Epidermal growth factor receptor is a host-entry cofactor triggering hepatitis B virus internalization

Abstract

Sodium taurocholate cotransporting polypeptide (NTCP) is a host cell receptor required for hepatitis B virus (HBV) entry. However, the susceptibility of NTCP-expressing cells to HBV is diverse depending on the culture condition. Stimulation with epidermal growth factor (EGF) was found to potentiate cell susceptibility to HBV infection. Here, we show that EGF receptor (EGFR) plays a critical role in HBV virion internalization. In EGFR-knockdown cells, HBV or its preS1-specific fluorescence peptide attached to the cell surface, but its internalization was attenuated. PreS1 internalization and HBV infection could be rescued by complementation with functional EGFR. Interestingly, the HBV/preS1–NTCP complex at the cell surface was internalized concomitant with the endocytotic relocalization of EGFR. Molecular interaction between NTCP and EGFR was documented by immunoprecipitation assay. Upon dissociation from functional EGFR, NTCP no longer functioned to support viral infection, as demonstrated by either (i) the introduction of NTCP point mutation that disrupted its interaction with EGFR, (ii) the detrimental effect of decoy peptide interrupting the NTCP–EGFR interaction, or (iii) the pharmacological inactivation of EGFR. Together, these data support the crucial role of EGFR in mediating HBV–NTCP internalization into susceptible cells. EGFR thus provides a yet unidentified missing link from the cell-surface HBV–NTCP attachment to the viral invasion beyond the host cell membrane.

Host receptors: the key to establishing cells with broad viral tropism for vaccine production

“…This hypothesis, once tested true, will aid in our efforts towards the establishment of novel cell system for cost-efficient vaccine production that is adaptable to new viral strains or emerging viruses. However, we are aware that viruses presenting totally different types of genome (classified into different Baltimore subtypes) may share the same organ tropism and entry factors such as in the case of HBV and HCV that both target liver and use EGFR (Lupberger et al 2011;Iwamoto et al 2019) and NTCP (Yan et al 2012;Verrier et al 2016) for viral entry, and those belonging to the same Baltimore group may use different receptors for cell entry such as the differential use of HVEM, Nectin 1/2, GFR, CD63, CD151 in mediating the entry of type I viruses besides integrin (Table 1).…”

Can we cure hepatitis B virus with novel direct‐acting antivirals?

“…The HBV life cycle presents multiple potential targets for antiviral therapies. HBV infection is dependent upon the presence of its cellular receptor sodium taurocholate cotransporting polypeptide (NTCP) and recent data suggest that it is enhanced by the presence of epidermal growth factor (EGFR) . Once HBV is internalized, it releases its nucleocapsid to the cytoplasm, which travels to the hepatocyte nucleus to release the relaxed circular DNA (rcDNA).…”

“…HBV infection is dependent upon the presence of its cellular receptor sodium taurocholate cotransporting polypeptide (NTCP) 1 and recent data suggest that it is enhanced by the presence of epidermal growth factor (EGFR). 2 Once HBV is internalized, it releases its nucleocapsid to the cytoplasm, which travels to the hepatocyte nucleus to release the relaxed circular DNA (rcDNA). cccDNA originates from the incoming rcDNA by mechanisms that are not fully understood and associates with histone and non-histone proteins to build a minichromosome that acts as the only transcriptional template for all viral RNA.…”

Defined host factors support HBV infection in non‐hepatic 293T cells

“…NTCP physically interacts with the N‐terminal domain of preS1 and mediates viral entry into hepatocytes. Most recently, epidermal growth factor receptor has been discovered to play a critical role in mediating HBV‐NTCP internalization into hepatocytes . With the identification of the receptor, hepatoma cell lines that overexpress NTCP are susceptible to HBV infection .…”

“…Most recently, epidermal growth factor receptor has been discovered to play a critical role in mediating HBV-NTCP internalization into hepatocytes. 23 With the identification of the receptor, hepatoma cell lines that overexpress NTCP are susceptible to HBV infection. 14,24 As a bile salt transporter, NTCP is strongly expressed in parenchymal liver cells and is localized to the sinusoidal membrane.…”