Elimination of GGTA1,CMAH,β4GalNT2 and CIITA genes in pigs compromises human versus pig xenogeneic immune reactions

Jing Xu ¹Jilong Ren ²Kai Xu ²Minghui Fang ³Meina Ka ¹Fei Xu ³Xin Wang ¹Jing Wang ¹Zhiqiang Han ¹Guihai Feng ²Ying Zhang ²Tang Hai ⁴Wei Li ⁵Zheng Hu³

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作者信息

1. State Key Laboratory of Stem Cell and Reproductive Biology,Institute of Zoology,Chinese Academy of Sciences,Beijing,China;Key Laboratory of Organ Regeneration and Reconstruction,Chinese Academy of Sciences,Beijing,China;University of Chinese Academy of Sciences,Beijing,China
2. State Key Laboratory of Stem Cell and Reproductive Biology,Institute of Zoology,Chinese Academy of Sciences,Beijing,China;Key Laboratory of Organ Regeneration and Reconstruction,Chinese Academy of Sciences,Beijing,China;Beijing Institute for Stem Cell and Regenerative Medicine,Beijing,China
3. Key Laboratory of Organ Regeneration and Transplantation of the Ministry of Education,The First Hospital,Jilin University,Changchun,China
4. State Key Laboratory of Stem Cell and Reproductive Biology,Institute of Zoology,Chinese Academy of Sciences,Beijing,China;Key Laboratory of Organ Regeneration and Reconstruction,Chinese Academy of Sciences,Beijing,China;Beijing Farm Animal Research Center,Institute of Zoology,Chinese Academy of Sciences,Beijing,China
5. State Key Laboratory of Stem Cell and Reproductive Biology,Institute of Zoology,Chinese Academy of Sciences,Beijing,China;Key Laboratory of Organ Regeneration and Reconstruction,Chinese Academy of Sciences,Beijing,China;Beijing Institute for Stem Cell and Regenerative Medicine,Beijing,China;University of Chinese Academy of Sciences,Beijing,China
折叠

Abstract

Background:Pig organ xenotransplantation is a potential solution for the severe organ shortage in clinic,while immunogenic genes need to be eliminated to im-prove the immune compatibility between humans and pigs.Current knockout strat-egies are mainly aimed at the genes causing hyperacute immune rejection (HAR) that occurs in the first few hours while adaptive immune reactions orchestrated by CD4 T cell thereafter also cause graft failure,in which process the MHC Ⅱ molecule plays critical roles.Methods:Thus,we generate a 4-gene ( GGTA1,CMAH,β4GalNT2,and CIITA ) knockout pig by CRISPR/Cas9 and somatic cell nuclear transfer to compromise HAR and CD4 T cell reactions simultaneously.Results:We successfully obtained 4KO piglets with deficiency in all alleles of genes,and at cellular and tissue levels.Additionally,the safety of our animals after gene editing was verified by using whole-genome sequencing and karyotyping.Piglets have survived for more than one year in the barrier,and also survived for more than 3 months in the conventional environment,suggesting that the piglets without MHC Ⅱ can be raised in the barrier and then gradually mated in the conventional environment.Conclusions:4KO piglets have lower immunogenicity,are safe in genomic level,and are easier to breed than the model with both MHC Ⅰ and Ⅱ deletion.

Key words

CD4 T cell/genetically edited pig/immune rejection/major histocompatibility complex Ⅱ/xenotransplantation

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基金项目

National Key Research and Development Program(2019YFA0903800)

National Key Research and Development Program(2021YFA0805701)

National Key Research and Development Program(2021YFA0805905)

National Key Research and Development Program(2022YFA1103603)

CAS Project for Young Scientists in Basic Research(YSBR-012)

STI 2030-Major Project(2023ZD0407503)

National Natural Science Foundation of China(32071456)

National Natural Science Foundation of China(82241224)

Strategic Priority Research Program of the Chinese Academy of Sciences(XDA16030000)

出版年

2024

动物模型与实验医学(英文)

中国实验动物学会,中国医学科学院医学实验动物研究所

动物模型与实验医学(英文)

ISSN：2096-5451

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