Genetically edited hepatic cells expressing the NTCP-S267F variant are resistant to hepatitis B virus infection.
Genetically edited hepatic cells expressing the NTCP-S267F variant are resistant to hepatitis B virus infection.
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表达NTCP-S267F变异体的基因编辑的肝细胞对乙肝病毒感染具有抵抗力。
DOI：
10.1016/j.omtm.2021.11.002
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发表时间：
2021-12-10
期刊：
影响因子：
--
通讯作者：
Liang TJ
中科院分区：
文献类型：
--
作者：
Uchida T;Park SB;Inuzuka T;Zhang M;Allen JN;Chayama K;Liang TJ
已知钠依赖性牛磺胆酸共转运多肽 (NTCP)-S267F 变体与乙型肝炎病毒 (HBV) 感染和疾病进展风险降低相关。 NTCP-S267F 变体在介导 HBV 进入方面表现出减弱的功能，但其在 HBV 感染中的功能尚未在更多生物学相关模型中完全确立。我们引入了 NTCP-S267F 变体，并在基因编辑的肝细胞中测试了 HBV 的感染性。经过 CRISPR 碱基编辑后，鉴定出具有纯合和杂合变体的 HepG2-NTCP 克隆。 NTCP-S267F 纯合克隆不支持 HBV 感染。杂合子克隆的行为与野生型克隆相似。我们生成了带有 NTCP-S267F 变体的基因编辑人类干细胞，其与野生型一样分化为表达高水平肝细胞分化标记物的肝细胞样细胞 (HLC)。我们证实，具有纯合变异的 HLC 不支持 HBV 感染，而杂合变异克隆与野生型细胞一样，同样会被 HBV 感染。总之，我们成功地将S267F变异体通过CRISPR碱基编辑导入到肝细胞的NTCP/SLC10A基因中，并证明该变异体是一种功能丧失突变。这种在自然环境下研究遗传变异及其发病机制的技术对于乙型肝炎的治疗干预具有潜在价值。作者利用 CRISPR 碱基编辑将 S267F 多态性引入人肝细胞的 HBV 受体 NTCP 基因中。他们描述了纯合或杂合 NTCP-S267F 变体对 HBV 感染的功能影响，并表明该变体是一种功能丧失突变。这些发现对乙型肝炎的发病机制和治疗具有重要意义。
The sodium-dependent taurocholate co-transporting polypeptide (NTCP)-S267F variant is known to be associated with a reduced risk of hepatitis B virus (HBV) infection and disease progression. The NTCP-S267F variant displays diminished function in mediating HBV entry, but its function in HBV infection has not been fully established in more biologically relevant models. We introduced the NTCP-S267F variant and tested infectivity by HBV in genetically edited hepatic cells. HepG2-NTCP clones with both homozygous and heterozygous variants were identified after CRISPR base editing. NTCP-S267F homozygous clones did not support HBV infection. The heterozygote clones behaved similarly to wild-type clones. We generated genetically edited human stem cells with the NTCP-S267F variant, which differentiated equally well as wild-type into hepatocyte-like cells (HLCs) expressing high levels of hepatocyte differentiation markers. We confirmed that HLCs with homozygous variant did not support HBV infection, and heterozygous variant clones were infected with HBV equally as well as the wild-type cells. In conclusion, we successfully introduced the S267F variant by CRISPR base editing into the NTCP/SLC10A gene of hepatocytes, and showed that the variant is a loss-of-function mutation. This technology of studying genetic variants and their pathogenesis in a natural context is potentially valuable for therapeutic intervention against HBV. The authors introduced the S267F polymorphism into the HBV receptor NTCP gene of human hepatic cells using CRISPR base editing. They characterized the functional effect of homozygous or heterozygous NTCP-S267F variant on HBV infection and showed that the variant is a loss-of-function mutation. The findings have implications in the pathogenesis and therapy of HBV.
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病毒进入：回顾并前进。
影响因子：
5.6
作者：
Helenius A
通讯作者：
Helenius A
通过表达优化和祖先重建改进胞苷和腺嘌呤碱基编辑器。
影响因子：
46.9
作者：
Koblan LW;Doman JL;Wilson C;Levy JM;Tay T;Newby GA;Maianti JP;Raguram A;Liu DR
通讯作者：
Liu DR
DOI：
10.1002/hep.28025
发表时间：
2015-12
期刊：
Hepatology (Baltimore, Md.)
影响因子：
--
作者：
Liang TJ;Block TM;McMahon BJ;Ghany MG;Urban S;Guo JT;Locarnini S;Zoulim F;Chang KM;Lok AS
通讯作者：
Lok AS
影响因子：
25.7
作者：
Li W;Urban S
通讯作者：
Urban S
影响因子：
6
作者：
Ishida, Yuji;Yamasaki, Chihiro;Tateno, Chise
通讯作者：
Tateno, Chise