Adenovirus targeted to heparan-containing receptors increases its gene delivery efficiency to multiple cell types

Wickham, Thomas J.; Roelvink, Peter W.; Brough, Douglas E.; Kovesdi, Imre

doi:10.1038/nbt1196-1570

Cited by 301 publications

(229 citation statements)

References 18 publications

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“…Other investigators have genetically modified the adenovirus to include new binding motivs into the viral capsid 11 or the HI loop. 12 In these studies, the native binding to the CAR receptor was not blocked, and therefore vector tropism was not changed; additional tropism was added in these instances.…”

Section: Figure 3 Selective Infection Of Human Colorectal Cancer Cellmentioning

confidence: 99%

Tumor-specific gene transfer via an adenoviral vector targeted to the pan-carcinoma antigen EpCAM

et al. 1999

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Section: Figure 3 Selective Infection Of Human Colorectal Cancer Cellmentioning

confidence: 99%

Tumor-specific gene transfer via an adenoviral vector targeted to the pan-carcinoma antigen EpCAM

et al. 1999

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“…However, there is growing evidence that the expression of CAR is frequently suppressed in various types of cells, including tumor cells and in primary tumors [4,5], resulting in resistance to adenovirus infection [6][7][8]. Nevertheless, various studies have shown that adenovectors with modified capsid proteins, such as additions of polylysine or RGD sequence to fibers, can effectively infect cells with defective CAR expression [9][10][11]. Thus, modified adenovectors with increased transduction ability or targeting capacity may be useful for various gene transfer or gene therapy studies or clinical applications.…”

mentioning

confidence: 99%

A rapid and efficient method for purification of recombinant adenovirus with arginine–glycine–aspartic acid-modified fibers

Peng¹,

Wu²,

Davis³

et al. 2006

Analytical Biochemistry

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Recombinant adenoviral vectors (adenovectors) have been subject to various genetic modifications to improve their transduction efficiency and targeting capacity. Production and purification of adenovectors with modified capsid proteins can be problematic using conventional two-cycle CsCl gradient ultracentrifugation. We have developed a new method for purifying recombinant adenovectors in two steps: iodixanol discontinuous density gradient ultracentrifugation and sizeexclusion column chromatography. The purity and infectious activity of adenovectors isolated by either method were comparable. The new method yielded three to four times more adenovectors with RGD-modified fiber proteins than did the conventional CsCl method. For other fiber-modified and wild-type adenovectors, the yields of the two methods were comparable. Thus, the iodixanol-based method can be used not only to improve the production of RGD-modified adenovectors, but also to purify adenovectors with or without fiber modifications. Moreover, the whole procedure can be completed in 3 hours. Therefore, this method is rapid and efficient for production recombination adenovectors, especially those with RGD-modified fibers.Adenoviral vectors (adenovectors) have high in vivo transduction efficiencies and are easy to construct and produce. They are thus frequently used for in vitro and in vivo gene delivery and for gene therapy in clinical trials [1] . Substantial efforts have been made to characterize adenovirus-host cell interactions and to improve gene delivery by adenovectors, including minimizing vector gene expression, reducing vector related toxicity and immunogenicity, increasing their capacity to accommodate large foreign genes, and increasing their transduction efficiency.It is now known that adenovirus interacts with host cells by binding to the coxsackievirus and adenovirus receptor (CAR) [2], a cellular receptor for Ad5 and most other adenovirus serotypes. After attachment, the virion is then internalized by endocytosis through the interaction of its penton base with α v β 3 and α v β 5 integrins on the host cell surface [3]. However, there is growing evidence that the expression of CAR is frequently suppressed in various types of cells, including tumor cells and in primary tumors [4,5], resulting in resistance to adenovirus infection [6][7][8] Unfortunately, technical difficulties may be encountered in production of certain modified adenovectors. We have encountered difficulties when producing certain adenovectors by this method, especially those with RGD-modified fibers or "sticky" virus. For example, we have experienced difficulty in purifying various RGD-modified adenoviral vectors by conventional two-cycle (2x) of CsCl ultracentrifugation because the RGD-modified adenoviral vectors tend to aggregate, especially in the second round of ultracentrifugation, forming floccules without a sharp band. This results in a low yield or even failure of purification. Thus, an alternative method for efficient production of these modified adenov...

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“…Although ligands fused to the C terminus of the fiber molecule can physically interact with cognate cellular receptors, the C terminus of fiber can only accommodate ligands up to 30 aa in size, other than the non-structured biotin acceptor protein. 75,78,79 Alternatively, small ligands such as FLAG, RGD, polylysine and transferrin receptor binding peptides can be incorporated into the highly flexible HI loop of fiber. [31][32][33][34] Belousova and co-workers 37 have recently demonstrated the capacity of the HI loop to accommodate large (480 aa) targeting ligands by successfully generating infectious virions containing fragments of the penton base RGD loop inserted into the HI loop.…”

Section: Discussionmentioning

confidence: 99%

HER3 targeting of adenovirus by fiber modification increases infection of breast cancer cells in vitro, but not following intratumoral injection in mice

et al. 2012

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Despite the tremendous potential of adenovirus (Ad) as a delivery vector for cancer gene therapy, its use in clinical settings has been limited, mainly as a result of the limited infectivity in many tumors and the wide tissue tropism associated with Ad. To modify the tropism of the virus, we have inserted the epidermal growth factor-like domain of the human heregulin-a (HRG) into the HI loop of Ad5 fiber. This insertion had no adverse effect on fiber trimerization nor did it affect incorporation of the modified fiber into infectious viral particles. Virions bearing modified fiber displayed growth characteristics and viral yields indistinguishable from those of wild-type (wt) virus. Most importantly, HRG-tagged virions showed enhanced infection of cells expressing the cognate receptors HER3/ErbB3 and HER4/ErbB4. This was significantly reduced in the presence of soluble HRG. Furthermore, HER3-expressing Chinese hamster ovary (CHO) cells were transduced by the HRG-modified virus, but not by wt virus. In contrast, CHO cells expressing the coxsackie-Ad receptor were transduced with both viruses. However, infection of an in vivo breast cancer xenograft model after intratumoral injection was similar with both viruses, suggesting that the tumor microenvironment and/or the route of delivery have important roles in infection of target cells with fiber-modified Ads.Cancer Gene Therapy (2012) 2,3 In addition to being among the most extensively studied viral vectors, Ads can be easily and economically manipulated, maintained and propagated to produce high titer stocks. 4,5 Moreover, they have the capacity to carry relatively large fragments of exogenous DNA into a wide range of cell and tissue types. 4,5 Subgroup C Ads (exemplified by the most widely used Ad5 and Ad2) attach to and enter host cells in a two-step process. 6 The initial recognition/attachment step is mediated by interactions between the knob component of fiber protein on the Ad capsid and the extracellular domain of the coxsackieadenovirus receptor (CAR) on the host cell surface. 7-11 This attachment is followed by a second interaction between the RGD motif in the penton base of the virus capsid and a v b 3 or a v b 5 cell surface integrins (secondary Ad receptors), which allows viral entry via receptor-mediated endocytosis. [12][13][14][15] Both the primary and secondary Ad receptors are expressed on a wide range of tissues leading to the observed broad tissue tropism of Ads. 16,17 Unfortunately, a large number of tumor cells appear relatively refractory to Ad infection because of limited surface expression of the primary Ad receptor. 1,[18][19][20][21][22][23] This observation has been a driving force behind recent intensive efforts to generate Ad viral vectors with altered tropism.

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Adenovirus targeted to heparan-containing receptors increases its gene delivery efficiency to multiple cell types

Cited by 301 publications

References 18 publications

Tumor-specific gene transfer via an adenoviral vector targeted to the pan-carcinoma antigen EpCAM

Tumor-specific gene transfer via an adenoviral vector targeted to the pan-carcinoma antigen EpCAM

A rapid and efficient method for purification of recombinant adenovirus with arginine–glycine–aspartic acid-modified fibers

HER3 targeting of adenovirus by fiber modification increases infection of breast cancer cells in vitro, but not following intratumoral injection in mice

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