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Biology subjects

Viktorova, E. G.

Publications and source records attributed to Viktorova, E. G..

3 recordsLinked to original sources

Infection-specific long-chain fatty acid metabolism as a broad anti-enterovirus target

Enteroviruses are arguably the most numerous group of viruses infecting humans. While most enterovirus infections are benign and self-resolving, their sheer number inevitably increases the chances of multiple complications. The diversity of enteroviruses means that the development of vaccines is only economically feasible against a select few, and no direct-acting or host-targeted anti-virals are approved to treat enteroviral infections, largely due to the rapid development of resistance against all experimental drugs. Here, we explored a universal property of enterovirus infection - a massive upregulation of phospholipid synthesis as a target for anti-viral interventions. The increased phospholipid synthesis consumes endogenously- and exogenously-derived long-chain fatty acids (LCFA). We demonstrate that polyunsaturated LCFAs can have a broad anti-enteroviral effect, affecting multiple steps of the virus life cycle. The anti-viral activity of LCFAs did not strictly depend on the degree of unsaturation or their capacity to induce lipid peroxidation but significantly correlated with their conformation. This suggests that their incorporation into the phospholipid molecules makes the replication organelle membranes incapable of properly accommodating viral replication machinery. Accordingly, the inhibition of neutral lipid synthesis promoted LCFAs retargeting to the membranes in infected cells and increased their anti-viral potency. We show that this approach is effective against diverse enteroviruses in different cell types, including differentiated primary cells, and that attempts to establish viruses resistant to such treatment were unsuccessful.

microbiology↗

Viral vector-driven trans-encapsidation of replicon RNAs as a rapid approach for the development of safe and economically attractive anti-enterovirus vaccines

Multiple enteroviruses are associated with life-threatening and economically important diseases, yet licensed vaccines are available only against poliovirus (worldwide) and enterovirus A71 (China). Both live attenuated and inactivated anti-poliovirus vaccines, while highly successful in preventing the disease, have important shortcomings. Live vaccine strains are inherently genetically unstable and can regain virulence, leading to re-emergence of paralytic disease. Inactivated vaccine does not induce the mucosal immunity sufficient to interrupt viral transmission and is made from virulent strains, presenting a biosafety challenge. Recent alternatives, such as new vaccine strains with improved genetic stability and VLP-based vaccines, only partially address these concerns. Here, we investigated another approach to the development of anti-enterovirus vaccines based on efficient trans-encaspidation of replication-competent enterovirus RNAs coding for only the non-structural proteins (replicons) by Newcastle Disease virus vectors expressing enterovirus capsid proteins. Thus, the encapsidated replicon production is driven by effectively replicating enterovirus RNA and the NDV vector. This system is easily scalable and can be adapted to any cell culture provided it can be infected by both the enterovirus and NDV. Unlike the empty VLPs, the encapsidated replicons recapitulate the stability and antigenicity of native enterovirus particles, but cannot propagate beyond the originally infected cell. The protective efficacy of an encapsidated poliovirus replicon immunization was similar to that of the licensed Sabin vaccine strain in a murine model. This approach can easily be adapted to any enterovirus, allowing the rapid development of new, affordable vaccine candidates.

microbiology↗

The Development of Resistance to an Inhibitor of a Cellular Protein Reveals a critical interaction between the enterovirus protein 2C and a small GTPase Arf1

The cellular protein GBF1, an activator of Arf GTPases (ArfGEF: Arf guanine nucleotide exchange factor), is recruited to the replication organelles of enteroviruses through interaction with the viral protein 3A, and its ArfGEF activity is required for viral replication. Here, we investigated the development of resistance of poliovirus, a prototype enterovirus, to increasing concentrations of brefeldin A (BFA), an inhibitor of GBF1. High level of resistance required a gradual accumulation of multiple mutations in the viral protein 2C. The 2C mutations conferred BFA resistance even in the context of a 3A mutant previously shown to be defective in the recruitment of GBF1 to replication organelles, and in cells depleted of GBF1, suggesting a GBF1-independent replication mechanism. Still, activated Arfs accumulated on the replication organelles of this mutant even in the presence of BFA, its replication was inhibited by a pan-ArfGEF inhibitor LM11, and the BFA-resistant phenotype was compromised in Arf1-knockout cells. Importantly, the mutations strongly increased the interaction of 2C with the activated form of Arf1. Analysis of other enteroviruses revealed a particularly strong interaction of 2C of human rhinovirus 1A with activated Arf1. Accordingly, the replication of this virus was significantly less sensitive to BFA than that of poliovirus. Thus, our data demonstrate that enterovirus 2Cs may behave like Arf1 effector proteins and that GBF1 but not Arf activation can be dispensable for enterovirus replication. These findings have important implications for the development of host-targeted anti-viral therapeutics.

microbiology↗