Yunnan Province Key Laboratory for Porcine Gene Editing and Xenotransplantation, Yunnan Agricultural University, Kunming, China.
Yunnan Province Xenotransplantation Research Engineering Centre, Yunnan Agricultural University, Kunming, China.
Front Immunol. 2022 Aug 9;13:950194. doi: 10.3389/fimmu.2022.950194. eCollection 2022.
Human hepatocyte transplantation for liver disease treatment have been hampered by the lack of quality human hepatocytes. Pigs with their large body size, longevity and physiological similarities with human are appropriate animal models for the expansion of human hepatocytes. Here we report on the generation of RAG2IL2RγFAH (RGFKO) pigs CRISPR/Cas9 system and somatic cell nuclear transfer. We showed that thymic and splenic development in RGFKO pigs was impaired. V(D)J recombination processes were also inactivated. Consequently, RGFKO pigs had significantly reduced numbers of porcine T, B and NK cells. Moreover, due to the loss of FAH, porcine hepatocytes continuously undergo apoptosis and consequently suffer hepatic damage. Thus, RGFKO pigs are both immune deficient and constantly suffer liver injury in the absence of NTBC supplementation. These results suggest that RGFKO pigs have the potential to be engrafted with human hepatocytes without immune rejection, thereby allowing for large scale expansion of human hepatocytes.
用于治疗肝脏疾病的人源肝细胞移植受到缺乏高质量人源肝细胞的阻碍。猪体型较大，与人类的生理相似，是扩增人源肝细胞的合适动物模型。在这里，我们报告了 RAG2IL2RγFAH (RGFKO) 猪的生成、CRISPR/Cas9 系统和体细胞核移植。我们表明，RGFKO 猪的胸腺和脾脏发育受损。V(D)J 重组过程也被失活。因此，RGFKO 猪的猪 T、B 和 NK 细胞数量明显减少。此外，由于 FAH 的缺失，猪肝细胞不断发生细胞凋亡，进而导致肝损伤。因此，在没有 NTBC 补充的情况下，RGFKO 猪既免疫缺陷又持续遭受肝损伤。这些结果表明，RGFKO 猪有可能在没有免疫排斥的情况下移植人源肝细胞，从而允许大规模扩增人源肝细胞。
