Antigenicity of tissues and organs from GGTA1/CMAH4GalNT2 triple gene knockout pigs

Ronggen Wang, Miaomiao Ruan, Runjie Zhang, Lei Chen, Xiaoxue Li, Bin Fang, Chu Li, Xueyang Ren, Jiying Liu, Qiang Xiong, Lining Zhang, Yong Jin, Lin Li, Rongfeng Li, Ying Wang, Haiyuan Yang, Yifan Dai

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Journal of Biomedical Research ›› DOI: 10.7555/JBR.32.20180018
Original Article

Antigenicity of tissues and organs from GGTA1/CMAH4GalNT2 triple gene knockout pigs

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Abstract

Clinical xenotransplantations have been hampered by human preformed antibody-mediated damage of the xenografts. To overcome biological incompatibility between pigs and humans, one strategy is to remove the major antigens [Gal, Neu5Gc, and Sd(a)] present on pig cells and tissues. Triple gene (GGTA1, CMAH, and b4GalNT2) knockout (TKO) pigs were produced in our laboratory by CRISPR-Cas9 targeting. To investigate the antigenicity reduction in the TKO pigs, the expression levels of these three xenoantigens in the cornea, heart, liver, spleen, lung, kidney, and pancreas tissues were examined. The level of human IgG/IgM binding to those tissues was also investigated, with wildtype pig tissues as control. The results showed that αGal, Neu5Gc, and Sd(a) were markedly positive in all the examined tissues in wildtype pigs but barely detected in TKO pigs. Compared to wildtype pigs, the liver, spleen, and pancreas of TKO pigs showed comparable levels of human IgG and IgM binding, whereas corneas, heart, lung, and kidney of TKO pigs exhibited significantly reduced human IgG and IgM binding. These results indicate that the antigenicity of TKO pig is significantly reduced and the remaining xenoantigens on porcine tissues can be eliminated via a gene targeting approach.

Keywords

pig / xenotransplantation / GGTA1 / CMAH / β4GalNT2 / antigenicity

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Ronggen Wang, Miaomiao Ruan, Runjie Zhang, Lei Chen, Xiaoxue Li, Bin Fang, Chu Li, Xueyang Ren, Jiying Liu, Qiang Xiong, Lining Zhang, Yong Jin, Lin Li, Rongfeng Li, Ying Wang, Haiyuan Yang, Yifan Dai. Antigenicity of tissues and organs from GGTA1/CMAH/b4GalNT2 triple gene knockout pigs. Journal of Biomedical Research, https://doi.org/10.7555/JBR.32.20180018

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Acknowledgments

This work was supported by grants from the National Natural Science Foundation of China (81570402 & 31701283), the National Key R&D Program of China (2017YFC1103701 & 2017YFC1103702), the Jiangsu Key Laboratory of Xenotransplantation (BM2012116), the Sanming Project of Medicine in Shenzhen, the Fund for High Level Medical Discipline Construction of Shenzhen (2016031638), and the Shenzhen Foundation of Science and Technology (JCYJ20160229204849975 & GCZX2015043017281705).

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2018 2018 by the Journal of Biomedical Research.
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