A local drug delivery system prolongs graft survival by dampening T cell infiltration and neutrophil extracellular trap formation in vascularized composite allografts.

Arenas Hoyos, Isabel; Helmer, Anja; Yerly, Anaïs; Lese, Ioana; Hirsiger, Stefanie; Zhang, Lei; Casoni, Daniela; Garcia, Luisana; Petrucci, Mariafrancesca; Hammer, Sabine E; Duckova, Tereza; Banz, Yara; Montani, Matteo; Constantinescu, Mihai; Vögelin, Esther; Bordon, Gregor; Aleandri, Simone; Prost, Jean-Christophe; Taddeo, Adriano; Luciani, Paola; ... (2024). A local drug delivery system prolongs graft survival by dampening T cell infiltration and neutrophil extracellular trap formation in vascularized composite allografts. Frontiers in immunology, 15(1387945) Frontiers Research Foundation 10.3389/fimmu.2024.1387945

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INTRODUCTION

The standard treatment for preventing rejection in vascularized composite allotransplantation (VCA) currently relies on systemic immunosuppression, which exposes the host to well-known side effects. Locally administered immunosuppression strategies have shown promising results to bypass this hurdle. Nevertheless, their progress has been slow, partially attributed to a limited understanding of the essential mechanisms underlying graft rejection. Recent discoveries highlight the crucial involvement of innate immune components, such as neutrophil extracellular traps (NETs), in organ transplantation. Here we aimed to prolong graft survival through a tacrolimus-based drug delivery system and to understand the role of NETs in VCA graft rejection.

METHODS

To prevent off-target toxicity and promote graft survival, we tested a locally administered tacrolimus-loaded on-demand drug delivery system (TGMS-TAC) in a multiple MHC-mismatched porcine VCA model. Off-target toxicity was assessed in tissue and blood. Graft rejection was evaluated macroscopically while the complement system, T cells, neutrophils and NETs were analyzed in graft tissues by immunofluorescence and/or western blot. Plasmatic levels of inflammatory cytokines were measured using a Luminex magnetic-bead porcine panel, and NETs were measured in plasma and tissue using DNA-MPO ELISA. Lastly, to evaluate the effect of tacrolimus on NET formation, NETs were induced in-vitro in porcine and human peripheral neutrophils following incubation with tacrolimus.

RESULTS

Repeated intra-graft administrations of TGMS-TAC minimized systemic toxicity and prolonged graft survival. Nevertheless, signs of rejection were observed at endpoint. Systemically, there were no increases in cytokine levels, complement anaphylatoxins, T-cell subpopulations, or neutrophils during rejection. Yet, tissue analysis showed local infiltration of T cells and neutrophils, together with neutrophil extracellular traps (NETs) in rejected grafts. Interestingly, intra-graft administration of tacrolimus contributed to a reduction in both T-cellular infiltration and NETs. In fact, in-vitro NETosis assessment showed a 62-84% reduction in NETs after stimulated neutrophils were treated with tacrolimus.

CONCLUSION

Our data indicate that the proposed local delivery of immunosuppression avoids off-target toxicity while prolonging graft survival in a multiple MHC-mismatch VCA model. Furthermore, NETs are found to play a role in graft rejection and could therefore be a potential innovative therapeutic target.

Item Type:

Journal Article (Original Article)

Division/Institute:

04 Faculty of Medicine > Department of Haematology, Oncology, Infectious Diseases, Laboratory Medicine and Hospital Pharmacy (DOLS) > Institute of Clinical Chemistry
04 Faculty of Medicine > Department of Orthopaedic, Plastic and Hand Surgery (DOPH) > Clinic of Plastic and Hand Surgery > Plastic, Reconstructive and Aesthetic Surgery
04 Faculty of Medicine > Department of Orthopaedic, Plastic and Hand Surgery (DOPH) > Clinic of Plastic and Hand Surgery > Hand Surgery
04 Faculty of Medicine > Pre-clinic Human Medicine > BioMedical Research (DBMR)
04 Faculty of Medicine > Service Sector > Institute of Pathology > Clinical Pathology
04 Faculty of Medicine > Service Sector > Institute of Pathology
08 Faculty of Science > Department of Chemistry, Biochemistry and Pharmaceutical Sciences (DCBP)
04 Faculty of Medicine > Department of Orthopaedic, Plastic and Hand Surgery (DOPH) > Clinic of Plastic and Hand Surgery
04 Faculty of Medicine > Pre-clinic Human Medicine > BioMedical Research (DBMR) > Forschungsbereich Mu50 > Forschungsgruppe Handchirurgie
04 Faculty of Medicine > Pre-clinic Human Medicine > BioMedical Research (DBMR) > Forschungsbereich Mu50

UniBE Contributor:

Arenas Hoyos, Isabel, Helmer, Anja Sophie, Yerly, Anaïs Elodie, Lese, Ioana, Hirsiger, Stefanie Caroline, Zhang, Lei, Casoni, Daniela, Garcia Casalta, Luisana Gisela, Petrucci, Mariafrancesca, Banz Wälti, Yara Sarah, Montani, Matteo, Constantinescu, Mihai Adrian, Vögelin, Esther, Bordon, Gregor, Aleandri, Simone, Prost, Jean-Christophe, Taddeo, Adriano, Luciani, Paola, Rieben, Robert, Sorvillo, Nicoletta, Olariu, Radu

Subjects:

600 Technology > 610 Medicine & health
500 Science > 570 Life sciences; biology
500 Science > 540 Chemistry

ISSN:

1664-3224

Publisher:

Frontiers Research Foundation

Language:

English

Submitter:

Pubmed Import

Date Deposited:

19 Jun 2024 10:19

Last Modified:

20 Jun 2024 06:01

Publisher DOI:

10.3389/fimmu.2024.1387945

PubMed ID:

38887281

Uncontrolled Keywords:

calcineurin inhibitors (CNIs) drug delivery systems (DDSs) local immunosuppression neutrophil extracellular traps (NETs) porcine model tacrolimus transplantation immunology vascularized composite allotransplantation (VCA)

BORIS DOI:

10.48350/197919

URI:

https://boris.unibe.ch/id/eprint/197919

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