The pressing challenge for contemporary gene therapy is to deliver enough therapeutic genes to enough cancer cells in vivo. With the aim of improving viral distribution and tumor penetration, we explored the use of decorin to enhance viral spreading and tumor tissue penetration. We generated decorin-expressing replication-incompetent (dl-LacZ-DCNG, dl-LacZ-DCNQ and dl-LacZ-DCNK) and replication-competent (Ad-DE1B-DCNG, Ad-DE1B-DCNQ and Ad-DE1B-DCNK) adenoviruses (Ads). Point mutants of decorin gene (DCNG), DCNK and DCNQ, have a negative and moderate binding affinity to type-I collagen fibril, respectively. In both tumor spheroids and established solid tumors in vivo, tissue penetration potency of dl-LacZ-DCNG was greatly enhanced than those of dl-LacZ, dl-LacZ-DCNQ and dl-LacZ-DCNK, and this enhanced tissue penetration effect derived from decorinexpressing Ad was dependent on the binding affinity of decorin to collagen fibril. Expression of DCNG enhanced viral spread of replicating Ad, leading to improved tumor reduction and survival benefit. Moreover, the tumoricidal effects of Ad-DE1B-DCNQ and Ad-DE1B-DCNK were lessened, as the binding affinity to collagen was decreased, showing that the increased cancer cell cytotoxicity was driven by the action of decorin on extracellular matrix (ECM). Furthermore, Ad-DE1B-DCNG substantially decreased ECM components within the tumor tissue. Finally, intratumoral injection of Ad-DE1B-DCNG in primary tumor site greatly reduced the formation of B16BL6 melanoma cell pulmonary metastases in mice. Taken together, these data show the utility of decorin as a dispersion agent and highlight its utility and potential in improving the efficacy of replicating Ad-mediated cancer gene therapy.

Oncolytic adenoviral vectors are currently being developed as biologic anticancer agents. Coupling the lytic function of an oncolytic adenovirus (Ad) with its ability as a transgene delivery system represents a powerful extension of this methodology. A clear advantage is the amplification of a therapeutic gene, as replicating vectors would be able to infect and deliver the gene of interest to neighboring cells. Granulocyte-macrophage colony-stimulating factor (GM-CSF) is one of the most potent stimulators of a specific and long-lasting antitumor immunity and its important role in the maturation of antigen-presenting cells to induce T-cell activation has been well documented. Similarly, the B7 family has also been shown to play an integral role in mediating an antitumor response. Most tumor cells, however, lack the expression of these costimulatory molecules on their surface, thus escaping immune system recognition. To increase the antitumor effect of an oncolytic Ad, we have generated an E1B 55 kDa-deleted oncolytic adenoviral vector, YKL-GB, that expresses both GM-CSF and B7-1. The therapeutic efficacy of YKL-GB Ad was evaluated in immunocompetent mice bearing murine melanoma B16-F10 tumors. Significant inhibition of tumor growth was seen in mice treated with YKL-GB compared to those treated with the analogous vector, YKL-1. Moreover, YKL-GB oncolytic Ad demonstrated enhanced antitumor activity and higher incidences of tumor regression compared to a replication-incompetent Ad, dl-GB, which coexpresses GM-CSF and B7-1. Localized GM-CSF and B7-1 gene transfer also conferred long-lasting immunity against a tumor re-challenge. To establish that the observed antitumor effect is associated with the generation of a tumorspecific immune response, we carried out interferon-g enzymelinked immune spot assay. We observed that YKL-GB induced significantly higher immune cell activation than YKL-1. Furthermore, immunohistochemical studies demonstrated robust dendritic cells and CD4 + /CD8 + T-cell infiltration in these mice compared to the YKL-1-treated groups. In agreement with these results, splenocytes from tumor-bearing mice treated with YKL-GB expressed high levels of the costimulatory and activation molecules. These findings demonstrate the effectiveness of enhancing the immune response against tumors with an oncolytic Ad expressing both GM-CSF and B7-1 and provide a potential therapeutic strategy for the management of neoplasia.

We have observed the unoccupied partial Pd 4d density of states in Cu Pd alloys by bremsstrahlung isochromat spectroscopy (BIS). It shows the filling of the Pd 4d band upon alloying, which is consistent with the Pd L7ii absorption spectroscopy measurements. We have also determined the total charge transfer between Pd and Cu atoms from the binding energy shifts in the care-level x-ray photoelectron spectroscopy and the intensity of unoccupied Pd d states in the BIS spectra. This is compared with the result from the Pd Li&& absorption spectroscopy.

Cu (400 Å)/polyimide was mixed with 80 keV Ar+ and N2+ from 1.0×1015 to 2.0×1016 ions/cm2. The same processes were repeated for the Cu (400 Å)/Al (50 Å)/polyimide system which has Al as a buffer layer. The quantitative adhesion strength was measured by a standard scratch test. X-ray photoelectron spectroscopy was employed to investigate the change in the chemical bonds of the ion beam mixed polyimide substrate and the intermediate effects for the adhesion enhancement in Cu/Al/polyimide. Two distinct tendencies are observed in the adhesion strength: Cu/Al/polyimide is more adhesive than Cu/polyimide after ion beam mixing, and N2+ ions are more effective in the adhesion enhancement than Ar+. The formation of an interlayer compound of CuAl2O4 accounts for the former, while the latter is understood by the fact that N2+ ions produce more pyridinelike moiety, amide group and tertiary amine moiety which are known as adhesion promoters.

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