Exosomal-miRNas expression and growth factors released by mononuclear cells of CLAD patients in response to extracorporeal photopheresis

• Published in: Journal of translational medicine 2024-03, Vol.22 (1), p.276-10, Article 276
• Main Authors: Bozzini, Sara, Bozza, Eleonora, Bagnera, Cecilia, Del Fante, Claudia, Barone, Eugenio, De Vitis, Simona, De Amici, Mara, Testa, Giorgia, Croce, Stefania, Valsecchi, Chiara, Avanzini, Maria A, Cacciatore, Rosalia, Mortellaro, Cristina, Viarengo, Gianluca, Perotti, Cesare, Meloni, Federica
• Format: Article
• Language: English
• Subjects: Analysis; Antigens; Blood; Cell activation; Cell culture; CLAD; Connective tissue growth factor; Connective tissues; Dendritic cells; ECP; Exosomal-miRNas; Exosomes; Extracorporeal photopheresis; Flow cytometry; Growth factors; Immunoregulation; Leukapheresis; Leukocytes; Leukocytes (mononuclear); Lipopolysaccharides; Lung transplantation; Lung transplants; Lungs; Lymphocytes T; Macrophages; MicroRNA; MicroRNAs; miRNA; Molecular modelling; Patient outcomes; Patients; Quality control; Regulation; T regulatory cells; Testing; Thermal cycling; Transplantation
• ISSN: 1479-5876; 1479-5876
• DOI: 10.1186/s12967-024-05045-6

CLAD (Chronic Lung Allograft Dysfunction) remains a serious complication following lung transplantation. Some evidence shows that portions of Extracorporeal Photopheresis (ECP)-treated patients improve/stabilize their graft function. In spite of that, data concerning molecular mechanisms are still lacking. Aims of our study were to assess whether ECP effects are mediated by Mononuclear Cells (MNCs) modulation in term of microRNAs (miRNAs) expression and growth factors release. Cells from leukapheresis of 16 CLAD patients, at time 0 and 6-months (10 cycles), were cultured for 48h ± PHA (10 ug/ml) or LPS (2 ug/ml). Expression levels of miR-146a-5p, miR-155-5p, miR-31-5p, miR181a-5p, miR-142-3p, miR-16-5p and miR-23b-5p in MNCs-exosomes were evaluated by qRT-PCR, while ELISA assessed different growth factors levels on culture supernatants. Our result showed miR-142-3p down-regulation (p = 0.02) in MNCs of ECP-patients after the 10 cycles and after LPS stimulation (p = 0.005). We also find miR-146a-5p up-regulation in cells after the 10 cycles stimulated with LPS (p = 0.03). Connective tissue growth factor (CTGF) levels significantly decreased in MNCs supernatant (p = 0.04). The effect of ECP is translated into frequency changes of Dendritic Cell (DC) subpopulations and a slight increase in T regulatory cells (Treg) number and a significant decrease in CTGF release. ECP might affect regulatory T cell functions, since both miR-142 and miR-146a have been shown to be involved in the regulation of suppressor regulatory T cell functions and DCs. On the other side ECP, possibly by regulating macrophage activation, is able to significantly down modulate CTGF release.