Stem cell-derived hepatocytes: A novel model for hepatitis E virus replication

详细信息    查看全文

• 作者：Nicky Helsen¹ ; ^&dagger ; ; ^{nicky.helsen@med.kuleuven.be" class="auth_mail" title="E-mail the corresponding author} ; Yannick Debing² ; ^&dagger ; ; Jan Paeshuyse² ; Kai Dallmeier² ; Ruben Boon¹ ; Mar Coll³ ; Pau Sancho-Bru³ ; Christel Claes¹ ; Johan Neyts² ; ^&Dagger ; ; Catherine M. Verfaillie¹ ; ^&Dagger ;
• 关键词：(+)ssRNA ; positive strand RNA ; (&minus ; )ssRNA ; negative strand RNA ; αSMA ; alpha smooth muscle actin ; AAT ; alpha 1-antitrypsin ; AFP ; alpha fetoprotein ; BLBP ; brain lipid-binding protein ; BMP4 ; bone morphogenetic protein 4 ; COL1A1 ; collagen type 1 alpha 1 ; DLX2 ; distal-less homeobox 2 ; DMEM ; Dulbecco&rsquo ; s modified Eagle&rsquo ; s medium ; FBS ; fetal bovine serum ; FGF ; fibroblast growth factor ; FISH ; fluorescence in situ hybridization ; GAPDH ; glyceraldehyde-3-phosphate dehydrogenase ; hESC ; human em
• 刊名：Journal of Hepatology
• 出版年：2016
• 出版时间：March 2016
• 年：2016
• 卷：64
• 期：3
• 页码：565-573
• 全文大小：1466 K

文摘

Yearly, approximately 20 million people become infected with the hepatitis E virus (HEV) resulting in over 3 million cases of acute hepatitis. Although HEV-mediated hepatitis is usually self-limiting, severe cases of fulminant hepatitis as well as chronic infections have been reported, resulting annually in an estimated 60,000 deaths. We studied whether pluripotent stem cell (PSC)-derived hepatocytes, mesodermal and/or neuroprogenitor cells support HEV replication.

Methods

Human PSC were differentiated towards hepatocyte-like cells, mesodermal cells and neuroprogenitors and subsequently infected with HEV. Infection and replication of HEV was analyzed by qRT-PCR, RNA in situ hybridization, negative strand RT-PCR, production of infectious virions and transfection with a transient HEV reporter replicon.

Results

PSC-derived hepatocytes supported the complete replication cycle of HEV, as demonstrated by the intracellular presence of positive and negative strand HEV RNA and the production of infectious virions. The replication of the virus in these cells was inhibited by the antiviral drugs ribavirin and interferon-α2b. In contrast to PSC-derived hepatocytes, PSC-derived mesodermal cells and neuroprogenitors only supported HEV replication upon transfection with a HEV subgenomic replicon.

Conclusion

We demonstrate that PSC can be used to study the hepatotropism of HEV infection. The complete replication cycle of HEV can be recapitulated in infected PSC-derived hepatocytes. By contrast other germ layer cells support intracellular replication but are not infectable with HEV. Thus the early steps in the viral cycle are the main determinant governing HEV tissue tropism. PSC-hepatocytes offer a physiological relevant tool to study the biology of HEV infection and replication and may aid in the design of therapeutic strategies.