Survival Advantage of Both Human Hepatocyte Xenografts and Genome-Edited Hepatocytes for Treatment of α-1 Antitrypsin Deficiency.
Survival Advantage of Both Human Hepatocyte Xenografts and Genome-Edited Hepatocytes for Treatment of α-1 Antitrypsin Deficiency.
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DOI：
10.1016/j.ymthe.2017.09.020
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发表时间：
2017-11-01
期刊：
影响因子：
--
通讯作者：
Mueller C
中科院分区：
文献类型：
--
作者：
Borel F;Tang Q;Gernoux G;Greer C;Wang Z;Barzel A;Kay MA;Shultz LD;Greiner DL;Flotte TR;Brehm MA;Mueller C
肝细胞是基因治疗和单基因疾病编辑的重要靶点。在α-1抗胰蛋白酶（AAT）缺乏症中，一个错义突变导致AAT分泌受损。在大多数患者中，由于缺乏AAT介导的肺弹性蛋白对中性粒细胞弹性蛋白酶的保护而发生肺损伤。在一些患者中，错误折叠的PiZ突变体AAT蛋白的积累触发肝细胞损伤，导致炎症和肝硬化。我们假设，纠正肝细胞中的Z突变缺陷将赋予完整肝脏内肝细胞再增殖的选择性优势。将人PiZ等位基因与免疫缺陷（NSG）株杂交以产生用于人肝细胞异种移植的受体株（NSG-PiZ）。结果表明，与NSG受体相比，NSG-PiZ受体支持正常人原代肝细胞的植入增加。因此，该模型可用于测试AATD的肝细胞疗法，但更广泛地说，它可作为一种简单、高度可重复的肝脏异种移植模型。最后，将表达野生型AAT和合成miRNA以沉默内源等位基因的无启动子腺相关病毒（AAV）载体整合到白蛋白基因座中。这种基因编辑方法通过沉默突变蛋白和增加正常AAT产生以及改善肝脏病理学，导致编辑肝细胞的选择性优势。Borel等人描述了两项基于错误折叠的人α-1抗胰蛋白酶（A1 AT）的研究。首先，当A1 AT在NSG小鼠的肝脏中表达时，它允许人肝细胞的可重复移植。第二，肝细胞的基因编辑以减少错误折叠的蛋白质，导致校正细胞的扩增和肝脏疾病的改善。
Hepatocytes represent an important target for gene therapy and editing of single-gene disorders. In α-1 antitrypsin (AAT) deficiency, one missense mutation results in impaired secretion of AAT. In most patients, lung damage occurs due to a lack of AAT-mediated protection of lung elastin from neutrophil elastase. In some patients, accumulation of misfolded PiZ mutant AAT protein triggers hepatocyte injury, leading to inflammation and cirrhosis. We hypothesized that correcting the Z mutant defect in hepatocytes would confer a selective advantage for repopulation of hepatocytes within an intact liver. A human PiZ allele was crossed onto an immune-deficient (NSG) strain to create a recipient strain (NSG-PiZ) for human hepatocyte xenotransplantation. Results indicate that NSG-PiZ recipients support heightened engraftment of normal human primary hepatocytes as compared with NSG recipients. This model can therefore be used to test hepatocyte cell therapies for AATD, but more broadly it serves as a simple, highly reproducible liver xenograft model. Finally, a promoterless adeno-associated virus (AAV) vector, expressing a wild-type AAT and a synthetic miRNA to silence the endogenous allele, was integrated into the albumin locus. This gene-editing approach leads to a selective advantage of edited hepatocytes, by silencing the mutant protein and augmenting normal AAT production, and improvement of the liver pathology. Borel et al. describe two studies based on misfolded human α-1 antitrypsin (A1AT). First, when A1AT is expressed in livers of NSG, mice it allows for reproducible engraftment of human hepatocytes. Second, gene editing of hepatocytes to decrease misfolded protein results in expansion of corrected cells and amelioration of liver disease.
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影响因子：
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DOI：
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期刊：
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