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Gentherapie der Transplantatvaskulopathie

Gene therapy for transplant vasculopathy

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Zeitschrift für Herz-,Thorax- und Gefäßchirurgie Aims and scope

Zusammenfassung

Das Forschungsprojekt befasste sich mit der gentherapeutischen Behandlung der Transplantatvaskulopathie (TVP). Unter Verwendung des heterotopen Aortentransplantationsmodells der Maus konnte ein suffizientes Modell etabliert werden, das im translationalen Sinne die Ex-vivo-Behandlung der zu transplantierenden Aortenfragmente erlaubt. Diesbezüglich wurden drei Ansätze verfolgt: 1) Durch die Neutralisierung des Transkriptionsfaktors Aktivatorprotein 1 (AP-1) mit einem DNA-Haarnadel-Molekül (Decoy-Oligodesoxynukleotid, dODN) konnte nach 30 Tagen eine signifikante Reduktion der Intimahyperplasie nachgewiesen werden. Die Migration glatter Muskelzellen und die Expression von Matrix-Metalloproteinasen (MMP) werden u. a. durch AP‑1 induziert. 2) Es folgte die Implementierung eines RNA-AP-1-Decoy-Oligonukleotids (dON), das nach Transduktion der Transplantate mit einem nichtpathogenen adenoassoziierten Virus (AAV) langfristig in den Zellen exprimiert wurde. In diesem Ansatz konnte erneut eine signifikante Reduktion der Intimahyperplasie und eine signifikante Reduktion von VCAM‑1, Makrophageninfiltration und von CD4-positiven Zellen sowie eine reduzierte MMP-9-Aktivität nachweisen werden. 3) Durch die AAV-vermittelte Überexpression des natürlichen MMP-Inhibitors TIMP‑1 gelang es, die MMP-Aktivität zu hemmen und dadurch die TVP zu reduzieren. Zudem konnten eine verbesserte Barriereeigenschaft des Endothels sowohl in vitro als auch in vivo nachgewiesen werden.

Abstract

This research project addressed the gene therapy treatment of transplant vasculopathy (TVP). Using the heterotopic mouse aortic transplantation model, a sufficient model was established that allowed to treat in a translational sense the aortic fragments to be transplanted ex vivo. In this respect three approaches followed: 1) by neutralizing the transcription factor activator protein 1 (AP-1) with a DNA hairpin molecule (decoy oligodeoxynucleotide, dODN), a significant reduction of intimal hyperplasia after 30 days was demonstrated. AP‑1 is a transcription factor that induces smooth muscle cell migration and expression of matrix metalloproteinases (MMP), among others. 2) It followed the development of an RNA AP‑1 decoy oligonucleotide (dON) that showed long-term expression in the cells after the transduction of the grafts with a nonpathogenic adeno-associated virus (AAV). In this approach, again a significant reduction in intimal hyperplasia and a significant reduction in VCAM‑1, macrophage infiltration, and CD4 positive cells as well as reduced MMP‑9 activity were demonstrated. 3) Through AAV-mediated overexpression of the natural MMP inhibitor TIMP‑1, it was possible to inhibit MMP activity and thereby reduce TVP. In addition, an improved barrier function of endothelial cells in vitro and in vivo was demonstrated.

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Förderung

Das Projekt wurde von der Dietmar-Hopp-Stiftung (Projektnummer 23011198) gefördert.

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Authors and Affiliations

  1. Klinik für Herzchirurgie, Universitätsklinikum Heidelberg, Heidelberg, Deutschland

    Rawa Arif

  2. Herzchirurgie, INCCI HaerzZenter, Luxemburg, Luxemburg

    Klaus Kallenbach

  3. Klinik für Innere Medizin III (Kardiologie, Angiologie und Internistische Intensivmedizin), Universitätsklinikum Schleswig-Holstein und Christian-Albrechts-Universität zu Kiel, Kiel, Deutschland

    Oliver J. Müller

  4. Institut für Physiologie und Pathophysiologie, Abteilung Herz- und Kreislaufphysiologie, Universität Heidelberg, Im Neuenheimer Feld 326, 69120, Heidelberg, Deutschland

    Andreas H. Wagner

Authors
  1. Rawa Arif
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  2. Klaus Kallenbach
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  4. Andreas H. Wagner
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Corresponding author

Correspondence to Andreas H. Wagner.

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Interessenkonflikt

R. Arif, K. Kallenbach, O.J. Müller und A.H. Wagner geben an, dass kein Interessenkonflikt besteht. Alle Autoren haben zur Entstehung des Manuskripts beigetragen und sind mit dessen Veröffentlichung einverstanden.

Alle nationalen Richtlinien zur Haltung und zum Umgang mit Labortieren wurden eingehalten, und die notwendigen Zustimmungen der zuständigen Behörden liegen vor.

Additional information

PD Dr. med. Rawa Arif wurde für seine Forschungen „Methoden zur Prävention der Transplantatvaskulopathie“ 2021 mit dem Franz J. Köhler-Preis der Deutschen Gesellschaft für Thorax‑, Herz- und Gefäßchirurgie (DGTHG) ausgezeichnet. Der Beitrag ist eine Zusammenfassung der Originalarbeiten: [12,13,14].

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Arif, R., Kallenbach, K., Müller, O.J. et al. Gentherapie der Transplantatvaskulopathie. Z Herz- Thorax- Gefäßchir 36, 328–333 (2022). https://doi.org/10.1007/s00398-022-00535-5

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  • DOI: https://doi.org/10.1007/s00398-022-00535-5

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