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

Autor: Hinrichs A; Department of Veterinary Sciences, Chair for Molecular Animal Breeding and Biotechnology, Gene Center, LMU Munich, Munich, Germany.; Center for Innovative Medical Models (CiMM), LMU Munich, Munich, Germany., Riedel EO; Laboratory for Functional Genome Analysis (LAFUGA), Gene Center, LMU Munich, Munich, Germany., Klymiuk N; Department of Veterinary Sciences, Chair for Molecular Animal Breeding and Biotechnology, Gene Center, LMU Munich, Munich, Germany.; Center for Innovative Medical Models (CiMM), LMU Munich, Munich, Germany., Blutke A; Institute of Experimental Genetics, Helmholtz Zentrum München, Chair of Experimental Genetics, Technical University of Munich, Neuherberg, Germany., Kemter E; Department of Veterinary Sciences, Chair for Molecular Animal Breeding and Biotechnology, Gene Center, LMU Munich, Munich, Germany.; Center for Innovative Medical Models (CiMM), LMU Munich, Munich, Germany., Längin M; Department of Anaesthesiology, University Hospital, LMU Munich, Munich, Germany., Dahlhoff M; Department of Veterinary Sciences, Chair for Molecular Animal Breeding and Biotechnology, Gene Center, LMU Munich, Munich, Germany., Keßler B; Department of Veterinary Sciences, Chair for Molecular Animal Breeding and Biotechnology, Gene Center, LMU Munich, Munich, Germany.; Center for Innovative Medical Models (CiMM), LMU Munich, Munich, Germany., Kurome M; Department of Veterinary Sciences, Chair for Molecular Animal Breeding and Biotechnology, Gene Center, LMU Munich, Munich, Germany.; Center for Innovative Medical Models (CiMM), LMU Munich, Munich, Germany., Zakhartchenko V; Department of Veterinary Sciences, Chair for Molecular Animal Breeding and Biotechnology, Gene Center, LMU Munich, Munich, Germany.; Center for Innovative Medical Models (CiMM), LMU Munich, Munich, Germany., Jemiller EM; Department of Veterinary Sciences, Chair for Molecular Animal Breeding and Biotechnology, Gene Center, LMU Munich, Munich, Germany.; Center for Innovative Medical Models (CiMM), LMU Munich, Munich, Germany., Ayares D; Revivicor Inc., Blacksburg, VA, USA., Bidlingmaier M; Medizinische Klinik und Poliklinik IV, Klinikum der Universität München, Munich, Germany., Flenkenthaler F; Laboratory for Functional Genome Analysis (LAFUGA), Gene Center, LMU Munich, Munich, Germany., Hrabĕ de Angelis M; Institute of Experimental Genetics, Helmholtz Zentrum München, Chair of Experimental Genetics, Technical University of Munich, Neuherberg, Germany., Arnold GJ; Laboratory for Functional Genome Analysis (LAFUGA), Gene Center, LMU Munich, Munich, Germany., Reichart B; Walter Brendel Center for Experimental Medicine, LMU Munich, Munich, Germany., Fröhlich T; Laboratory for Functional Genome Analysis (LAFUGA), Gene Center, LMU Munich, Munich, Germany., Wolf E; Department of Veterinary Sciences, Chair for Molecular Animal Breeding and Biotechnology, Gene Center, LMU Munich, Munich, Germany.; Center for Innovative Medical Models (CiMM), LMU Munich, Munich, Germany.; Laboratory for Functional Genome Analysis (LAFUGA), Gene Center, LMU Munich, Munich, Germany.
Jazyk: angličtina
Zdroj: Xenotransplantation [Xenotransplantation] 2021 Mar; Vol. 28 (2), pp. e12664. Date of Electronic Publication: 2020 Nov 25.
DOI: 10.1111/xen.12664
Abstrakt: 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.
(© 2020 The Authors. Xenotransplantation published by John Wiley & Sons Ltd.)
Databáze: MEDLINE
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