Author Correction: Consistent success in life-supporting porcine cardiac xenotransplantation

Längin, Matthias; Mayr, Tanja; Reichart, B.; Michel, Sebastian; Buchholz, Stefan; Guethoff, Sonja; Dashkevich, Alexey; Baehr, Andrea; Egerer, Stefanie; Bauer, Andreas; Mihalj, Maks; Panelli, Alessandro; Issl, Lara; Ying, Jiawei; Fresch, Ann Kathrin; Buttgereit, Ines; Mokelke, Maren; Radan, Julia; Werner, Fabian; Lutzmann, I.; Steen, Stig; Sjöberg, Trygve; Paskevicius, Audrius; Liao, Qiuming; Sfriso, Riccardo; Rieben, Robert; Dahlhoff, Maik; Keßler, Barbara; Kemter, Elisabeth; Kurome, Mayuko; Zakhartchenko, Valeri; Klett, Katharina; Hinkel, Rabea; Kupatt, Christian; Falkenau, Almuth; Reu, Simone; Ellgass, Reinhard; Herzog, Rudolf; Binder, Uli; Wich, Günter; Skerra, Arne; Ayares, David; Kind, Alexander; Schönmann, Uwe; Kaup, Franz‐Josef; Hagl, Christian; Wolf, Eckhard; Klymiuk, Nikolai; Brenner, Paolo; Abicht, Jan‐Michael

doi:10.1038/s41586-019-1108-4

Cited by 6 publications

(3 citation statements)

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“…Transgenic pigs for complement regulatory protein such as hCD46, hCD55, and hCD59 were developed to overcome complement-mediated cytotoxicity or early graft failure compared to wild-type pigs or GTKO pigs 5 , 14 – 17 . GTKO/hCD46/hTBM pigs showed the longest survival (945 days for heterotopic, 195 days for orthotopic) in cardiac pig-to-primate xenografts 18 , 19 .…”

Section: Introductionmentioning

confidence: 98%

A desirable transgenic strategy using GGTA1 endogenous promoter-mediated knock-in for xenotransplantation model

Shim

Kim

et al. 2022

Sci Rep

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Pig-to-human organ transplantation is a feasible solution to resolve the shortage of organ donors for patients that wait for transplantation. To overcome immunological rejection, which is the main hurdle in pig-to-human xenotransplantation, various engineered transgenic pigs have been developed. Ablation of xeno-reactive antigens, especially the 1,3-Gal epitope (GalT), which causes hyperacute rejection, and insertion of complement regulatory protein genes, such as hCD46, hCD55, and hCD59, and genes to regulate the coagulation pathway or immune cell-mediated rejection may be required for an ideal xenotransplantation model. However, the technique for stable and efficient expression of multi-transgenes has not yet been settled to develop a suitable xenotransplantation model. To develop a stable and efficient transgenic system, we knocked-in internal ribosome entry sites (IRES)-mediated transgenes into the α 1,3-galactosyltransferase (GGTA1) locus so that expression of these transgenes would be controlled by the GGTA1 endogenous promoter. We constructed an IRES-based polycistronic hCD55/hCD39 knock-in vector to target exon4 of the GGTA1 gene. The hCD55/hCD39 knock-in vector and CRISPR/Cas9 to target exon4 of the GGTA1 gene were co-transfected into white yucatan miniature pig fibroblasts. After transfection, hCD39 expressed cells were sorted by FACS. Targeted colonies were verified using targeting PCR and FACS analysis, and used as donors for somatic cell nuclear transfer. Expression of GalT, hCD55, and hCD39 was analyzed by FACS and western blotting. Human complement-mediated cytotoxicity and human antibody binding assays were conducted on peripheral blood mononuclear cells (PBMCs) and red blood cells (RBCs), and deposition of C3 by incubation with human complement serum and platelet aggregation were analyzed in GGTA1 knock-out (GTKO)/CD55/CD39 pig cells. We obtained six targeted colonies with high efficiency of targeting (42.8% of efficiency). Selected colony and transgenic pigs showed abundant expression of targeted genes (hCD55 and hCD39). Knocked-in transgenes were expressed in various cell types under the control of the GGTA1 endogenous promoter in GTKO/CD55/CD39 pig and IRES was sufficient to express downstream expression of the transgene. Human IgG and IgM binding decreased in GTKO/CD55/CD39 pig and GTKO compared to wild-type pig PBMCs and RBCs. The human complement-mediated cytotoxicity of RBCs and PBMCs decreased in GTKO/CD55/CD39 pig compared to cells from GTKO pig. C3 was also deposited less in GTKO/CD55/CD39 pig cells than wild-type pig cells. The platelet aggregation was delayed by hCD39 expression in GTKO/CD55/CD39 pig. In the current study, knock-in into the GGTA1 locus and GGTA1 endogenous promoter-mediated expression of transgenes are an appropriable strategy for effective and stable expression of multi-transgenes. The IRES-based polycistronic transgene vector system also caused sufficient expression of both hCD55 and hCD39. Furthermore, co-transfection of CRISPR/Cas9 and the knock-in vector not only increased the knock-in efficiency but also induced null for GalT by CRISPR/Cas9-mediated double-stranded break of the target site. As shown in human complement-mediated lysis and human antibody binding to GTKO/CD55/CD39 transgenic pig cells, expression of hCD55 and hCD39 with ablation of GalT prevents an effective immunological reaction in vitro. As a consequence, our technique to produce multi-transgenic pigs could improve the development of a suitable xenotransplantation model, and the GTKO/CD55/CD39 pig developed could prolong the survival of pig-to-primate xenotransplant recipients.

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Section: Introductionmentioning

confidence: 98%

A desirable transgenic strategy using GGTA1 endogenous promoter-mediated knock-in for xenotransplantation model

Shim

Kim

et al. 2022

Sci Rep

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“… 8 Research using an organ from these modified animals transplanted in non-human primates accomplished promising results, especially in terms of survival and function. 9 …”

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confidence: 99%

Bioethics and xenotransplantation from pig to human

Galvão

Grinberg

2023

Clinics

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“…The experiments of kidney, heart, and liver xenotransplantation were attempted without lasting success [ 2 ]. After a few decades of investigation, significant breakthroughs and preclinical milestones have been achieved worldwide [ 3 , 4 , 5 ], yet biological barriers and ethical issues have long been preventing the clinical trial of xenotransplantation from being carried out, therefore delaying the clinical application of xenotransplantation in the foreseeable future [ 6 , 7 ].…”

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confidence: 99%

Expert consensus on clinical trials of human xenotransplantation in China

Huang¹

2022

Health Care Science

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The history of xenotransplantation started in the 19th century. After a few decades of investigation, significant breakthroughs and preclinical milestones have been achieved worldwide. With the recent transplantation of genetically modified porcine kidneys and heart into humans, these ground-breaking achievements have attracted great attention worldwide, in the hope that xenotransplantation might alleviate or even solve the problem of organ shortage. On January 20, 2022, the China Organ Transplantation Development Foundation convened a symposium on "The History, Current Situation and Future of Human Xenotransplantation Clinical Trials," where ways to promote the ethical and sustainable development of xenotransplantation in China were discussed among the participating experts. A formal consensus was reached as the product of the symposium, outlining the expert opinions on scientific, regulatory, and ethical issues of clinical trials of xenotransplantation in China.

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Author Correction: Consistent success in life-supporting porcine cardiac xenotransplantation

Cited by 6 publications

References 0 publications

A desirable transgenic strategy using GGTA1 endogenous promoter-mediated knock-in for xenotransplantation model

A desirable transgenic strategy using GGTA1 endogenous promoter-mediated knock-in for xenotransplantation model

Bioethics and xenotransplantation from pig to human

Expert consensus on clinical trials of human xenotransplantation in China

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