On the Host Side of the Hepatitis E Virus Life Cycle
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cells Review On the Host Side of the Hepatitis E Virus Life Cycle Noémie Oechslin , Darius Moradpour and Jérôme Gouttenoire * Division of Gastroenterology and Hepatology, Lausanne University Hospital and University of Lausanne, CH-1011 Lausanne, Switzerland; [email protected] (N.O.); [email protected] (D.M.) * Correspondence: [email protected] Received: 28 April 2020; Accepted: 21 May 2020; Published: 22 May 2020 Abstract: Hepatitis E virus (HEV) infection is one of the most common causes of acute hepatitis in the world. HEV is an enterically transmitted positive-strand RNA virus found as a non-enveloped particle in bile as well as stool and as a quasi-enveloped particle in blood. Current understanding of the molecular mechanisms and host factors involved in productive HEV infection is incomplete, but recently developed model systems have facilitated rapid progress in this area. Here, we provide an overview of the HEV life cycle with a focus on the host factors required for viral entry, RNA replication, assembly and release. Further developments of HEV model systems and novel technologies should yield a broader picture in the future. Keywords: HEV; host factor; particle production; viral replication; virus entry 1. Introduction Hepatitis E virus (HEV) has been identified as a cause of the waterborne hepatitis outbreaks in the early 1980s [1,2]. The viral genome was cloned and sequenced in 1990, allowing the development of serological tests to study its epidemiology [3,4]. The virus has been classified in the Hepeviridae family, and most human pathogenic strains belong to species Orthohepevirus A [5]. Members of this species can be classified into 8 genotypes (gt): gt 1 and 2 are restricted to humans and are transmitted via the fecal-oral route, mainly through contaminated drinking water. Gt 3 and 4 cause zoonotic infections and the transmission occurs mainly via the consumption of un(der)cooked pork, wild boar or deer meat. Gt 5 and 6 are found in wild boar, and gt 7 as well as 8 infect dromedary and Bactrian camels, respectively. Gt 7 has been identified in an immunosuppressed patient after consumption of camel milk and meat [6] (reviewed in [7]) but no transmission to humans has thus far been reported for gt 5, 6 and 8. More recently, rabbit HEV (closely related to gt 3 within species Orthohepevirus A) and rat HEV (belonging to species Orthohepevirus C) have also been found to infect humans [8–11]. HEV is a small, non-enveloped, icosahedral virus with a diameter of 27–34 nm [2]. It contains a 7.2 kb single-stranded, positive-sense RNA genome which possesses a m7G cap at its 50 and a poly-A tail at its 30 end (Figure1A). The HEV genome harbors 3 open reading frames (ORF). ORF1 encodes the viral replicase, ORF2 the capsid and ORF3 a small protein involved in virion secretion via its potential ion channel activity [12]. The first contact between HEV and host cells occurs through interaction with as yet poorly characterized entry factor(s). After endocytosis, the viral genome is released into the cytoplasm and the host translational machinery produces the ORF1 replicase, which drives viral RNA replication (Figure1B). During this step, two RNA species are produced from a negative-strand RNA intermediate: a full-length genomic RNA and a subgenomic RNA of 2.2 kb [13,14]. Translation of the subgenomic RNA yields the ORF2 and ORF3 proteins. Later steps of the HEV life cycle include viral assembly and release of newly produced virions. Very similar to hepatitis A virus (HAV), another hepatotropic positive-strand RNA virus, HEV is found as a ‘quasi-enveloped’ virion (eHEV) wrapped in exosomal membranes in blood and as a naked particle in bile and feces (reviewed in [15]) (Figure1B). Cells 2020, 9, 1294; doi:10.3390/cells9051294 www.mdpi.com/journal/cells Cells 2020, 9, 1294 2 of 14 Cells 2020, 9, x FOR PEER REVIEW 2 of 14 FigureFigure 1.1. Genome organization andand life cycle of hepatitis E virus (HEV).(HEV). (AA)) The 7.2 kb positive-strandpositive-strand 77 RNARNA genomegenome hashas aa 550′ 7-methylguanylate7-methylguanylate capcap (m(m G cap)cap) andand aa 330′ polyadenylatedpolyadenylated tailtail (poly-A).(poly-A). ItIt harborsharbors 33 open open reading reading frames frames (ORFs). (ORFs). ORF1 ORF1 encodes encodes a replicase a replicase of about of 190about kDa 190 comprising kDa comprising different functionaldifferent functional domains, including domains, a including methyltransferase a methyltransferase (Met), an RNA (Met), helicase an (Hel)RNA and helicase an RNA-dependent (Hel) and an RNARNA- polymerasedependent (RdRp),RNA polymerase as well as (RdRp) less well-characterized, as well as less well domains,-characterized such as the domains Y domain,, such a putativeas the Y papain-likedomain, a putative cysteine papain protease-like (PCP), cysteine a hypervariableprotease (PCP), region a hypervariable (HVR) and region the Macro (HVR) domain. and the M ORF2acro anddomain. ORF3 ORF2 encode and the ORF3 viral capsidencode andthe aviral small capsid protein and involved a small in protein virus secretion involved respectively, in virus secretion which arerespectively, translated whic fromh aare 2.2 translated kb subgenomic from a RNA2.2 kb generatedsubgenomic during RNA viralgenerated replication. during ( Bviral) The replication. HEV life cycle(B) The can HEV be dissected life cycle intocan be the dissected following into steps: the following (1) viral entry steps: by (1) as viral yet unidentifiedentry by as yet receptor(s), unidentified (2) endocytosisreceptor(s), (2) and endocytosis release of the and viral release positive-strand of the viral positive RNA genome-strand ( +RNA) into genome the cytosol, (+) into (3) the translation cytosol, of(3) the translation ORF1 protein of the to ORF1 allow protein replication to allow of the replication full-length of and the generation full-length of and the subgenomicgeneration of RNA the throughsubgenomic a negative-strand RNA through RNAa negative intermediate-strand RNA (-), (4) intermediate translation of(-), the (4) subgenomictranslation of RNA the subgenomic to produce theRNA ORF2 to produce and ORF3 the proteinsORF2 and and ORF3 (5) genome proteins packaging, and (5) genome virion packaging, assembly andvirion release assembly of the and virus release into theof the bloodstream virus into the and bloodstream the bile from and the the basolateral bile from andthe basolateral apical sides, and respectively. apical sides, ER, respectively. endoplasmic ER, reticulum;endoplasmic MVB, reticulum; multivesicular MVB, multivesicular body. body. As obligate intracellular pathogens, viruses have developed strategies to hijack and manipulate host cell pathways in order to ensure productive infection. Moreover, RNA viruses, especially those with relatively limited genome size and coding capacity, such as HEV, are particularly dependent on Cells 2020, 9, 1294 3 of 14 As obligate intracellular pathogens, viruses have developed strategies to hijack and manipulate host cell pathways in order to ensure productive infection. Moreover, RNA viruses, especially those with relatively limited genome size and coding capacity, such as HEV, are particularly dependent on the host cell machinery. Because the tools to study HEV have been limited until recently, only little is known about the host factors involved in the various steps of the viral life cycle. Studies performed in heterologous settings, such as the yeast two hybrid system, identified cellular factors interacting with HEVCells 20 proteins20, 9, x FOR [16 PEER,17]. REVIEW However, most of these candidates remain to be validated and further studied4 of 14 using infectious cell culture systems, in vivo models and liver biopsies from patients with hepatitis late endosomal and lysosomal Niemann-Pick disease type C1 protein, involved in cholesterol E. In this review, we shall focus on host factors whose involvement in the viral life cycle has been extraction, significantly reduced eHEV infection [22]. Moreover, treatment with an inhibitor of validated in HEV infection settings. lysosomal acid lipase, responsible for lipid degradation, resulted in a dose-dependent reduction of 2.eHEV HEV cell Entry entry [22]. These observations suggest that the quasi-envelope of eHEV is removed in endolysosomes. The viral Ascapsid HEV may is present subsequently in a non-enveloped interact with ("naked") an as yet and unidentified a quasi-enveloped host factor form (eHEV),and undergo the entry the pathwayconformational of the changes virus may required differ for for geno theseme tworelease forms. into the Our cytoplasm. knowledge A similar of HEV mechanism entry remains was recentlyscarce but proposed studies usingfor HAV virus-like [31]. particlesSince both (VLP) quasi as- aenveloped model system and havenon-enveloped highlighted HEV possible are internalizedhost factors in involved vesicles inbelonging the initial to the attachment endosomal to pathway, the cell it and is plausible virus internalization, that they use a including common β hostthe 78-kDa factor glucose-regulatedto allow uncoating protein and release (GRP78), of the ATP genome synthase into subunit the cytoplasm(ATPB5) (detailed and asialoglycoprotein in [43]). receptorThe (ASGPR)ORF3 protein [18–20 is]. present Notably, within non-enveloped eHEV and HEVinteracts was with shown the to capsid interact [44] with. However, heparan its sulfate role proteoglycansin eHEV entry, (HSPG),uncoating likely and genome syndecans release [21,22 remains], which to arebe explored. expressed