Gabriella Pocsfalvi

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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

3. GC-MS/MS

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)

Protein Biocargo and Anti-Inflammatory Effect of Tomato Fruit-Derived Nanovesicles Separated by Density Gradient Ultracentrifugation and Loaded with Curcumin

Mammadova R, Maggio S, Fiume I, Bokka R, Moubarak M, Gellén G, Schlosser G, Adamo G, Bongiovanni A, Trepiccione F, Guescini M, Pocsfalvi G

Year: 2023

Mysterious travellers: Identification of viral and host defense-related components in tomato fruit-derived nanovesicles

Sabetta W, Mammadova R, Kralj-iglic V, Schlosper G, Pocsfalvi G, Cillo F

Year: 2022

Nanoalgosomes: Introducing extracellular vesicles produced by microalgae

Adamo G, Fierli D, Romancino DP, Picciotto S, Barone ME, Aranyos A, Božič D, Morsbach S, Raccosta S, Stanly C, Paganini C, Gai M, Cusimano A, Martorana V, Noto R, Carrotta R, Librizzi F, Randazzo L, Parkes R, Capasso Palmiero U, Rao E, Paterna A, Santonicola P, Iglič A, Corcuera L, Kisslinger A, Schiavi ED, Liguori GL, Landfester K, Kralj-Iglič V, Arosio P, Pocsfalvi G, Touzet N, Manno M, Bongiovanni A

Year: 2021

Isolation of extracellular vesicles from microalgae: towards the production of sustainable and natural nanocarriers of bioactive compounds

Picciotto S, Barone ME, Fierli D, Aranyos A, Adamo G, Božič D, Romancino DP, Stanly C, Parkes R, Morsbach S, Raccosta S, Paganini C, Cusimano A, Martorana V, Noto R, Carrotta R, Librizzi F, Capasso Palmiero U, Santonicola P, Iglič A, Gai M, Corcuera L, Kisslinger A, Di Schiavi E, Landfester K, Liguori GL, Kralj-Iglič V, Arosio P, Pocsfalvi G, Manno M, Touzet N, Bongiovanni A

Year: 2021

Bilateral project CNR-CINVESTAV mEVChip: A microfluidic device to assess the anti-inflammatory effects of ginger-derived nanovesicles on mammalian cells

Pocsfalvi G, Fiume I, Ramos AP, J-L G-C, Amador Hernandez J, Rodriguez R

Year: 2021

Pursuing mechanisms of extracellular vesicle formation. Effects of sample processing

Božič D, Hočevar M, Kononenko V, Jeran M, Štibler U, Fiume I, Pajnič M, PaÄàen L, Kogej K, Drobne D, Iglič A, Pocsfalvi G, Kralj-Iglič V

Year: 2020

Plant roots release small extracellular vesicles with antifungal activity

De Palma M, Ambrosone A, Leone A, Del Gaudio P, Ruocco M, Turiák L, Bokka R, Fiume I, Tucci M, Pocsfalvi G

Year: 2020

The human EV membranome

Ramos Juarez AP, Trepiccione F, Capasso G, Pocsfalvi G

Year: 2020

Potential allergenicity of Medicago sativa investigated by a combined IgE-binding inhibition, proteomics and in silico approach

Yakhlef M, Giangrieco I, Ciardiello MA, Fiume I, Mari A, Souiki L, Pocsfalvi G

Year: 2020

Bacterial IAA-delivery into Medicago root nodules triggers a balanced stimulation of C and N metabolism leading to biomass increase

Defez R, Andreozzi A, Romano S, Pocsfalvi G, Fiume I, Esposito R, Angelini C, Bianco C

Year: 2019

Bacterial IAA-Delivery into Medicago Root Nodules Triggers a Balanced Stimulation of C and N Metabolism Leading to a Biomass Increase

Defez R, Andreozzi A, Romano S, Pocsfalvi G, Fiume I, Esposito R, Angelini C, Bianco C

Year: 2019

Scalable Production and Isolation of Extracellular Vesicles: Available Sources and Lessons from Current Industrial Bioprocesses

Paganini Carolina, Palmiero Umberto Capasso, Pocsfalvi Gabriella, Touzet Nicolas, Bongiovanni Antonella, Arosio Paolo

Year: 2019

Dissection of protein cargo of Citrus fruit juice sac cells-derived vesicles reveals heterogeneous transport and extracellular vesicles subpopulations

Pocsfalvi G, Turiák L, Ambrosone A, Del Gaudio P, Puska G, Fiume I, Silvestre T, Vékey K

Year: 2018

Best practice of identification and proteomic analysis of extracellular vesicles in human health and disease

Sódar BW, Kovács Á, Visnovitz T, Pállinger É, Vékey K, Pocsfalvi G, Turiák L, Buzás EI

Year: 2017

Introducing transgalactosylation activity into a family 42 β-galactosidase

Strazzulli A, Cobucci-Ponzano B, Carillo S, Bedini E, Corsaro MM, Pocsfalvi G, Withers SG, Rossi M, Moracci M

Year: 2017

Evidence-based clinical use of nanoscale extracellular vesicles in nanomedicine

Fais S, O’Driscoll L, Borras FE, Buzas E, Camussi G, Cappello F, Carvalho J, Cordeiro da Silva A, Del Portillo H, El Andaloussi S, Ficko Trček T, Furlan R, Hendrix A, Gursel I, Kralj-Iglic V, Kaeffer B, Kosanovic M, Lekka ME, Lipps G, Logozzi M, Marcilla A, Sammar M, Llorente A, Nazarenko I, Oliveira C, Pocsfalvi G, Rajendran L, Raposo G, Rohde E, Siljander P, van Niel G, Vasconcelos MH, Yáñez-Mó M, Yliperttula ML, Zarovni N, Zavec AB, Giebel B

Year: 2016

Chromatography and its hyphenation to mass spectrometry for extracellular vesicle analysis

Pocsfalvi G, Stanly C, Fiume I, Vékey K

Year: 2016

Mass spectrometry of extracellular vesicles

Pocsfalvi G, Stanly C, Vilasi A, Fiume I, Capasso G, Turiák L, Buzas EI, Vékey K

Year: 2016

Isolation and characterisation of a novel alpha-amylase from the extreme haloarchaeon Haloterrigena turkmenica

Santorelli M, Maurelli L, Pocsfalvi G, Fiume I, Squillaci G, La Cara F, Del Monaco G, Morana A

Year: 2016

Isolation of exosome-like vesicles from plants by ultracentrifugation on sucrose/deuterium oxide (D₂O) density cushions

Stanly C, Fiume I, Capasso G, Pocsfalvi G

Year: 2016

Applying extracellular vesicles based therapeutics in clinical trials - an ISEV position paper

Lener T, Gimona M, Aigner L, Börger V, Buzas E, Camussi G, Chaput N, Chatterjee D, Court FA, del Portillo HA, O’driscoll L, Fais S, Falcon-Perez JM, Felderhoff-Mueser U, Fraile L, Song Gho Y, Görgens A, Gupta RC, Hendrix A, Hermann DM, Hill AF, Hochberg F, Horn PA, de Kleijn D, Kordelas L, Kramer BW, Krämer-Albers EM, Laner-Plamberger S, Laitinen S, Leonardi T, Lorenowicz MJ, Kiang Lim S, Lötvall J, Maguire CA, Marcilla A, Nazarenko I, Ochiya T, Patel T, Pedersen S, Pocsfalvi G, Pluchino S, Quesenberry P, Reischl IG, Rivera FJ, Sanzenbacher R, Schallmoser K, Slaper-Cortenbach I, Strunk D, Tonn T, Vader P, van Balkom BW, Wauben M, El Andaloussi S, Théry C, Rohde E, Giebel B

Year: 2015

Urinary extracellular vesicles as reservoirs of altered proteins during the pathogenesis of polycystic kidney disease

Pocsfalvi G, Raj DA, Fiume I, Vilasi A, Trepiccione F, Capasso G

Year: 2015

EVpedia: a community web portal for extracellular vesicles research

Kim DK, Lee J, Kim SR, Choi DS, Yoon YJ, Kim JH, Go G, Nhung D, Hong K, Jang SC, Kim SH, Park KS, Kim OY, Park HT, Seo JH, Aikawa E, Baj-Krzyworzeka M, van Balkom BW, Belting M, Blanc L, Bond V, Bongiovanni A, Borras FE, Buee L, Buzas EI, Cheng L, Clayton A, Cocucci E, Dela Cruz CS, Desiderio DM, Di Vizio D, Ekstrom K, Falcon-Perez JM, Gardiner C, Giebel B, Greening DW, Gross JC, Gupta D, Hendrix A, Hill AF, Hill MM, Nolte-’t Hoen E, Hwang DW, Inal J, Jagannadham MV, Jayachandran M, Jee YK, Jorgensen M, Kim KP, Kim YK, Kislinger T, Lasser C, Lee DS, Lee H, van Leeuwen J, Lener T, Liu ML, Lotvall J, Marcilla A, Mathivanan S, Moller A, Morhayim J, Mullier F, Nazarenko I, Nieuwland R, Nunes DN, Pang K, Park J, Patel T, Pocsfalvi G, Del Portillo H, Putz U, Ramirez MI, Rodrigues ML, Roh TY, Royo F, Sahoo S, Schiffelers R, Sharma S, Siljander P, Simpson RJ, Soekmadji C, Stahl P, Stensballe A, Stepien E, Tahara H, Trummer A, Valadi H, Vella LJ, Wai SN, Witwer K, Yanez-Mo M, Youn H, Zeidler R, Gho YS

Year: 2014

Human Hsp60 with Its Mitochondrial Import Signal Occurs in Solution as Heptamers and Tetradecamers Remarkably Stable over a Wide Range of Concentrations

Vilasi S, Carrotta R, Mangione MR, Campanella C, Librizzi F, Randazzo L, Martorana V, Marino Gammazza A, Ortore MG, Vilasi A, Pocsfalvi G, Burgio G, Corona D, Palumbo Piccionello A, Zummo G, Bulone D, Conway de Macario E, Macario AJ, San Biagio PL, Cappello F

Year: 2014

Surface-exposed Glycoproteins of Hyperthermophilic Sulfolobus solfataricus P2 Show a Common N-Glycosylation Profile

Palmieri G, Balestrieri M, Peter-Katalinic J, Pohlentz G, Rossi M, Fiume I, Pocsfalvi G

Year: 2013

Enrichment specificity of micro and nano-sized titanium and zirconium dioxides particles in phosphopeptide mapping

Vilasi A, Fiume I, Pace P, Rossi M, Pocsfalvi G

Year: 2013

Urinary exosomes for protein biomarker research

Amal Raj DA, Fiume I, Capasso G, Pocsfalvi G

Year: 2012

Structural and Functional Insights into Aeropyrum pernix OppA, a Member of a Novel Archaeal OppA Subfamily

Balestrieri M, Gogliettino M, Fiume I, Pocsfalvi G, Catara G, Rossi M, Palmieri G

Year: 2011

Detection of bacterial protein toxins by solid phase magnetic immunocapture and mass spectrometry

Pocsfalvi G, Schlosser G

Year: 2011

A new archaeal beta-glycosidase from Sulfolobus solfataricus: seeding a novel retaining beta-glycan-specific glycoside hydrolase family along with the human non-lysosomal glucosylceramidase GBA2.

Cobucci-Ponzano B, Aurilia V, Riccio G, Henrissat B, Coutinho PM, Strazzulli A, Padula A, Corsaro MM, Pieretti G, Pocsfalvi G, Fiume I, Cannio R, Rossi M, Moracci M

Year: 2010