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Growth hormone receptor knockout to reduce the size of donor pigs for preclinical xenotransplantation studies
Author(s) -
Hinrichs Arne,
Riedel Evamaria O.,
Klymiuk Nikolai,
Blutke Andreas,
Kemter Elisabeth,
Längin Matthias,
Dahlhoff Maik,
Keßler Barbara,
Kurome Mayuko,
Zakhartchenko Valeri,
Jemiller EvaMaria,
Ayares David,
Bidlingmaier Martin,
Flenkenthaler Florian,
Hrabĕ de Angelis Martin,
Arnold Georg J.,
Reichart Bruno,
Fröhlich Thomas,
Wolf Eckhard
Publication year - 2020
Publication title -
xenotransplantation
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.052
H-Index - 61
eISSN - 1399-3089
pISSN - 0908-665X
DOI - 10.1111/xen.12664
Subject(s) - growth hormone receptor , xenotransplantation , offspring , biology , transgene , transplantation , andrology , growth factor , medicine , endocrinology , receptor , hormone , growth hormone , genetics , gene , pregnancy
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.