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Diabetologia

, Volume 61, Issue 8, pp 1804–1810 | Cite as

Vegf-A mRNA transfection as a novel approach to improve mouse and human islet graft revascularisation

  • Willem Staels
  • Yannick Verdonck
  • Yves Heremans
  • Gunter Leuckx
  • Sofie De Groef
  • Carlo Heirman
  • Eelco de Koning
  • Conny Gysemans
  • Kris Thielemans
  • Luc Baeyens
  • Harry Heimberg
  • Nico De Leu
Short Communication

Abstract

Aims/hypothesis

The initial avascular period following islet transplantation seriously compromises graft function and survival. Enhancing graft revascularisation to improve engraftment has been attempted through virus-based delivery of angiogenic triggers, but risks associated with viral vectors have hampered clinical translation. In vitro transcribed mRNA transfection circumvents these risks and may be used for improving islet engraftment.

Methods

Mouse and human pancreatic islet cells were transfected with mRNA encoding the angiogenic growth factor vascular endothelial growth factor A (VEGF-A) before transplantation under the kidney capsule in mice.

Results

At day 7 post transplantation, revascularisation of grafts transfected with Vegf-A (also known as Vegfa) mRNA was significantly higher compared with non-transfected or Gfp mRNA-transfected controls in mouse islet grafts (2.11- and 1.87-fold, respectively) (vessel area/graft area, mean ± SEM: 0.118 ± 0.01 [n = 3] in Vegf-A mRNA transfected group (VEGF) vs 0.056 ± 0.01 [n = 3] in no RNA [p < 0.05] vs 0.063 ± 0.02 [n = 4] in Gfp mRNA transfected group (GFP) [p < 0.05]); EndoC-bH3 grafts (2.85- and 2.48-fold. respectively) (0.085 ± 0.02 [n = 4] in VEGF vs 0.030 ± 0.004 [n = 4] in no RNA [p < 0.05] vs 0.034 ± 0.01 [n = 5] in GFP [p < 0.05]); and human islet grafts (3.17- and 3.80-fold, respectively) (0.048 ± 0.013 [n = 3] in VEGF vs 0.015 ± 0.0051 [n = 4] in no RNA [p < 0.01] vs 0.013 ± 0.0046 [n = 4] in GFP [p < 0.01]). At day 30 post transplantation, human islet grafts maintained a vascularisation benefit (1.70- and 1.82-fold, respectively) (0.049 ± 0.0042 [n = 8] in VEGF vs 0.029 ± 0.0052 [n = 5] in no RNA [p < 0.05] vs 0.027 ± 0.0056 [n = 4] in GFP [p < 0.05]) and a higher beta cell volume (1.64- and 2.26-fold, respectively) (0.0292 ± 0.0032 μl [n = 7] in VEGF vs 0.0178 ± 0.0021 μl [n = 5] in no RNA [p < 0.01] vs 0.0129 ± 0.0012 μl [n = 4] in GFP [p < 0.001]).

Conclusions/interpretation

Vegf-A mRNA transfection before transplantation provides a promising and safe strategy to improve engraftment of islets and other cell-based implants.

Keywords

Cell therapy Diabetes Gene delivery Graft revascularisation Islet transplantation Messenger RNA Pancreatic beta cell RNA delivery VEGFA 

Abbreviations

GFP

Gfp mRNA transfected group

IVT

In vitro transcription

modVEGF

modified Vegf-A mRNA transfected group

SCID

Severe combined immunodeficiency

VEGF

Vegf-A mRNA transfected group

VEGF-A

Vascular endothelial growth factor A

Notes

Acknowledgements

The authors thank V. Laurysens, A. Demarré and E. Quartier (BENE, Vrije Universiteit Brussel, Brussels, Belgium) for technical help. We also thank J. van den Ameele (Andrea Brand Lab, Gurdon Institute, University of Cambridge, Cambridge, UK) for revising the manuscript.

Contribution statement

WS, EdK, CG, CH, KT, LB, HH and NDL designed and conceived the experiments. WS, YV, YH, GL and SDG acquired and analysed the data. EdK provided human islets. CH and KT provided Gfp mRNA, Vegf-A mRNA and modified Vegf-A mRNA. WS, YV, YH, CH, KT, CG, LB, HH and NDL interpreted the data. WS and NDL drafted the article. All authors revised the article and approved of the final version. NDL is the guarantor of this work.

Funding

The authors acknowledge support by grants from the Research Foundation Flanders (FWO), the VUB Research Council, Stichting Diabetes Onderzoek Nederland, the European Union Sixth and Seventh Framework Program, the Wetenschappelijk Fonds Willy Gepts (WFWG) of the UZ Brussel and the Belgian Federal Science Policy (IAPVII-07). WS is a PhD fellow of Research Foundation Flanders.

Duality of interest

The authors declare that there is no duality of interest associated with this manuscript.

Supplementary material

125_2018_4646_MOESM1_ESM.pdf (4.1 mb)
ESM (PDF 4.06 mb)

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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Beta Cell Neogenesis (BENE), Vrije Universiteit BrusselBrusselsBelgium
  2. 2.Department of Paediatrics, Division of Paediatric EndocrinologyGhent UniversityGhentBelgium
  3. 3.Laboratory of Molecular and Cellular TherapyVrije Universiteit BrusselBrusselsBelgium
  4. 4.Department of Medicine, Section of EndocrinologyLeiden University Medical CenterLeidenthe Netherlands
  5. 5.Laboratory of Clinical and Experimental EndocrinologyKatholieke Universiteit LeuvenLeuvenBelgium
  6. 6.Department of EndocrinologyUZ BrusselBrusselsBelgium
  7. 7.Department of EndocrinologyASZ AalstAalstBelgium

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