Enhanced Gene Delivery to Human Primary Endothelial Cells Using Tropism-Modified Adenovirus Vectors

Preuss, Meredith A.; Glasgow, Joel N.; Everts, Maaike; Stoff-Khalili, Mariam A.; Wu, Hongju; Curiel, David T.

doi:10.2174/1875037000801010007

Cited by 13 publications

(9 citation statements)

References 28 publications

(23 reference statements)

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“…EC receptors targeted by this mechanism include VEGFR-2, Tie-2, E-selectin, endoglin, and membrane-bound angiotensin converting enzyme 7, 31–33 . EC transduction has also been achieved through capsid fiber knob display of peptide ligands such as the arginine-glycine-asparate (RGD) motif cognate for the angiogenesis associated integrins α v /β 5 and α v /β 3 29, 34 , or the DDTRHWG peptide 3 . A parallel strategy for EC specificity has been transcriptional targeting using enhancer/promoter elements of endothelial-specific genes such as VEGFR-2 , VEGFR-1 , preproendothelin-1 , and roundabout-4 10, 23, 35–38 .…”

Section: Discussionmentioning

confidence: 99%

The myeloid-binding peptide adenoviral vector enables multi-organ vascular endothelial gene targeting

Kaliberov

Zhang

et al. 2014

Laboratory Investigation

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Vascular endothelial cells (ECs) are ideal gene therapy targets as they provide widespread tissue access and are the first contact surfaces following intravenous vector administration. Human recombinant adenovirus serotype 5 (Ad5) is the most frequently used gene transfer system because of its appreciable transgene payload capacity and lack of somatic mutation risk. However, standard Ad5 vectors predominantly transduce liver but not the vasculature following intravenous administration. We recently developed an Ad5 vector with a myeloid cell-binding peptide (MBP) incorporated into the knob-deleted, T4 fibritin chimeric fiber (Ad.MBP)1.This vector was shown to transduce pulmonary ECs presumably via a vector handoff mechanism. Here we tested the body wide tropism of the Ad.MBP vector, its myeloid cell necessity, and vector-EC expression dose response. Using comprehensive multi-organ co-immunofluorescence analysis, we discovered that Ad.MBP produced widespread EC transduction in lung, heart, kidney, skeletal muscle, pancreas, small bowel, and brain. Surprisingly, Ad.MBP retained hepatocyte tropism albeit at a reduced frequency compared with standard Ad5. While binding specifically to myeloid cells ex vivo, multi-organ Ad.MBP expression was not dependent on circulating monocytes or macrophages. Ad.MBP dose de-escalation maintained full lung targeting capacity but drastically reduced transgene expression in other organs. Swapping the EC-specific ROBO4 for the CMV promoter/enhancer abrogated hepatocyte expression but also reduced gene expression in other organs. Collectively, our multilevel targeting strategy could enable therapeutic biologic production in previously inaccessible organs that initiate the most debilitating or lethal human diseases.

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Section: Discussionmentioning

confidence: 99%

The myeloid-binding peptide adenoviral vector enables multi-organ vascular endothelial gene targeting

Kaliberov

Zhang

et al. 2014

Laboratory Investigation

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“…The transductional targeting strategies seek to re-direct Ad5 binding to non-native receptors expressing on specific cell type. These approaches utilize Ad capsid display of peptides or bispecific antibodies cognate endothelial cell surface receptors (Glasgow, Everts, and Curiel, 2006; Kim et al, 2011; Preuss et al, 2008; Reynolds et al, 2001; Work et al, 2006) as well as Ad5 pseudotyping using fiber substitution with knob from different serotypes (Bachtarzi et al, 2011; Preuss et al, 2008; Shinozaki et al, 2006). Targeting of gene expression in the context of Ad employ can also be achieved with transcriptional targeting methods whereby transgenes are placed under the control of a tissue-specific promoter (Dong and Nor, 2009; Greenberger et al, 2004; Savontaus et al, 2002).…”

Section: Discussionmentioning

confidence: 99%

Retargeting of gene expression using endothelium specific hexon modified adenoviral vector

Kaliberov

Kaliberova

et al. 2013

Virology

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Adenovirus serotype 5 (Ad5) vectors are well suited for gene therapy. However, tissue-selective transduction by systemically administered Ad5-based vectors is confounded by viral particle sequestration in the liver. Hexon-modified Ad5 expressing reporter gene under transcriptional control by the immediate/early cytomegalovirus (CMV) or the Roundabout 4 receptor (Robo4) enhancer/promoter were characterized by growth in cell culture, stability in vitro, gene transfer in the presence of human coagulation factor X, and biodistribution in mice. The obtained data demonstrate the utility of the Robo4 promoter in an Ad5 vector context. Substitution of the hypervariable region 7 (HVR7) of the Ad5 hexon with HVR7 from Ad serotype 3 resulted in decreased liver tropism and dramatically altered biodistribution of gene expression. The results of these studies suggest that the combination of liver detargeting using a genetic modification of hexon with an endothelium-specific transcriptional control element produces an additive effect in the improvement of Ad5 biodistribution.

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“…Similarly, a comparable 6-fold reduction in spleen uptake and transduction was accompanied by decreased colocalisation of the β-galactosidase transgene with MAdCAM-1 in spleen sections from macrophage-depleted AdT*RGE animals compared to AdT* counterparts. Previous studies indicate that activation of sinusoidal endothelial cells in the liver after iv delivery of Ad5 is significantly attenuated by deletion of the penton base RGD motif [37,38], whilst in vitro transduction of human pulmonary artery, coronary artery and umbilical vein endothelial cells can be significantly increased by insertion of an RGD motif into the Ad5 fibre knob domain [39]. Taken together, these data suggest that uptake of Ad5 vectors by sinus-lining endothelial cells in the spleen may be mediated by an interaction between the penton base RGD motif and cellular integrins.…”

Section: Discussionmentioning

confidence: 99%

Biodistribution and inflammatory profiles of novel penton and hexon double-mutant serotype 5 adenoviruses

Bradshaw

Coughlan

Miller

et al. 2012

Journal of Controlled Release

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The use of adenovirus serotype 5 (Ad5) vectors in the clinical setting is severely hampered by the profound liver tropism observed after intravascular delivery coupled with the pronounced inflammatory and innate immune response elicited by these vectors. Liver transduction by circulating Ad5 virions is mediated by a high-affinity interaction between the capsid hexon protein and blood coagulation factor X (FX), whilst penton–αvintegrin interactions are thought to contribute to the induction of anti-Ad5 inflammatory and innate immune responses. To overcome these limitations, we sought to develop and characterise for the first time novel Ad5 vectors possessing mutations ablating both hexon:FX and penton:integrin interactions. As expected, intravascular administration of the FX binding-ablated Ad5HVR5*HVR7*E451Q vector (AdT*) resulted in significantly reduced liver transduction in vivo compared to Ad5. In macrophage-depleted mice, increased spleen uptake of AdT* was accompanied by an elevation in the levels of several inflammatory mediators. However ablation of the penton RGD motif in the AdT* vector background (AdT*RGE) resulted in a significant 5-fold reduction in spleen uptake and attenuated the antiviral inflammatory response. A reduction in spleen uptake and inflammatory activation was also observed in animals after intravascular administration of Ad5RGE compared to the parental Ad5 vector, with reduced co-localisation of the viral beta-galactosidase transgene with MAdCAM-1 + sinus-lining endothelial cells. Our detailed assessment of these novel adenoviruses indicates that penton base RGE mutation in combination with FX binding-ablation may be a viable strategy to attenuate the undesired liver uptake and pro-inflammatory responses to Ad5 vectors after intravascular delivery.

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Enhanced Gene Delivery to Human Primary Endothelial Cells Using Tropism-Modified Adenovirus Vectors

Cited by 13 publications

References 28 publications

The myeloid-binding peptide adenoviral vector enables multi-organ vascular endothelial gene targeting

The myeloid-binding peptide adenoviral vector enables multi-organ vascular endothelial gene targeting

Retargeting of gene expression using endothelium specific hexon modified adenoviral vector

Biodistribution and inflammatory profiles of novel penton and hexon double-mutant serotype 5 adenoviruses

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