Modifications of Adenovirus Hexon Allow for Either Hepatocyte Detargeting or Targeting With Potential Evasion From Kupffer Cells

Prill, Jan-Michael; Espenlaub, Sigrid; Samen, Ulrike; Engler, Tatjana; Schmidt, Erika; Vetrini, Francesco; Rosewell, Amanda; Grove, Nathan; Palmer, D. Jason; Ng, Philip; Kochanek, Stefan; Kreppel, Florian

doi:10.1038/mt.2010.229

Cited by 68 publications

(57 citation statements)

References 37 publications

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“…Therefore, we predicted that the PEGylated cysteinemodified viruses should retain in vitro activity unless a critical HVR was involved in virus entry events. Previous data targeting PEGylation to HVR5 of Ad5 has shown that viral activity is retained (27), supporting the concept that targeted modification of all seven HVRs may similarly retain in vitro activity.…”

Section: Resultsmentioning

confidence: 62%

“…To probe HVR-specific interactions with SRA-II and Kupffer cells, we applied targeted PEGylation to all seven of the Ad5 HVRs using the approach that previously was used to target PEG to HVR5 (11,27). By this systematic approach, targeted PEGylation of HVRs 1, 2, 5, and 7 produced marked increases in liver hepatocyte transduction in vivo after intravenous injection.…”

Section: Discussionmentioning

confidence: 99%

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Identification of Adenovirus Serotype 5 Hexon Regions That Interact with Scavenger Receptors

Khare

Reddy

Nemerow

et al. 2012

J Virol

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Most of an intravenous dose of species C adenovirus serotype 5 (Ad5) is destroyed by liver Kupffer cells. In contrast, another species C virus, Ad6, evades these cells to mediate more efficient liver gene delivery. Given that this difference in Kupffer cell interaction is mediated by the hypervariable (HVR) loops of the virus hexon protein, we genetically modified each of the seven HVRs of Ad5 with a cysteine residue to enable conditional blocking of these sites with polyethylene glycol (PEG). We show that these modifications do not affect in vitro virus transduction. In contrast, after intravenous injection, targeted PEGylation at HVRs 1, 2, 5, and 7 increased viral liver transduction up to 20-fold. Elimination or saturation of liver Kupffer cells did not significantly affect this increase in the liver transduction. In vitro, PEGylation blocked uptake of viruses via the Kupffer cell scavenger receptor SRA-II. These data suggest that HVRs 1, 2, 5, and 7 of Ad5 may be involved in Kupffer cell recognition and subsequent destruction. These data also demonstrate that this conditional genetic-chemical mutation strategy is a useful tool for investigating the interactions of viruses with host tissues.A denovirus serotype 5 (Ad5) has proven to be one of the most potent in vivo gene delivery vectors for liver-directed gene therapy. As much as 95 to 98% of an intravenously (i.v.) injected dose of Ad5 is trafficked to the liver (12). The adenovirus capsid is comprised of three major proteins: hexon, penton base, and fiber (reviewed in reference 7). Based on numerous in vitro studies, adenovirus fiber and penton base have been shown to be cellular attachment proteins. The fiber of Ad5 binds coxsackie and adenovirus receptor (CAR) (3), which triggers binding of penton base to ␣v integrins via an RGD motif (35,36) and results in viral cell internalization. Although necessary for providing specificity of the virus in vitro, modifications to the fiber have little effect on vector tropism in vivo (25). Instead, increasing evidence suggests that hexon plays a large role in the natural liver tropism of Ad5.Hexon is the most abundant viral capsid protein with 720 monomers per virion. Hexon organizes into trimers so that three hexon monomers and their loops wrap tightly around each other to create a tower-like structure with a central depression (20,28). Each hexon monomer has seven flexible, serotype-specific loops, named hypervariable regions (HVRs) (23), that are predicted to be located on the surface of the hexon trimer and the virion (29). This location allows the HVRs of hexon to interact with neutralizing antibodies, receptors, proteins, and cells. Considering there are 5,040 (720 ϫ 7 ϭ 5,040) HVRs per virion, these represent a complex, exposed surface for many interactions.After i.v. injection, Ad5 exhibits the greatest transduction within liver hepatocytes (12). Despite this robust in vivo gene delivery, ϳ90% of the injected dose is sequestered and destroyed by resident liver macrophages called Kupffer cells (1). These anti...

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Section: Resultsmentioning

confidence: 62%

Section: Discussionmentioning

confidence: 99%

Identification of Adenovirus Serotype 5 Hexon Regions That Interact with Scavenger Receptors

Khare

Reddy

Nemerow

et al. 2012

J Virol

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“…Pronounced tumoral iodide uptake even after 15-18 d demonstrates high transduction efficiency and efficient replication of Ad5-E1/AFP-E3/NIS in the tumor. Evasion of the hybrid vector from scavenging by Kupffer cells could be the possible mechanism behind the observed liver detargeting of transgene expression, as it was suggested by the study of Prill et al (34).…”

Section: Discussionmentioning

confidence: 59%

Systemic Image-Guided Liver Cancer Radiovirotherapy Using Dendrimer-Coated Adenovirus Encoding the Sodium Iodide Symporter as Theranostic Gene

et al. 2013

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Currently, major limitations for the clinical application of adenovirus-mediated gene therapy are high prevalence of neutralizing antibodies, widespread expression of the coxsackie-adenovirus receptor (CAR), and adenovirus sequestration by the liver. In the current study, we used the sodium iodide symporter (NIS) as a theranostic gene to investigate whether coating of adenovirus with synthetic dendrimers could be useful to overcome these hurdles in order to develop adenoviral vectors for combination of systemic oncolytic virotherapy and NIS-mediated radiotherapy. Methods: We coated replication-deficient (Ad5-CMV/NIS) (CMV is cytomegalovirus) and replication-selective (Ad5-E1/AFP-E3/NIS) adenovirus serotype 5 carrying the hNIS gene with poly(amidoamine) dendrimers generation 5 (PAMAM-G5) in order to investigate transduction efficacy and altered tropism of these coated virus particles by 123 I scintigraphy and to evaluate their therapeutic potential for systemic radiovirotherapy in a liver cancer xenograft mouse model. Results: After dendrimer coating, Ad5-CMV/NIS demonstrated partial protection from neutralizing antibodies and enhanced transduction efficacy in CAR-negative cells in vitro. In vivo 123 I scintigraphy of nude mice revealed significantly reduced levels of hepatic transgene expression after intravenous injection of dendrimer-coated Ad5-CMV/NIS (dcAd5-CMV/NIS). Evasion from liver accumulation resulted in significantly reduced liver toxicity and increased transduction efficiency of dcAd5-CMV/NIS in hepatoma xenografts. After PAMAM-G5 coating of the replication-selective Ad5-E1/AFP-E3/NIS, a significantly enhanced oncolytic effect was observed after intravenous application (virotherapy) that was further increased by additional treatment with a therapeutic dose of 131 I (radiovirotherapy) and was associated with markedly improved survival. Conclusion: These results demonstrate efficient liver detargeting and tumor retargeting of adenoviral vectors after coating with synthetic dendrimers, thereby representing a promising innovative strategy for systemic NIS gene therapy. Moreover, our study-based on the function of NIS as a theranostic gene allowing the noninvasive imaging of NIS expression by 123 I scintigraphy-provides detailed characterization of in vivo vector biodistribution and localization, level, and duration of transgene expression, essential prerequisites for exact planning and monitoring of clinical gene therapy trials that aim to individualize the NIS gene therapy concept.

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“…However, modification on vector tropism was not clearly reported in this study (Park et al, 2010). Although a great part of systemically administrated vector particles is scavenged by Kupffer cells in the liver, the use of a PEG-or a dextran-coated vectors allowed the specific transduction of hepatocytes independent of the presence of Kupffer cells, emphasizing the potential for therapeutic liver-directed gene transfer (Prill et al, 2010). Similar result can be observed by using a multivalent hydrophilic polymer based on poly-[N-(2-hydroxypropyl)methacrylamide] conjugated with EGF or VCAM, producing a CARindependent binding and uptake into EGF-or VCAM-positive target cells selectively in mixed culture and also in xenografts in-vivo (Fisher et al, 2001).…”

Section: Chemical Modification Of the Adv Capsidmentioning

confidence: 95%

Gene Delivery Systems: Tailoring Vectors to Reach Specific Tissues

Alméciga-Díaz¹,

Cuaspa²,

Barrera³

2011

Non-Viral Gene Therapy

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Modifications of Adenovirus Hexon Allow for Either Hepatocyte Detargeting or Targeting With Potential Evasion From Kupffer Cells

Cited by 68 publications

References 37 publications

Identification of Adenovirus Serotype 5 Hexon Regions That Interact with Scavenger Receptors

Identification of Adenovirus Serotype 5 Hexon Regions That Interact with Scavenger Receptors

Systemic Image-Guided Liver Cancer Radiovirotherapy Using Dendrimer-Coated Adenovirus Encoding the Sodium Iodide Symporter as Theranostic Gene

Gene Delivery Systems: Tailoring Vectors to Reach Specific Tissues

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