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. 2021 Mar;28(2):e12664.
doi: 10.1111/xen.12664. Epub 2020 Nov 25.

Growth hormone receptor knockout to reduce the size of donor pigs for preclinical xenotransplantation studies

Arne Hinrichs  1   2 , Evamaria O Riedel  3 , Nikolai Klymiuk  1   2 , Andreas Blutke  4 , Elisabeth Kemter  1   2 , Matthias Längin  5 , Maik Dahlhoff  1 , Barbara Keßler  1   2 , Mayuko Kurome  1   2 , Valeri Zakhartchenko  1   2 , Eva-Maria Jemiller  1   2 , David Ayares  6 , Martin Bidlingmaier  7 , Florian Flenkenthaler  3 , Martin Hrabĕ de Angelis  4 , Georg J Arnold  3 , Bruno Reichart  8 , Thomas Fröhlich  3 , Eckhard Wolf  1   2   3
Affiliations

Growth hormone receptor knockout to reduce the size of donor pigs for preclinical xenotransplantation studies

Arne Hinrichs et al. Xenotransplantation. 2021 Mar.

Abstract

Background: Many genetically multi-modified donor lines for xenotransplantation have a background of domestic pigs with rapid body and organ growth. The intrinsic growth potential of porcine xeno-organs may impair their long-term function after orthotopic transplantation in non-human primate models. Since growth hormone is a major stimulator of postnatal growth, we deleted its receptor (GHR-KO) to reduce the size of donor pigs in one step.

Methods: Heart weight and proteome profile of myocardium were investigated in GHR-KO and control pigs. GHR-KO mutations were introduced using CRISPR/Cas9 in an α1,3-galactosyltransferase (GGTA1)-deficient background expressing the human cluster of differentiation (hCD46) and human thrombomodulin (hTHBD) to generate quadruple-modified (4GM) pigs.

Results: At age 6 months, GHR-KO pigs had a 61% reduced body weight and a 63% reduced heart weight compared with controls. The mean minimal diameter of cardiomyocytes was 28% reduced. A holistic proteome study of myocardium samples from the two groups did not reveal prominent differences. Two 4GM founder sows had low serum insulin-like growth factor 1 (IGF1) levels (24 ± 1 ng/mL) and reached body weights of 70.3 and 73.4 kg at 9 months. Control pigs with IGF1 levels of 228 ± 24 ng/mL reached this weight range three months earlier. The 4GM sows showed normal sexual development and were mated with genetically multi-modified boars. Offspring revealed the expected Mendelian transmission of the genetic modifications and consistent expression of the transgenes.

Conclusion: GHR-KO donor pigs can be used at an age beyond the steepest phase of their growth curve, potentially reducing the problem of xeno-organ overgrowth in preclinical studies.

Keywords: gene editing; growth hormone receptor; organ growth; pig.

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References

REFERENCES

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