An interaction between penton base and αv integrins plays a minimal role in adenovirus-mediated gene transfer to hepatocytes in vitro and in vivo

Hautala, Timo; Grunst, Teresa; Fabrega, Alfredo J.; Freimuth, Paul; Welsh, M. J.

doi:10.1038/sj.gt.3300722

Cited by 22 publications

(16 citation statements)

References 19 publications

(38 reference statements)

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“…The absence of significant accumulation of radioactivity in the liver suggests that once bound to CAR molecules on the hepatocyte surface, knob is rapidly delivered to an intracellular compartment where it is exposed to deiodinase activity. Consistent with this model of integrin-independent uptake of knob, an earlier study showed that adenovirus-CAR complexes on rat hepatocytes can be internalized by an integrin-independent pathway (18).…”

Section: Discussionsupporting

confidence: 60%

Biodistribution of Radioiodinated Adenovirus Fiber Protein Knob Domain after Intravenous Injection in Mice

Awasthi

Meinken

Springer

et al. 2004

J Virol

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The knob domains from the fiber proteins of adenovirus serotypes 2 and 12 were labeled with radioiodine and then injected into the bloodstreams of mice. Knob proteins with functional binding sites for the coxsackie and adenovirus receptor (CAR) were cleared rapidly from the circulation, with radioactivity appearing predominantly in the stomach, while knob mutants unable to bind to CAR remained in the blood circulation for a prolonged period. The clearance of radiolabeled wild-type knob from the blood was slowed by coinjecting an excess of unlabeled wild-type knob protein. An earlier study showed that 99m Tc-labeled knob protein with intact CAR-binding activity also cleared rapidly from the blood circulation of mice, with radioactivity accumulating predominantly in the liver (K. R. Zinn et al., Gene Ther. 5:798-808, 1998). Together these results suggest that rapid clearance of knob protein from the blood results from specific binding to CAR in the liver and that the bound knob then enters a degradative pathway. The elevated levels of radioiodine in the stomach observed in our experiments are consistent with deiodination of labeled knob by dehalogenases in hepatocyte microsomes and uptake of the resultant free radioiodine by Na/I symporters in the gastric mucosa. Although CAR has been shown to localize in tight junctions of polarized epithelial cells, where it functions in intercellular adhesion, the results of our study suggest that a subset of CAR molecules in the liver is highly accessible to ligands in the blood and able to rapidly deliver bound ligand to an intracellular degradative compartment.

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Section: Discussionsupporting

confidence: 60%

Biodistribution of Radioiodinated Adenovirus Fiber Protein Knob Domain after Intravenous Injection in Mice

Awasthi

Meinken

Springer

et al. 2004

J Virol

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“…However, MHC class I level was not observed to be associated with adenovirus transduction efficiency, 34,37 though it was found to be adenovirus type 5 fiber receptor. 15 Furthermore, integrins v 3 and v 5 did not appear to be absolutely necessary for efficient adenovirus transduction of respiratory epithelial cells, 38 39 in contrast to previous reports that integrins v 3 and v 5 were necessary for adenovirus entry. 16,24 Nevertheless, it was suggested that integrins v 3 and v 5 might be facilitative rather than necessary or limiting for efficient adenovirus entry into the host cell.…”

Section: Discussionmentioning

confidence: 74%

Coxsackievirus–adenovirus receptor expression in ovarian cancer cell lines is associated with increased adenovirus transduction efficiency and transgene expression

et al. 2001

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The expression of coxsackievirus ± adenovirus receptor ( CAR ) and the integrins v 3 and v 5 was analyzed quantitatively ( flow cytometry ) and qualitatively ( immunocytochemistry ) in five human ovarian cancer cell lines ( PEO1, PEO4, PEO14, SKOV -3, and OVCAR -3 ) and three control cell lines ( 293, HeLa, and CHO -K1 ) . The transduction efficiencies were evaluated by adv / rsv --Gal transduction followed by X -gal staining. The effects of 17 -estradiol on cell growth, CAR and integrins v 3 / 5 expression, adenovirus transduction efficiency, and cell -killing efficacy of adv / rsv -tk plus ganciclovir were determined. The levels of CAR, integrin v 3, and integrin v 5 showed great variation between the cell lines. Whereas the expression of CAR appeared to be essential for and positively correlated with adenovirus transduction efficiency, the integrins v 3 and v 5 were not absolutely necessary for adenovirus transduction even though their presence may facilitate transduction. In PEO4 and PEO1 cells, proliferation was stimulated by 17 -estradiol in a dose -dependent manner. In PEO4 cells, and much less pronounced in PEO1 cells, this was accompanied by an increase in CAR expression. The stimulation of CAR expression was paralleled by an increased transduction efficiency resulting in an increased cell -killing efficacy. Our data suggest that the expression of CAR is one of the most important prerequisites for successful adenovirus -mediated gene therapy of ovarian cancer. Cancer Gene Therapy ( 2001 ) 8, 168 ± 175

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“…For example, hepatocytes express the fiber receptor, CAR, but not the penton receptors, a v integrins, yet are infected efficiently by Ad2. 33 Similarly, canine adenovirus type 2 (CAV-2) and Ad40/41 both efficiently bind CAR but enter cells through an RGD-independent pathway. [34][35][36] In our ongoing studies, we are investigating the molecular and cellular requirements for fiber uptake.…”

Section: Discussionmentioning

confidence: 99%

The Ad5 fiber mediates nonviral gene transfer in the absence of the whole virus, utilizing a novel cell entry pathway

et al. 2004

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The interesting discovery reported here that soluble adenovirus serotype 5 (Ad5) fiber proteins enter cells without the virus was a serendipitous result during our development of Ad5 capsid proteins as nonviral gene transfer vectors. The Ad5 capsid fiber and penton proteins mediate infection. The fiber docks to a noninternalizing cell surface protein called the coxsackievirus-Ad receptor (CAR), followed by penton binding to integrins, triggering integrin-mediated endocytosis of the virus. In our previous work, we assembled the nonviral complex, 3PO, which utilized the penton to mediate gene transfer through integrin binding and endocytosis. Here, we tested whether incorporating the fiber targets 3PO to CAR, thus recapitulating the Ad5 infection pathway. As CAR is not an endocytic receptor, we were surprised to find that the fiber alone, without the penton, enabled gene transfer by binding CAR, but internalizing through an unknown mechanism. We show here that the fiber distributes to the nucleus and cytoplasm after temperature-independent uptake, whereas the penton accumulates around the nucleus after temperature-dependent uptake. Fiber uptake by HeLa cells is also actin-dependent, requires the fiber tail/shaft region, and is largely inhibited by heparin. This study raises the possibility that alternative pathways may enable both viral and nonviral cell entry. Gene Therapy (2005) 12, 225-237.

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An interaction between penton base and αv integrins plays a minimal role in adenovirus-mediated gene transfer to hepatocytes in vitro and in vivo

Cited by 22 publications

References 19 publications

Biodistribution of Radioiodinated Adenovirus Fiber Protein Knob Domain after Intravenous Injection in Mice

Biodistribution of Radioiodinated Adenovirus Fiber Protein Knob Domain after Intravenous Injection in Mice

Coxsackievirus–adenovirus receptor expression in ovarian cancer cell lines is associated with increased adenovirus transduction efficiency and transgene expression

The Ad5 fiber mediates nonviral gene transfer in the absence of the whole virus, utilizing a novel cell entry pathway

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