Isolation, characterization and potential role in beta cell-endothelium cross-talk of extracellular vesicles released from human pancreatic islets

Federico Figliolini, Vincenzo Cantaluppi, Michela De Lena, Silvia Beltramo, Renato Romagnoli, Mauro Salizzoni, Raffaella Melzi, Rita Nano, Lorenzo Piemonti, Ciro Tetta, Luigi Biancone, Giovanni Camussi

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Abstract

The cross-talk between beta cells and endothelium plays a key role in islet physiopathology and in the revascularization process after islet transplantation. However, the molecular mechanisms involved in this cross-talk are not fully elucidated. Extracellular vesicles (EVs) are secreted membrane nanoparticles involved in inter-cellular communication through the transfer of proteins and nucleic acids. The aims of this study were: 1) isolation and characterization of EVs from human islets; 2) evaluation of the pro-angiogenic effect of islet-derived EVs on human islet endothelial cells (IECs). EVs were isolated by ultracentrifugation from conditioned medium of human islets and characterized by nanotrack analysis (Nanosight), FACS, western blot, bioanalyzer, mRNA/microRNA RT-PCR array. On IECs, we evaluated EV-induced insulin mRNA transfer, proliferation, resistance to apoptosis, in vitro angiogenesis, migration, gene and protein profiling. EVs sized 236±54 nm, expressed different surface molecules and islet-specific proteins (insulin, C-peptide, GLP1R) and carried several mRNAs (VEGFa, eNOS) and microRNAs (miR-27b, miR-126, miR-130 and miR-296) involved in beta cell function, insulin secretion and angiogenesis. Purified EVs were internalized into IECs inducing insulin mRNA expression, protection from apoptosis and enhancement of angiogenesis. Human islets release biologically active EVs able to shuttle specific mRNAs and microRNAs (miRNAs) into target endothelial cells. These results suggest a putative role for islet-derived EVs in beta cell-endothelium cross-talk and in the neoangiogenesis process which is critical for engraftment of transplanted islets.

Original languageEnglish
Article numbere102521
JournalPLoS One
Volume9
Issue number7
DOIs
Publication statusPublished - Jul 16 2014

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islets of Langerhans
angiogenesis
endothelium
Islets of Langerhans
endothelial cells
Endothelium
Endothelial cells
microRNA
Messenger RNA
MicroRNAs
insulin
Insulin
apoptosis
cells
Endothelial Cells
cell communication
ultracentrifugation
proteins
insulin secretion
Apoptosis

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Medicine(all)

Cite this

Figliolini, F., Cantaluppi, V., De Lena, M., Beltramo, S., Romagnoli, R., Salizzoni, M., ... Camussi, G. (2014). Isolation, characterization and potential role in beta cell-endothelium cross-talk of extracellular vesicles released from human pancreatic islets. PLoS One, 9(7), [e102521]. https://doi.org/10.1371/journal.pone.0102521

Isolation, characterization and potential role in beta cell-endothelium cross-talk of extracellular vesicles released from human pancreatic islets. / Figliolini, Federico; Cantaluppi, Vincenzo; De Lena, Michela; Beltramo, Silvia; Romagnoli, Renato; Salizzoni, Mauro; Melzi, Raffaella; Nano, Rita; Piemonti, Lorenzo; Tetta, Ciro; Biancone, Luigi; Camussi, Giovanni.

In: PLoS One, Vol. 9, No. 7, e102521, 16.07.2014.

Research output: Contribution to journalArticle

Figliolini, F, Cantaluppi, V, De Lena, M, Beltramo, S, Romagnoli, R, Salizzoni, M, Melzi, R, Nano, R, Piemonti, L, Tetta, C, Biancone, L & Camussi, G 2014, 'Isolation, characterization and potential role in beta cell-endothelium cross-talk of extracellular vesicles released from human pancreatic islets', PLoS One, vol. 9, no. 7, e102521. https://doi.org/10.1371/journal.pone.0102521
Figliolini, Federico ; Cantaluppi, Vincenzo ; De Lena, Michela ; Beltramo, Silvia ; Romagnoli, Renato ; Salizzoni, Mauro ; Melzi, Raffaella ; Nano, Rita ; Piemonti, Lorenzo ; Tetta, Ciro ; Biancone, Luigi ; Camussi, Giovanni. / Isolation, characterization and potential role in beta cell-endothelium cross-talk of extracellular vesicles released from human pancreatic islets. In: PLoS One. 2014 ; Vol. 9, No. 7.
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