Capsid-mediated control of adeno-associated viral transcription determines host range.

Adeno-associated virus (AAV) is a member of the genus Dependoparvovirus, which infects a wide range of vertebrate species. Here, we observe that, unlike most primate AAV isolates, avian AAV is transcriptionally silenced in human cells. By swapping the VP1 N terminus from primate AAVs (e.g., AAV8) onto non-mammalian isolates (e.g., avian AAV), we identify a minimal component of the AAV capsid that controls viral transcription and unlocks robust transduction in both human cells and mouse tissue. This effect is accompanied by increased AAV genome chromatin accessibility and altered histone methylation. Proximity ligation analysis reveals that host factors are selectively recruited by the VP1 N terminus of AAV8 but not avian AAV. Notably, these include AAV essential factors implicated in the nuclear factor κB pathway, chromatin condensation, and histone methylation. We postulate that the AAV capsid has evolved mechanisms to recruit host factors to its genome, allowing transcriptional activation in a species-specific manner.

A Novel Dependoparvovirus Identified in Cloacal Swabs of Monk Parakeet (Myiopsitta monachus) from Urban Areas of Spain.

The introduction of invasive birds into new ecosystems frequently has negative consequences for the resident populations. Accordingly, the increasing population of monk parakeets (Myiopsitta monachus) in Europe may pose a threat because we have little knowledge of the viruses they can transmit to native naïve species. In this study, we describe a new dependoparvovirus detected by metagenomic analysis of cloacal samples from 28 apparently healthy individuals captured in urban areas of Madrid, Spain. The genomic characterization revealed that the genome encoded the NS and VP proteins typical of parvoviruses and was flanked by inverted terminal repeats. No recombination signal was detected. The phylogenetic analysis showed that it was closely related to a parvovirus isolated in a wild psittacid in China. Both viruses share 80% Rep protein sequence identity and only 64% with other dependoparvoviruses identified in Passeriformes, Anseriformes, and Piciformes and are included in a highly supported clade, which could be considered a new species. The prevalence was very low, and none of the additional 73 individuals tested positive by PCR. These results highlight the importance of exploring the viral genome in invasive species to prevent the emergence of novel viral pathogenic species.

Characterizing two novel goose parvoviruses with different origins.

Outbreaks of short beak dwarf syndrome caused by novel goose parvovirus (NGPV) have been prevalent in China since 2015, resulting in a high mortality rate of ducks. Herein we evaluated differences between two NGPV strains: Muscovy duck-origin (AH190917-RP: MD17) and Cherry Valley duck-origin (JS191021-RP: CVD21) NGPV. Both of them showed certain level of pathogenicity to primary duck embryo fibroblasts, Cherry Valley duck embryos and ducklings. CVD21 showed comparatively stronger pathogenicity than MD17. Only CVD21 caused obvious cytopathic effect (CPE), characterized by cell shedding; further, the virus titer of MD17 and CVD21 was 102.571 ELD50 (i.e. median embryo lethal dose)/0.2 ml and 106.156 ELD50 /0.2 ml, respectively, and the mortality rate of CVD21- and MD17-infected Cherry Valley ducklings was 100% and 80%, respectively. In addition, CVD21 had a greater influence on the growth and development of ducklings. Furthermore, we found that MD17 could infect Muscovy duck embryos and produce lesions similar to Cherry Valley duck embryos, but it could not infect Muscovy duck embryo fibroblasts (MDEFs,) and Muscovy ducklings. MDV21 had no infection to MDEFs, Muscovy duck embryo and Muscovy ducklings. We then sequenced the complete genome of the two isolates to enable genomic characterization. The complete genome of MD17 and CVD21 was 5046 and 5050 nucleotides in length, respectively. Nucleotide alignment, amino acid analysis and phylogenetic tree analysis revealed that MD17 showed higher homology to goose parvovirus (GPV), while CVD21 demonstrated stronger similarity with NGPV. Moreover, the two isolates shared 95.8% homology, with encoded proteins showing multiple amino acid variations. Our findings indicate that Muscovy ducks seem to have played a crucial role in the evolution of GPV to NGPV. We believe that our data should serve as a foundation for further studying the genetic evolution of waterfowl parvoviruses and their pathogenic mechanisms.

Evolution of dependoparvoviruses across geological timescales-implications for design of AAV-based gene therapy vectors.

Endogenous viral elements (EVEs) are genetic remnants of viruses that have integrated into host genomes millions of years ago and retained as heritable elements passed on to offspring until present-day. As a result, EVEs provide an opportunity to analyse the genomes of extinct viruses utilizing these genomic viral fossils to study evolution of viruses over large timescales. Analysis of sequences from near full-length EVEs of dependoparvoviral origin identified within three mammalian taxa, Whippomorpha (whales and hippos), Vespertilionidae (smooth-nosed bats), and Lagomorpha (rabbits, hares, and pikas), indicates that distinct ancestral dependoparvovirus species integrated into these host genomes approximately 77 to 23 million years ago. These ancestral viruses are unique relative to modern adeno-associated viruses (AAVs), and distinct from extant species of genus Dependoparvovirus. These EVE sequences show characteristics previously unseen in modern, mammalian AAVs, but instead appear more similar to the more primitive, autonomously replicating and pathogenic waterfowl dependoparvoviruses. Phylogeny reconstruction suggests that the whippomorph EVE orthologue derives from exogenous ancestors of autonomous and highly pathogenic dependoparvovirus lineages, believed to have uniquely co-evolved with waterfowl birds to present date. In contrast, ancestors of the two other mammalian orthologues (Lagomorpha and Vespertilionidae) likely shared the same lineage as all other known mammalian exogenous AAVs. Comparative in silico analysis of the EVE genomes revealed remarkable overall conservation of AAV rep and cap genes, despite millions of years of integration within the host germline. Modelling these proteins identified unexpected variety, even between orthologues, in previously defined capsid viral protein (VP) variable regions, especially in those related to the three- and fivefold symmetry axes of the capsid. Moreover, the normally well-conserved phospholipase A2 domain of the predicted minor VP1 also exhibited a high degree of sequence variance. These findings may indicate unique biological properties for these virus 'fossils' relative to extant dependoparvoviruses and suggest key regions to explore within capsid sequences that may confer novel properties for engineered gene therapy vectors based on paleovirology data.

Parvovirus Family Conundrum: What Makes a Killer?

Parvoviruses infect a wide variety of hosts, and their ancestors appear to have emerged tens to hundreds of millions of years ago and to have spread widely ever since. The diversity of parvoviruses is therefore extensive, and although they all appear to descend from a common ancestor and share common structures in their capsid and nonstructural proteins, there is often low homology at the DNA or protein level. The diversity of these viruses is also seen in the widely differing impacts they have on their hosts, which range from severe and even lethal disease to subclinical or nonpathogenic infections. In the past few years, deep sequencing of DNA samples from animals has shown just how widespread the parvoviruses are in nature, but most of the newly discovered viruses have not yet been associated with any disease. However, variants of some parvoviruses have altered their host ranges to create new epidemic or pandemic viruses. Here, we examine the properties of parvoviruses and their interactions with their hosts that are associated with these disparate pathogenic outcomes.