Genetically edited hepatic cells expressing the NTCP-S267F variant are resistant to hepatitis B virus infection

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...

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Autores principales:	Takuro Uchida, Seung Bum Park, Tadashi Inuzuka, Min Zhang, Joselyn N. Allen, Kazuaki Chayama, T. Jake Liang
Formato:	article
Lenguaje:	EN
Publicado:	Elsevier 2021
Materias:	HBV NTCP S267F polymorphism CRISPR base editing stem cell-derived hepatocyte Genetics QH426-470 Cytology QH573-671
Acceso en línea:	https://doaj.org/article/6da94754d7634c86bd34d99931061100
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spelling	oai:doaj.org-article:6da94754d7634c86bd34d999310611002021-11-22T04:26:40ZGenetically edited hepatic cells expressing the NTCP-S267F variant are resistant to hepatitis B virus infection2329-050110.1016/j.omtm.2021.11.002https://doaj.org/article/6da94754d7634c86bd34d999310611002021-12-01T00:00:00Zhttp://www.sciencedirect.com/science/article/pii/S2329050121001741https://doaj.org/toc/2329-0501The 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.Takuro UchidaSeung Bum ParkTadashi InuzukaMin ZhangJoselyn N. AllenKazuaki ChayamaT. Jake LiangElsevierarticleHBVNTCPS267F polymorphismCRISPR base editingstem cell-derived hepatocyteGeneticsQH426-470CytologyQH573-671ENMolecular Therapy: Methods & Clinical Development, Vol 23, Iss , Pp 597-605 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	HBV NTCP S267F polymorphism CRISPR base editing stem cell-derived hepatocyte Genetics QH426-470 Cytology QH573-671
spellingShingle	HBV NTCP S267F polymorphism CRISPR base editing stem cell-derived hepatocyte Genetics QH426-470 Cytology QH573-671 Takuro Uchida Seung Bum Park Tadashi Inuzuka Min Zhang Joselyn N. Allen Kazuaki Chayama T. Jake Liang Genetically edited hepatic cells expressing the NTCP-S267F variant are resistant to hepatitis B virus infection
description	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.
format	article
author	Takuro Uchida Seung Bum Park Tadashi Inuzuka Min Zhang Joselyn N. Allen Kazuaki Chayama T. Jake Liang
author_facet	Takuro Uchida Seung Bum Park Tadashi Inuzuka Min Zhang Joselyn N. Allen Kazuaki Chayama T. Jake Liang
author_sort	Takuro Uchida
title	Genetically edited hepatic cells expressing the NTCP-S267F variant are resistant to hepatitis B virus infection
title_short	Genetically edited hepatic cells expressing the NTCP-S267F variant are resistant to hepatitis B virus infection
title_full	Genetically edited hepatic cells expressing the NTCP-S267F variant are resistant to hepatitis B virus infection
title_fullStr	Genetically edited hepatic cells expressing the NTCP-S267F variant are resistant to hepatitis B virus infection
title_full_unstemmed	Genetically edited hepatic cells expressing the NTCP-S267F variant are resistant to hepatitis B virus infection
title_sort	genetically edited hepatic cells expressing the ntcp-s267f variant are resistant to hepatitis b virus infection
publisher	Elsevier
publishDate	2021
url	https://doaj.org/article/6da94754d7634c86bd34d99931061100
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Genetically edited hepatic cells expressing the NTCP-S267F variant are resistant to hepatitis B virus infection

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