Variant enterovirus A71 found in immune-suppressed patient binds to heparan sulfate and exhibits neurotropism in B-cell-depleted mice

Weng, Kuo-Feng; Tee, Han Kang; Tseligka, Eirini D.; Cagno, Valeria; Mathez, Gregory; Rosset, Stéphane; Nagamine, Claude M.; Sarnow, Peter; Kirkegaard, Karla; Tapparel, Caroline

doi:10.1016/j.celrep.2023.112389

Cited by 2 publications

(6 citation statements)

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“…As previously observed for strong HS-binding variants, we showed that, in contrast to the original mouse-adapted MP4 strain which exhibited virulence in mice, this cell-adapted MP4-97R/167G double mutant was completely attenuated in mice 19 . In the current study, we used MP4 and MP4-97R/167G mutant as representatives of respectively, weak and strong HS-binders, slow and fast-growing in cell lines and virulent and avirulent in mouse models (as documented previously 19,20 ), to elucidate the consequence of virus adaptation towards HS binding on the viral growth cycle. We demonstrated that these mutations not only increase binding to HS, but also reduce capsid stability, leading to improved uncoating and faster cell internalization in a HS-independent manner.…”

Section: Introductionsupporting

confidence: 85%

“…Of note, although strong HS-binder are clearly attenuated in mice, the situation in humans is more puzzling as strains with Q or G at position VP1-145 have been associated with severe neurological cases and the methodology used in those studies excluded emergence of mutation during cell culture 52,53 . Furthermore, we previously showed that intermediate HS-binding affinity can result in increased virulence even in mouse models 19 . It is still unclear if there is a lack of trapping and/or limited impact on viral stability under these conditions.…”

Section: Discussionmentioning

confidence: 97%

“…The situation may differ significantly in vivo as strong HS binders are attenuated. Koike and colleagues have demonstrated that strong binding to HS induces virus trapping in vivo 9-11,19 . Our data suggest that the associated decreased stability may further contribute to viral attenuation.…”

Section: Discussionmentioning

confidence: 99%

“…We previously isolated cell-adapted EV-A71 mutants with strong affinity for HS which emerged upon passaging of intermediate HS binders derived from both patient and mouse-adapted MP4 strains in cell culture 18,19 . The mutants presented two amino acid changes in the VP1 capsid protein: VP1-L97R mutation in the VP1 BC loop, shown to confer intermediate affinity for HS together with a secondary mutation, VP1-E167G, located in the VP1 EF loop, which significantly strengthened HS binding with reduction of negative charges 19,20 . As previously observed for strong HS-binding variants, we showed that, in contrast to the original mouse-adapted MP4 strain which exhibited virulence in mice, this cell-adapted MP4-97R/167G double mutant was completely attenuated in mice 19 .…”

Section: Introductionmentioning

confidence: 99%

“…The mutants presented two amino acid changes in the VP1 capsid protein: VP1-L97R mutation in the VP1 BC loop, shown to confer intermediate affinity for HS together with a secondary mutation, VP1-E167G, located in the VP1 EF loop, which significantly strengthened HS binding with reduction of negative charges 19,20 . As previously observed for strong HS-binding variants, we showed that, in contrast to the original mouse-adapted MP4 strain which exhibited virulence in mice, this cell-adapted MP4-97R/167G double mutant was completely attenuated in mice 19 . In the current study, we used MP4 and MP4-97R/167G mutant as representatives of respectively, weak and strong HS-binders, slow and fast-growing in cell lines and virulent and avirulent in mouse models (as documented previously 19,20 ), to elucidate the consequence of virus adaptation towards HS binding on the viral growth cycle.…”

Section: Introductionmentioning

confidence: 99%

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Enterovirus A71 adaptation to heparan sulfate comes with capsid stability tradeoff

Tee,

Mathez,

Cagno

et al. 2024

Preprint

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SummaryBecause of high mutation rates, viruses constantly adapt to new environments. When propagated in cell lines, certain viruses acquire positively charged amino acids on their surface proteins, enabling them to utilize negatively charged heparan sulfate (HS) as an attachment receptor. In this study, we used enterovirus A71 (EV-A71) as model and demonstrated that unlike the parental MP4 variant, the cell-adapted strong HS-binder MP4-97R/167G does not require acidification for uncoating and releases its genome in the neutral or weakly acidic environment of early endosomes. We experimentally confirmed that this pH-independent entry is not associated with the use of HS as an attachment receptor but rather with compromised capsid stability. We then extended these findings to another HS-dependent strain, suggesting that adaptation to HS generally modifies capsid stability and alters entry mechanism. Our data show EV-A71 pH-independent entry for the first time and, more importantly, highlight the intricate interplay between HS-binding, capsid stability, and viral fitness, wherein enhanced multiplication in cell lines leads to attenuation in hostilein vivoenvironments such as the gastrointestinal tract.

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Section: Introductionsupporting

confidence: 85%

Section: Discussionmentioning

confidence: 97%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Enterovirus A71 adaptation to heparan sulfate comes with capsid stability tradeoff

Tee,

Mathez,

Cagno

et al. 2024

Preprint

Self Cite

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Novel virulence determinants in VP1 regulate the assembly of enterovirus-A71

Zhang,

Li,

et al. 2024

J Virol

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Enterovirus-A71 (EV-A71) is the second most common causative agent after coxsackievirus A16 of hand, foot, and mouth disease. The capsids of EV-A71 consist of 60 copies of each of the four viral structural proteins (VP1–VP4). VP1 is highly exposed and surface accessible, playing a central role in virus particle assembly, attachment, and entry. To gain insight into the role of highly conserved residues at positions 75, 78, and 88 in the capsid protein VP1 in these processes, an alanine-scanning analysis was performed using an infectious cDNA clone of EV-A71. Our study revealed that the substitutions of VP1-T75A, VP1-T78A, and VP1-G88A could affect the assembly of the virus capsid proteins, resulting in the production of abnormal virions with reduced infectivity. Specifically, the substitution of VP1-T75A affected the maturation cleavage of the VP0 precursor, leading to deficiencies in binding to receptor scavenger receptor class B2 (SCARB2), viral attachment, internalization, and even uncoating. For the mutants of T78A and G88A, a significant reduction in virion-associated genomic RNA was observed, suggesting that more noninfectious empty particles were produced during viral assembly. Interestingly, the VP1-T75A variant showed weak replication in cell cultures but demonstrated increased virulence in BALB/c neonatal mice, which might be due to the difference in viral receptors among mammalian species. Taken together, our data revealed the important role of the highly conserved residues T75, T78, and G88 in VP1 protein in the infectivity of EV-A71. Characterizing these novel determinants of EV-A71 virulence would contribute to rationally developing effective treatments and broadly protective vaccine candidates. IMPORTANCE EV-A71 causes hand, foot, and mouth disease in children. In this study, we discovered three highly conserved residues at positions 75, 78, and 88 of the capsid protein VP1 as the potential virulence determinants of EV-A71, which can influence viral replication by regulating the assembly of EV-A71. Mechanistic studies revealed that VP1-T75A could affect the maturation cleavage of the VP0 precursor, resulting in deficiencies in binding to the receptor SCARB2, viral attachment, internalization, and even uncoating. For the mutants of T78A and G88A, more noninfectious empty particles were produced during viral assembly. The discovery of these novel determinants of EV-A71 virulence will promote the study of the pathogenesis of enteroviruses.

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Variant enterovirus A71 found in immune-suppressed patient binds to heparan sulfate and exhibits neurotropism in B-cell-depleted mice

Cited by 2 publications

References 53 publications

Enterovirus A71 adaptation to heparan sulfate comes with capsid stability tradeoff

Enterovirus A71 adaptation to heparan sulfate comes with capsid stability tradeoff

Novel virulence determinants in VP1 regulate the assembly of enterovirus-A71

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