Fumarylacetoacetate Hydrolase Knock-out Rabbit Model for Hereditary Tyrosinemia Type 1.
Fumarylacetoacetate Hydrolase Knock-out Rabbit Model for Hereditary Tyrosinemia Type 1.
复制标题
富马酰乙酰乙酸水解酶敲除兔遗传性酪氨酸血症1型模型
DOI：
10.1074/jbc.m116.764787
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发表时间：
2017-03-17
期刊：
影响因子：
--
通讯作者：
Lai L
中科院分区：
文献类型：
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作者：
Li L;Zhang Q;Yang H;Zou Q;Lai C;Jiang F;Zhao P;Luo Z;Yang J;Chen Q;Wang Y;Newsome PN;Frampton J;Maxwell PH;Li W;Chen S;Wang D;Siu TS;Tam S;Tse HF;Qin B;Bao X;Esteban MA;Lai L
遗传性酪氨酸血症1型（HT 1）是一种严重的人类常染色体隐性遗传疾病，由富马酰乙酰乙酸羟化酶（FAH）缺乏引起，FAH是催化酪氨酸降解途径中最后一步的酶。缺乏FAH会导致血液和组织中有毒代谢物（富马酰乙酰乙酸酯和琥珀酰丙酮）的积累，最终导致严重的肝脏和肾脏损伤，从婴儿期到青春期发病。这种组织损伤是致命的，但可以通过给予2-（2-硝基-4-三氟甲基苯甲酰基）-1，3-环己二酮（NTBC）来控制，NTBC抑制富马酰乙酰乙酸酯和琥珀酰丙酮生成上游的酪氨酸催化剂。值得注意的是，在缺乏FAH的动物中，NTBC的短暂撤回可用于诱导肝损伤和刺激健康肝细胞生长的伴随再生反应。除此之外，该模型引起了人们对这些动物体内人原代肝细胞扩增以及探索实验性基因治疗和基于细胞的治疗的极大兴趣。在这里，我们报告一代FAH基因敲除兔通过原核阶段胚胎显微注射转录激活因子样效应核酸酶。FAH−/−兔表现出HT 1的表型特征，包括肝脏和肾脏异常，但还出现频繁的眼部表现，可能是NTBC给药后酪氨酸局部蓄积所致。我们还表明，将野生型兔原代肝细胞同种异体移植到FAH−/−兔中可以实现高效的肝脏再生，并防止肝功能不全和死亡。由于与啮齿动物相比具有显着的优势，并且与包括猪在内的大型动物相比易于繁殖，维护和操作，FAH−/−兔是模拟HT 1后果的有吸引力的替代品。
Hereditary tyrosinemia type 1 (HT1) is a severe human autosomal recessive disorder caused by the deficiency of fumarylacetoacetate hydroxylase (FAH), an enzyme catalyzing the last step in the tyrosine degradation pathway. Lack of FAH causes accumulation of toxic metabolites (fumarylacetoacetate and succinylacetone) in blood and tissues, ultimately resulting in severe liver and kidney damage with onset that ranges from infancy to adolescence. This tissue damage is lethal but can be controlled by administration of 2-(2-nitro-4-trifluoromethylbenzoyl)-1,3-cyclohexanedione (NTBC), which inhibits tyrosine catabolism upstream of the generation of fumarylacetoacetate and succinylacetone. Notably, in animals lacking FAH, transient withdrawal of NTBC can be used to induce liver damage and a concomitant regenerative response that stimulates the growth of healthy hepatocytes. Among other things, this model has raised tremendous interest for the in vivo expansion of human primary hepatocytes inside these animals and for exploring experimental gene therapy and cell-based therapies. Here, we report the generation of FAH knock-out rabbits via pronuclear stage embryo microinjection of transcription activator-like effector nucleases. FAH−/− rabbits exhibit phenotypic features of HT1 including liver and kidney abnormalities but additionally develop frequent ocular manifestations likely caused by local accumulation of tyrosine upon NTBC administration. We also show that allogeneic transplantation of wild-type rabbit primary hepatocytes into FAH−/− rabbits enables highly efficient liver repopulation and prevents liver insufficiency and death. Because of significant advantages over rodents and their ease of breeding, maintenance, and manipulation compared with larger animals including pigs, FAH−/− rabbits are an attractive alternative for modeling the consequences of HT1.