer and an array of advanced separation techniques. We are actively involved in numerous collaborations, ongoing European projects (see below) and we provide internal service supports in

1. mass spectrometry and proteomics

2. protein and peptide sequencing


Activities of EVs-MS group are focused on the following topics:

Protein secretion and extracellular vesicles

We have a long term interest in the application of proteomics to the characterization of archaeal, bacterial and eukaryotic cell secretomes and secretion mechanisms. Proteins that secreted from cells to the extracellular microenvironment are involved in many biological processes, such as cell to cell communication, cell adhesion, cell migration, invasion and virulence. In this context, we have characterized virulence factors including bacterial protein toxins secreted by pathogenic strains of S. aureus. More recently, we gained knowledge on transport mechanisms in archaea using a combined proteomics and bioinformatics approach.

Cellular secretion of small, membrane enclosed extracellular vesicles (EVs) is a relatively new field but due to their role in intercellular communication is of great interest. We use our in-house developed method for the purification of and quantitative proteome analysis of EVs. We isolate nano-sized vesicles from various sources such as cell cultures, biofluids and plants. Urinary EVs research in our laboratory focuses on different renal diseases (kidney cancer, diabetes type 2 and polycystic kidney disease) using labeling-based quantitative proteomics to identify potential biomarkers that reflect the physiological and/or pathological status of the kidney.

Plants are also secret EVs-like vesicles. Current work in our laboratory targets EVs-like vesicles isolated from different fruit juices.

Affinity mass spectrometry

The EVsMS group is interested in the development of new strategies for targeted molecular analysis. In particular we set-up a solid-phase magnetic immunocapture-based mass spectrometry method for the detection of bacterial protein toxins. In addition, a single-tube batchwise method employing nano-sized TiO2 and ZrO2 particles was developed to enrich phosphopeptides and characterize protein phosphorylation status.

Natural vesicles-based nano-sized delivery system

The EVsMS group is actively researching the biotechnological and biomedical exploitation of nanovesicles derived from natural resources by transfering unique know-how of the group acquired in the isolation and characterization of EVs. Novel isolation techniques and fabriacation of vesicles loaded with selected drugs, nutraceuticals and cosmeceuticals are beeing the center of the attention of two recently founded European projects.

Ongoing Projects:

EV-C@p - Progetti di Grande Rilevanza supported in part by the Italian Ministry of Foreign Affairs and International Cooperation”, grant number KR23GR03. "Plant-derived Extracellular Vesicles as novel biotechnological platform for biopharma applications” (2023-2026)

Going beyond the state-of-the-art and relying on highly interconnected approaches mastered by the applicants, in EV-C@p will setup a novel vegetable-based system for the production of nanovesicles and exploit them as a biotechnological platform for biopharma applications. This will be achieved through the isolation of EVs from organically grown tomato and panax ginseng and by the establishment of their in vitro cell cultures to biomanifacture EVs.

By applying emerging biotechnology we will develop engineered EV-based solutions to create next generation nano-delivery systems. The native and the engineered plant EVs will be exploited for their health promoting anticancer and anti-inflammatory activities in vitro. To achieve the above objectives the EV-C@p consortium was created with the participation of two research groups (KHU and CNR) and two SMEs partners, ImmunoVeg from Italy and NIOBiopharmaceuticals from Korea. The project will strengthen the already existing collaborations between the two PIs and create new ones with the SMEs to provide a solid cornerstone for further research efforts of the participants and to make a prompt translation of the research results into new technology and products after the ending of the project. An ambitious staff exchange programme for the transfer of scientific and transferable skills, dissemination and outreach activities were implemented into the scientific plan.

VES4US - Horizon 2020 FETOPEN project "Extracellular vesicles from a natural source for tailormade nanomaterials" (2018-2021)

The VES4US project is an European initiative funded by the Horizon2020 Program of the European Commission under Future Emerging Technologies (FET) call. Its main objective aims at generating a broad range of radically new high-value products in the fields of nutrition, cosmetics and health sciences based on natural source-derived extracellular vesicles (EVs), which could be used as new generation vehicles for specific molecular delivery.

The EVsMS group leads WP2 of VES4US project that will produce of natural source derived vesicles with radically new technology, as well as Task 3.3. "Characterisation of protein and small molecular biocargo of vesicles”.

nanoTOM - Horizon 2020 MSCA-IF "Conversion of natural plant nanovesicles into nutraceutical delivery system" (2018-2020)

Novel encapsulation approaches to create alternative delivery options for nutraceuticals are emerging as a promising strategy to enhance the bioavailability of poorly absorbed active food ingredients. In this context, nanoTOM aims to exploit edible plant derived nanovesicles and use them as vehicles for the encapsulation, protection, release and bioavailability enhancement of selected nutraceuticals. Plant cells secret phospholipid membrane-surrounded vesicles morphologically similar to mammalian extracellular vesicles. Exploitation of plant nanovesicles is promising although hampered by i) their difficult isolation and ii) the lack of knowledge of their biogenesis, molecular architecture, uptake and biological effects. The experienced researcher is a chemist with strong academic and pharmaceutical background in the isolation and analysis of
bioactive compounds from medicinal plants who team-up with the Institute of Biosciences and BioResources (IBBR) with considerable expertise in extracellular vesicle research to realize a uniquely interdisciplinary research program. The research proposed here will realize the following concrete objectives: 1. Set-up an integrated analytical pipeline for the isolation, characterization, encapsulation, uptake and toxicological profiling of plant nanovesicles. 2. Use the pipeline to exploit different Solanum lycopersicum (tomato) nanovesicle populations regarding secretion mechanism, heterogeneity, biocargo composition, nutraceutical and encapsulation properties.

Nutri-C@rgo: Characterization of plant secreted nanovesicles (2016-2018)

A joint research project of the CNR ant he Hungarian Academy of Sceinces.

to study the molecular cargo of EVs isolated from fruit juices of the citrus family. The molecular data is produced and used to define the molecular cargo of citrus derived EVs, their main differences to those derived from mammalian cells. The study focuses predominantly on the protein cargo, but we will explore other components, like lipids and possibly glycosylated products as well. The molecular data will be used to answer the following questions: i.) what is the molecular cargo of citrus derived EVs? ii) what are the differences among EVs isolated from different citrus species? iii.) how much does the protein and lipid cargo of plant EVs differ from that of mammalian cells? and, iv.) what are the possible interlocutors involved in the crosstalk between plant EVs and mammalian cells? The obtained data sets will be deposited into two manually curated web-based databases, EVpedia and Vesiclepedia, that are publicly available and at the moment lack such data.


Selected Publications

(full list available at CNR People)

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