Modulation of Sialic Acid Dependence Influences the Central Nervous System Transduction Profile of Adeno-associated Viruses

Albright, Blake H.; Simon, Katherine E.; Pillai, Minakshi; Devlin, Garth; Asokan, Aravind

doi:10.1128/jvi.00332-19

Cited by 55 publications

(10 citation statements)

References 43 publications

(55 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…6D). In a recent study, the amino acids of AAVrh.10 VR-I were substituted to the corresponding sequences of AAV1 (AAVRX1) and the BBB-crossing efficiency was reduced in this variant but not completely lost (64). This indicates that additional amino acids of the AAVrh.10 capsid act together with serine 269 to achieve the BBB crossing phenotype.…”

Section: Resultsmentioning

confidence: 99%

Comparative Analysis of the Capsid Structures of AAVrh.10, AAVrh.39, and AAV8

Mietzsch

Barnes

Hull

et al. 2020

J Virol

View full text Add to dashboard Cite

Adeno-associated viruses (AAVs) from clade E are often used as vectors in gene delivery applications. This clade includes rhesus isolate 10 (AAVrh.10) and 39 (AAVrh.39) which, unlike representative AAV8, are capable of crossing the blood-brain barrier (BBB), thereby enabling the delivery of therapeutic genes to the central nervous system. Here, the capsid structures of AAV8, AAVrh.10 and AAVrh.39 have been determined by cryo-electron microscopy and three-dimensional image reconstruction to 3.08-, 2.75-, and 3.39-Å resolution, respectively, to enable a direct structural comparison. AAVrh.10 and AAVrh.39 are 98% identical in amino acid sequence but only ∼93.5% identical to AAV8. However, the capsid structures of all three viruses are similar, with only minor differences observed in the previously described surface variable regions, suggesting that specific residues S269 and N472, absent in AAV8, may confer the ability to cross the BBB in AAVrh.10 and AAVrh.39. Head-to-head comparison of empty and genome-containing particles showed DNA ordered in the previously described nucleotide-binding pocket, supporting the suggested role of this pocket in DNA packaging for the Dependoparvovirus. The structural characterization of these viruses provides a platform for future vector engineering efforts toward improved gene delivery success with respect to specific tissue targeting, transduction efficiency, antigenicity, or receptor retargeting. IMPORTANCE Recombinant adeno-associated virus vectors (rAAVs), based on AAV8 and AAVrh.10, have been utilized in multiple clinical trials to treat different monogenetic diseases. The closely related AAVrh.39 has also shown promise in vivo. As recently attained for other AAV biologics, e.g., Luxturna and Zolgensma, based on AAV2 and AAV9, respectively, the vectors in this study will likely gain U.S. Food and Drug Administration approval for commercialization in the near future. This study characterized the capsid structures of these clinical vectors at atomic resolution using cryo-electron microscopy and image reconstruction for comparative analysis. The analysis suggested two key residues, S269 and N472, as determinants of BBB crossing for AAVrh.10 and AAVrh.39, a feature utilized for central nervous system delivery of therapeutic genes. The structure information thus provides a platform for engineering to improve receptor retargeting or tissue specificity. These are important challenges in the field that need attention. Capsid structure information also provides knowledge potentially applicable for regulatory product approval.

show abstract

Section: Resultsmentioning

confidence: 99%

Comparative Analysis of the Capsid Structures of AAVrh.10, AAVrh.39, and AAV8

Mietzsch

Barnes

Hull

et al. 2020

J Virol

View full text Add to dashboard Cite

show abstract

“…Additional observations that may test the predictions from insilico reports implicating a role for sialic acid residues as SARS-CoV-2 receptors include (1) efficacy of the therapeutic use of lactoferrin 66 , an antiviral agent that interacts with sialic acid residues; (2) an ongoing clinical trial of DAS181 (https:// clinicaltrials.gov/ct2/show/NCT04324489), a drug designed to block viral access by cleaving sialic acid; (3) that the shedding pattern of SARS-CoV-2 infection is different from that of SARS-CoV and more similar to that of "standard" influenza 66 , where sialic acid receptors play a major role; (4) a bioinformatic study reports binding of S-protein to sialic acid glycans in a region close to that identified by the in silico studies 65,67 . To our knowledge, while links between sialic acid and neurotropism for mouse hepatitis virus 68 and adenoviruses 69 have been suggested, there are no published investigations on this alternative pathway for SARS-CoV-2 interaction in the nervous system.…”

Section: Sars-cov-2 Receptors and Cellular Uptakementioning

confidence: 99%

COVID-19 and possible links with Parkinson’s disease and parkinsonism: from bench to bedside

Sulzer

Antonini

Leta

et al. 2020

npj Parkinsons Dis.

138

165

View full text Add to dashboard Cite

This Viewpoint discusses insights from basic science and clinical perspectives on coronavirus disease 2019 (COVID-19)/severe acute respiratory syndrome-coronavirus-2 (SARS-CoV-2) infection in the brain, with a particular focus on Parkinson's disease. Major points include that neuropathology studies have not answered the central issue of whether the virus enters central nervous system neurons, astrocytes or microglia, and the brain vascular cell types that express virus have not yet been identified. Currently, there is no clear evidence for human neuronal or astrocyte expression of angiotensin-converting enzyme 2 (ACE2), the major receptor for viral entry, but ACE2 expression may be activated by inflammation, and a comparison of healthy and infected brains is important. In contrast to the 1918 influenza pandemic and avian flu, reports of encephalopathy in COVID-19 have been slow to emerge, and there are so far no documented reports of parkinsonism apart from a single case report. We recommend consensus guidelines for the clinical treatment of Parkinson's patients with COVID-19. While a role for the virus in causing or exacerbating Parkinson's disease appears unlikely at this time, aggravation of specific motor and non-motor symptoms has been reported, and it will be important to monitor subjects after recovery, particularly for those with persisting hyposmia.

show abstract

“…Notably, AAV5 effectively crossed Caco-2 barrier epithelial monolayers; while competitive inhibition of AAV5 engagement with sialic acid reduced transduction, it did not ablate trancytosis, suggesting that AAV5 may utilize a distinct cellular entry pathway in crossing tissue layers (32). Previous work in our lab suggests that sialic acid binding affinity of AAV1 can be modulated by engrafting a minimal footprint of amino acid residues derived from AAVrh.10, which, in turn, can profoundly impact the ability to traverse the vascular endothelium/blood-brain barrier (38,39). Together, these data suggest that glycan receptor engagement and polarized receptor localization could play integral roles in determining AAV transduction versus transcellular transport profiles in vitro and vector biodistribution in vivo.…”

Section: Discussionmentioning

confidence: 96%

A CRISPR Screen Identifies the Cell Polarity Determinant Crumbs 3 as an Adeno-associated Virus Restriction Factor in Hepatocytes

Madigan

Tyson

Yuziuk

et al. 2019

J Virol

Self Cite

View full text Add to dashboard Cite

Adeno-associated viruses (AAV) are helper-dependent parvoviruses that have been developed into promising gene therapy vectors. Many studies, including a recent unbiased genomic screen, have identified host factors essential for AAV cell entry, but no genome-wide screens that address inhibitory host factors have been reported. Here, we utilize a novel CRISPR screen to identify AAV restriction factors in a human hepatocyte cell line. The major hit from our gain-of-function screen is the apical polarity determinant Crumbs 3 (Crb3). Knockout (KO) of Crb3 enhances AAV transduction, while overexpression exerts the opposite effect. Further, Crb3 appears to restrict AAV transduction in a serotype- and cell type-specific manner. Particularly, for AAV serotype 9 and a rationally engineered AAV variant, we demonstrate that increased availability of galactosylated glycans on the surfaces of Crb3 KO cells, but not the universal AAV receptor, leads to increased capsid attachment and enhanced transduction. We postulate that Crb3 could serve as a key molecular determinant that restricts the availability of AAV glycan attachment factors on the cell surface by maintaining apical-basal polarity and tight junction integrity.IMPORTANCEAdeno-associated viruses (AAVs) have recently emerged at the forefront as gene therapy vectors; however, our understanding of host factors that influence AAV transduction in different cell types is still evolving. In the present study, we perform a genome-scale CRISPR knockout screen to identify cellular host factors that restrict AAV infection in hepatocyte cultures. We discover that Crumbs 3, which determines cellular polarity, also influences the distribution of certain carbohydrate attachment factors on the cell surface. This in turn affects the ability of virions to bind and enter the cells. This study underscores the importance of cell polarity in AAV transduction and provides a potential molecular basis for the differential infectious mechanism(s) in cell culture versus organ systems.

show abstract

Modulation of Sialic Acid Dependence Influences the Central Nervous System Transduction Profile of Adeno-associated Viruses

Cited by 55 publications

References 43 publications

Comparative Analysis of the Capsid Structures of AAVrh.10, AAVrh.39, and AAV8

Comparative Analysis of the Capsid Structures of AAVrh.10, AAVrh.39, and AAV8

COVID-19 and possible links with Parkinson’s disease and parkinsonism: from bench to bedside

A CRISPR Screen Identifies the Cell Polarity Determinant Crumbs 3 as an Adeno-associated Virus Restriction Factor in Hepatocytes

Contact Info

Product

Resources

About