Extracellular Vesicles from Human Induced Pluripotent Stem Cells Reduce Lung Ischemia-Reperfusion Injury via Cytoprotective Mechanisms in both 2D Cultures and a Human Airway Organoid Model.
11 affiliations
- 1Research Department, IRCCS ISMETT (Istituto Mediterraneo per i Trapianti e Terapie ad alta specializzazione), Palermo, Italy.
- 2Unit of Cell and Gene Therapies, Fondazione IRCCS Ca' Granda Ospedale Maggiore Policlinico, Milan, Italy.
- 3Ri.MED Foundation, Palermo, Italy.
- 4Università degli Studi di Enna "Kore", Enna, Italy.
- 5Departments of Surgery and Bioengineering, McGowan Institute, University of Pittsburgh, Pittsburgh, USA.
- 6MePreCC, Università degli studi di Palermo, Palermo, Italy.
- 7Laboratory of Gynecological Preclinical Oncology, Experimental Oncology Department, Istituto di Ricerche Farmacologiche Mario Negri IRCCS, Milan, Italy.
- 8Dipartimento di Medicina Clinica, Sanità Pubblica, Scienze della Vita e dell'Ambiente, Università de L'Aquila, L'Aquila, Italy.
- 9Unit of Thoracic Surgery and Lung Transplantation, IRCCS ISMETT (Istituto Mediterraneo per i Trapianti e Terapie ad alta specializzazione), Palermo, Italy.
- 10Unit of Thoracic Surgery and Transplantation, Fondazione IRCCS Ca' Granda Ospedale Maggiore Policlinico, Milan, Italy.
- 11Department of Medical-Surgical Physiopathology and Transplantation, University of Milan, Milan, Italy.
Abstract
Lung transplantation (LTx) remains the only curative option for patients with end-stage pulmonary diseases. However, long-term outcomes are compromised by ischemia-reperfusion injury (IRI), a major contributor to primary graft dysfunction and chronic lung allograft impairment. Current pharmacological and technological strategies provide only partial protection, highlighting the need for novel therapeutic approaches. In this study, we investigated the therapeutic potential of extracellular vesicles derived from human induced pluripotent stem cells (hiPSC-EV) in a two-dimensional (2D) in vitro model of pulmonary IRI using A549 lung epithelial cells. To better recapitulate the in vivo airway microenvironment, we also employed a human airway organoid (hAO) model. Proteomic analysis of hiPSC-EV identified 1,698 proteins enriched in pathways relevant to apoptosis regulation, oxidative stress responses, and inflammatory signaling. In the 2D IRI model, the administration of hiPSC-EV during reperfusion significantly improved cell viability, reduced lactate dehydrogenase (LDH) release, attenuated reactive oxygen species (ROS) accumulation, and decreased apoptosis and necrosis. Furthermore, hiPSC-EV downregulated pro-inflammatory mediators (NFkB1, TNFα, IL1β, IL8, MCP1) while upregulating anti-inflammatory and anti-apoptotic key factors (IL1RA, BCL2). In the 3D model, hiPSC-EV administration preserved organoid structure and viability under IRI conditions, attenuated transcriptional and protein-level expression of pro-inflammatory and oxidative stress markers (IL1β, IL8, HMGB1, HMOX1), and enhanced anti-inflammatory (IL10) and cytoprotective (HSP27, HSP70) responses. Immunofluorescence further revealed that hiPSC-EV reduced activation of apoptotic and pyroptotic pathways. Moreover, intravenous administration of hiPSC-EV in mice showed no adverse safety signals, including tumorigenicity. Together, these findings demonstrate that hiPSC-EV exert broad cytoprotective effects in pulmonary IRI through coordinated regulation of inflammation, oxidative stress, and cell death. Their scalability, reproducibility, and favorable safety profile underscore their promise as innovative cell-free therapeutics to improve outcomes in LTx.
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BibTeX
@article{tinnirello2026extracellular,
title = {{Extracellular Vesicles from Human Induced Pluripotent Stem Cells Reduce Lung Ischemia-Reperfusion Injury via Cytoprotective Mechanisms in both 2D Cultures and a Human Airway Organoid Model.}},
author = {Rosaria Tinnirello and Maria Chiara Iachini and Maura Cimino and Noemi Elia and Margot Lo Pinto and Simone Dario Scilabra and Gioacchin Iannolo and Giovanni Zito and Roberta Vazzana and Nicola Cuscino and Matteo Bulati and Tuğba Dursun Usal and Federica Cosentino and Antonio D'amore and Federica Guffanti and Giovanna Damia and Vincenza Dolo and Massimo Pinzani and Pier Giulio Conaldi and Alessandro Bertani and Lorenzo Rosso and Lorenza Lazzari and Vitale Miceli},
journal = {International journal of biological sciences},
year = {2026},
month = oct,
doi = {10.7150/ijbs.134686},
url = {https://europepmc.org/article/MED/42910275},
note = {CC BY},
}
RIS
TY - JOUR
TI - Extracellular Vesicles from Human Induced Pluripotent Stem Cells Reduce Lung Ischemia-Reperfusion Injury via Cytoprotective Mechanisms in both 2D Cultures and a Human Airway Organoid Model.
AU - Rosaria Tinnirello
AU - Maria Chiara Iachini
AU - Maura Cimino
AU - Noemi Elia
AU - Margot Lo Pinto
AU - Simone Dario Scilabra
AU - Gioacchin Iannolo
AU - Giovanni Zito
AU - Roberta Vazzana
AU - Nicola Cuscino
AU - Matteo Bulati
AU - Tuğba Dursun Usal
AU - Federica Cosentino
AU - Antonio D'amore
AU - Federica Guffanti
AU - Giovanna Damia
AU - Vincenza Dolo
AU - Massimo Pinzani
AU - Pier Giulio Conaldi
AU - Alessandro Bertani
AU - Lorenzo Rosso
AU - Lorenza Lazzari
AU - Vitale Miceli
PY - 2026
DA - 2026/10/02
JO - International journal of biological sciences
DO - 10.7150/ijbs.134686
UR - https://europepmc.org/article/MED/42910275
AB - Lung transplantation (LTx) remains the only curative option for patients with end-stage pulmonary diseases. However, long-term outcomes are compromised by ischemia-reperfusion injury (IRI), a major contributor to primary graft dysfunction and chronic lung allograft impairment. Current pharmacological and technological strategies provide only partial protection, highlighting the need for novel therapeutic approaches. In this study, we investigated the therapeutic potential of extracellular vesicles derived from human induced pluripotent stem cells (hiPSC-EV) in a two-dimensional (2D) in vitro model of pulmonary IRI using A549 lung epithelial cells. To better recapitulate the in vivo airway microenvironment, we also employed a human airway organoid (hAO) model. Proteomic analysis of hiPSC-EV identified 1,698 proteins enriched in pathways relevant to apoptosis regulation, oxidative stress responses, and inflammatory signaling. In the 2D IRI model, the administration of hiPSC-EV during reperfusion significantly improved cell viability, reduced lactate dehydrogenase (LDH) release, attenuated reactive oxygen species (ROS) accumulation, and decreased apoptosis and necrosis. Furthermore, hiPSC-EV downregulated pro-inflammatory mediators (NFkB1, TNFα, IL1β, IL8, MCP1) while upregulating anti-inflammatory and anti-apoptotic key factors (IL1RA, BCL2). In the 3D model, hiPSC-EV administration preserved organoid structure and viability under IRI conditions, attenuated transcriptional and protein-level expression of pro-inflammatory and oxidative stress markers (IL1β, IL8, HMGB1, HMOX1), and enhanced anti-inflammatory (IL10) and cytoprotective (HSP27, HSP70) responses. Immunofluorescence further revealed that hiPSC-EV reduced activation of apoptotic and pyroptotic pathways. Moreover, intravenous administration of hiPSC-EV in mice showed no adverse safety signals, including tumorigenicity. Together, these findings demonstrate that hiPSC-EV exert broad cytoprotective effects in pulmonary IRI through coordinated regulation of inflammation, oxidative stress, and cell death. Their scalability, reproducibility, and favorable safety profile underscore their promise as innovative cell-free therapeutics to improve outcomes in LTx.
SN - 1449-2288
ER -