Genetically engineered pigs and target-specific immunomodulation provide significant graft survival and hope for clinical cardiac xenotransplantation

Mohiuddin, Muhammad M.; Singh, Avneesh K.; Corcoran, Philip C.; Hoyt, Robert F.; Thomas, Marvin L.; Ayares, David; Horvath, Keith A.

doi:10.1016/j.jtcvs.2014.06.002

Cited by 112 publications

(123 citation statements)

References 23 publications

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“…After producing hDAF transgenic and α1,3-galactosyltransferase (α1,3GT) knockout (KO) pigs, hyperacute rejection (HAR) [1,2] no longer appeared to be a problem [3]. However, acute humoral xenograft rejection (AHXR) [4] is defined as a rejection that begins within 24 h after transplantation and gradually destroys the graft. The origin of AHXR continues to be a controversial topic, but non-Gal antigen-antibody reactions and thrombotic microangiopathy are included in this definition.…”

Section: Introductionmentioning

confidence: 99%

Suppression of human macrophage-mediated cytotoxicity by transgenic swine endothelial cell expression of HLA-G

Esquivel

Maeda

Eguchi

et al. 2015

Transplant Immunology

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Section: Introductionmentioning

confidence: 99%

Suppression of human macrophage-mediated cytotoxicity by transgenic swine endothelial cell expression of HLA-G

Esquivel

Maeda

Eguchi

et al. 2015

Transplant Immunology

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“…The pig has been a primary candidate donor, but the expression of a sugar molecule that induces hyperrejection in humans, as well as complement activation and coagulopathies, has interfered with progress until recently. Employing pigs genetically engineered to reduce these problems, investigators at the National Heart, Lung, and Blood Institute were able to successfully transplant pig hearts to baboons [56,57]. Modifications to the immunosuppression protocol permitted cardiac xenograft survival of up to one year.…”

Section: Heart Muscle and Vascular Regenerationmentioning

confidence: 99%

“…Pig lungs are similar in size to human lungs, and as a result, generation of pigs engineered to reduce rejection [56,57] has rekindled interest in the use of pigs as xenogeneic lung donors; this is likely an area of future research.…”

Section: Lung Regenerationmentioning

confidence: 99%

Translational Animal Models for Regenerative Medicine Research

Sharp

2015

Translational Regenerative Medicine

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“…Several triple transgenic animals were generated and used for pig-to-baboon heterotopic/orthotopic heart transplantation to verify the positive effects of human thrombomodulin for xenotransplantation. In a heterotopic transplantation model hearts from GGTA1 knockout/CD46 animals survived between of 21 to 80 days, whereas hearts from GGTA1 knockout/CD46/human thrombomodulin animals survived on average for more than 200 days, indeed some animals surpassed one (Mohiuddin et al, 2014) and even two years (MIT Report, 2015).…”

Section: Antithrombotic Genesmentioning

confidence: 99%

“…This CD46 minigene was chosen because animals transgenic for this construct have already been intensively characterized and served as the basis for CD46-hTM double transgenic animals, which have provided the most successful pig to baboon heterotopic heart xenotransplantation so far (Mohiuddin et al, 2014).…”

Section: The Cd46 Minigenementioning

confidence: 99%

Multi‐transgenic pigs for xenotransplantation

Fischer

Kraner-Scheiber

Flisikowski

et al. 2013

Xenotransplantation

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Genetically modified pigs offer a solution to the severe shortage of organs available for human transplantation. Inactivation of the α1,3‐galactosyltransferase gene, responsible for α1,3‐Gal epitope synthesis (1–2) was a major breakthrough and essentially solved the problem of hyperacute rejection. However, further modifications of donor pigs are required to enable long term xenograft survival. These include expression of complement regulatory, antithrombotic, endothelium protective and immuno‐regulatory transgenes. Current evidence indicates that complement regulator transgenes, such as CD46 and CD55, must be expressed at least fivefold more than their human endogenous equivalent to effectively protect grafted tissue (3). To explore means of achieving high expression, we produced a series of BAC‐ and PAC‐vector constructs containing different sized genomic complement regulatory genes, from 70 to 190 kb, under the control of endogenous or CAGGs (CMV enhancer chicken β actin) promoter sequences. Using these vectors we obtained high levels of CD46 and CD55 expression in vitro. To prepare multi‐transgene “packages” for introduction into animals, the most effective constructs were combined with one or two further xenoprotective transgenes, including A20 (4), HO‐1 (5), thrombomodulin (6) and CTLA4‐Ig (LEA 29Y). In cotransfection experiments single cell clones expressing up to five different xenogenes could be detected. Achieving desired levels of transgene expression in an animal is not only a function of the transgene construct, but also of the location at which it integrates. Position effect variation in expression of random transgenes is well‐documented. Transgene placement by gene targeting at a permissive locus offers a useful means of achieving reliable transgene expression. In mice the ROSA26 locus is widely used to support ubiquitous expression of inserted transgenes (7–8). Murine ROSA26 encodes no functional genes and extensive gene targeting of this locus has not led to deleterious effects on development or physiology (9). To enable a similar approach in pigs, we recently identified a strong candidate for the porcine ROSA26 locus. Gene targeted placement of a neomycin reporter gene construct under the control of the ROSA26 promoter showed expression in all examined porcine tissues of all three germ layers. This strongly suggests that porcine ROSA 26 may resemble the murine locus as a suitably permissive site for transgene expression. We are currently proceeding with gene targeting to place various xenoprotective transgene constructs at this locus. References: 1. Dai Y, Vaught TD, Boone J et al. Targeted disruption of the alpha1,3‐ galactosyltransferase gene in cloned pigs. Nat Biotechnol 2002; 20: 251–255. 2. Lai L, Kolber‐Simonds D, Park K et al. Production of alpha‐1,3‐galactosyltransferase knockout pigs by nuclear transfer cloning. Science 2002; 295: 1089–1092. 3. Miyagawa S, Fukuta D, Kitano E et al. Effect of tandem forms of DAF(CD55) on complement‐mediated xenogeneic cell lysis. Xenotransplantation 20...

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Genetically engineered pigs and target-specific immunomodulation provide significant graft survival and hope for clinical cardiac xenotransplantation

Cited by 112 publications

References 23 publications

Suppression of human macrophage-mediated cytotoxicity by transgenic swine endothelial cell expression of HLA-G

Suppression of human macrophage-mediated cytotoxicity by transgenic swine endothelial cell expression of HLA-G

Translational Animal Models for Regenerative Medicine Research

Multi‐transgenic pigs for xenotransplantation

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