MTOR Inhibitor Everolimus in Regulatory T Cell Expansion for Clinical Application in Transplantation. Issue 4 (April 2019)

• Record Type: Journal Article
• Authors: Gedaly, Roberto; De Stefano, Felice; Turcios, Lilia; Hill, Marita; Hidalgo, Giovanna; Mitov, Mihail I.; Alstott, Michael C.; Butterfield, D. Allan; Mitchell, Hunter C.; Hart, Jeremy; Al-Attar, Ahmad; Jennings, Chester D.; Marti, Francesc
• Is Part Of: Transplantation. Volume 103:Issue 4(2019)
• Publication Date: 2019-04
• Subjects: Transplantation of organs, tissues, etc -- Periodicals; Transplantation immunology -- Periodicals; 617.95
• DOI: 10.1097/TP.0000000000002495
• Languages: English
• ISSNs: 0041-1337

Abstract

Abstract : Background: Experimental and preclinical evidence suggest that adoptive transfer of regulatory T (Treg) cells could be an appropriate therapeutic strategy to induce tolerance and improve graft survival in transplanted patients. The University of Kentucky Transplant Service Line is developing a novel phase I/II clinical trial with ex vivo expanded autologous Treg cells as an adoptive cellular therapy in renal transplant recipients who are using everolimus (EVR)-based immunosuppressive regimen. Methods: The aim of this study was to determine the mechanisms of action and efficacy of EVR for the development of functionally competent Treg cell-based adoptive immunotherapy in transplantation to integrate a common EVR-based regimen in vivo (in the patient) and ex vivo (in the expansion of autologous Treg cells). CD25 + Treg cells were selected from leukapheresis product with a GMP-compliant cell separation system and placed in 5-day (short) or 21-day (long) culture with EVR or rapamycin (RAPA). Multi-parametric flow cytometry analyses were used to monitor the expansion rates, phenotype, autophagic flux, and suppressor function of the cells. phosphoinositide 3-kinase/protein kinase B/mammalian target of rapamycin signaling pathway profiles of treated cells were analyzed by Western blot and cell bioenergetic parameters by extracellular flux analysis. Results: EVR-treated cells showed temporary slower growth, lower metabolic rates, and reduced phosphorylation of protein kinase B compared with RAPA-treated cells. In spite of these differences, the expansion rates, phenotype, and suppressor function of long-term Treg cells in culture with EVR were similar to those with RAPA. Conclusions: Our results support the feasibility of EVR to expand functionally competent Treg cells for their clinical use. Abstract : Supplemental Digital Content is available in the text.