Adipose Mesenchymal Extracellular Vesicles as Alpha-1-Antitrypsin Physiological Delivery Systems for Lung Regeneration
Elia Bari,
Ilaria Ferrarotti,
Dario Di Silvestre,
Pietro Grisoli,
Valentina Barzon,
Alice Balderacchi,
Maria Luisa Torre,
Rossana Rossi,
Pierluigi Mauri,
Angelo Guido Corsico,
Sara Perteghella
Affiliations
Elia Bari
Department of Drug Sciences, University of Pavia, Viale Taramelli 12, 27100 Pavia, Italy
Ilaria Ferrarotti
Center for Diagnosis of Inherited Alpha1-antitrypsin Deficiency, Department of Internal Medicine and Therapeutics, Pneumology Unit, IRCCS San Matteo Hospital Foundation, University of Pavia, 27100 Pavia, Italy
Dario Di Silvestre
Institute for Biomedical Technologies, F.lli Cervi 93, 20090 Segrate, Milan, Italy
Pietro Grisoli
Department of Drug Sciences, University of Pavia, Viale Taramelli 12, 27100 Pavia, Italy
Valentina Barzon
Center for Diagnosis of Inherited Alpha1-antitrypsin Deficiency, Department of Internal Medicine and Therapeutics, Pneumology Unit, IRCCS San Matteo Hospital Foundation, University of Pavia, 27100 Pavia, Italy
Alice Balderacchi
Center for Diagnosis of Inherited Alpha1-antitrypsin Deficiency, Department of Internal Medicine and Therapeutics, Pneumology Unit, IRCCS San Matteo Hospital Foundation, University of Pavia, 27100 Pavia, Italy
Maria Luisa Torre
Department of Drug Sciences, University of Pavia, Viale Taramelli 12, 27100 Pavia, Italy
Rossana Rossi
Institute for Biomedical Technologies, F.lli Cervi 93, 20090 Segrate, Milan, Italy
Pierluigi Mauri
Institute for Biomedical Technologies, F.lli Cervi 93, 20090 Segrate, Milan, Italy
Angelo Guido Corsico
Center for Diagnosis of Inherited Alpha1-antitrypsin Deficiency, Department of Internal Medicine and Therapeutics, Pneumology Unit, IRCCS San Matteo Hospital Foundation, University of Pavia, 27100 Pavia, Italy
Sara Perteghella
Department of Drug Sciences, University of Pavia, Viale Taramelli 12, 27100 Pavia, Italy
Accumulating evidence shows that Mesenchymal Stem/Stromal Cells (MSCs) exert their therapeutic effects by the release of secretome, made of both soluble proteins and nano/microstructured extracellular vesicles (EVs). In this work, for the first time, we proved by a proteomic investigation that adipose-derived (AD)-MSC-secretome contains alpha-1-antitrypsin (AAT), the main elastase inhibitor in the lung, 72 other proteins involved in protease/antiprotease balance, and 46 proteins involved in the response to bacteria. By secretome fractionation, we proved that AAT is present both in the soluble fraction of secretome and aggregated and/or adsorbed on the surface of EVs, that can act as natural carriers promoting AAT in vivo stability and activity. To modulate secretome composition, AD-MSCs were cultured in different stimulating conditions, such as serum starvation or chemicals (IL-1β and/or dexamethasone) and the expression of the gene encoding for AAT was increased. By testing in vitro the anti-elastase activity of MSC-secretome, a dose-dependent effect was observed; chemical stimulation of AD-MSCs did not increase their secretome anti-elastase activity. Finally, MSC-secretome showed anti-bacterial activity on Gram-negative bacteria, especially for Klebsiella pneumoniae. These preliminary results, in addition to the already demonstrated immunomodulation, pave the way for the use of MSC-secretome in the treatment of AAT-deficiency lung diseases.
