Adenovirus type 5 uptake by lung adenocarcinoma cells in culture correlates with Ad5 fibre binding is mediated by ?v?1 integrin and can be modulated by changes in ?1 integrin function

Davison, Elizabeth; Kirby, Ian; Whitehouse, J.L.; Hart, Ian R.; Marshall, John F.; Santis, George

doi:10.1002/jgm.223

Cited by 48 publications

(33 citation statements)

References 34 publications

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“…This process requires the presence of both primary and secondary receptors for the Ad5 virion (i.e., CAR and a 3 b V or a 5 b V integrins) on the cell surface at levels sufficient to allow binding by the vector. 46,47 As most of the clinical tumor cell isolates are heterogeneous, we have tested all our cell lines for susceptibility to Ad5 vector infection by titrating the replication competent rcAd/ CMV-GFP vector on each of the cell lines (Fig 4). The ability of rcAd/CMV-GFP to replicate in all human cell lines allows for an easy detection of transduced cells, since vector genome amplification boosts GFP expression levels many fold.…”

Section: Bt Cell Lines Are Efficiently Transduced By Ad5-based Vectorsmentioning

confidence: 99%

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Enhanced apoptosis of glioma cell lines is achieved by co-delivering FasL-GFP and TRAIL with a complex Ad5 vector

Rubinchik

Woraratanadharm

et al. 2003

Cancer Gene Ther

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Brain tumors (BTs) are among the most malignant forms of human cancer. Unfortunately, current treatments are often ineffective and produce severe side effects. Cytotoxic gene therapy is an alternative treatment strategy, with the potential advantages of reduced toxicity to normal brain tissue. Apoptosis-inducing ''death ligands'' Fas ligand and TNF-related apoptosis-inducing ligand (TRAIL) are genes with substantial cytotoxic activity in susceptible tumor cells. Here, we compared the effectiveness of Ad vectormediated delivery of Fas ligand-green fluorescent protein (FasL-GFP) fusion protein, human TRAIL, and both genes simultaneously. We examined a panel of 13 cell lines (eight derived from primary isolates) for susceptibility to Ad5-based vector infection and for sensitivity to FasL-and TRAIL-mediated apoptosis. All cell lines were efficiently transduced, but, as expected, varied in their sensitivity to ligand-induced apoptosis. Generally, sensitivity to FasL-GFP correlated with cell surface FasR levels, but no such correlation was seen for TRAIL and its functional receptors, DR4 and DR5. The vector expressing both FasL-GFP and TRAIL was more effective than either of the single-gene vectors at comparable transduction levels, and it was effective against a broader range of cell lines. In five cell lines, coexpression resulted in apoptosis levels greater than those predicted for strictly additive activity of the two death ligands. We believe that Ad vector-mediated delivery of multiple death ligands may be developed as a potential BT therapy, either alone or in conjunction with surgical resection of the primary tumor.

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Section: Bt Cell Lines Are Efficiently Transduced By Ad5-based Vectorsmentioning

confidence: 99%

“…46,47 We determined cell surface levels of FasR and DR4 and DR5 (functional TRAIL receptors) for each cell line using flow cytometry (Fig 5). The majority of cell lines had levels of FasR comparable to or greater than those of HeLa.…”

Section: Death Ligand Gene Therapy Of Brain Tumor Cells S Rubinchik Ementioning

confidence: 99%

Enhanced apoptosis of glioma cell lines is achieved by co-delivering FasL-GFP and TRAIL with a complex Ad5 vector

Rubinchik

Woraratanadharm

et al. 2003

Cancer Gene Ther

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“…A strong correlation between the expression of CAR on the cell surface and the level of transgene expression in photochemically treated cells was observed, suggesting that the absolute level of virus binding to the cell surface determines the level of transduction in photochemically treated cells, as reported by other groups for conventional adenoviral infection. [18][19][20] Blocking CAR with recombinant Ad5 fiber-knob resulted in a complete inhibition of gene expression, both in conventionally infected and in photochemically treated cells. These results strengthen the assumption that attachment to the cell surface is a key event for PCI-mediated transduction and suggest further that nonreceptor-mediated uptake pathways are not involved in the photochemical transduction pathway.…”

Section: Discussionmentioning

confidence: 99%

“…Using the RmcB anti-CAR monoclonal antibody, 13 we found that the cell lines used in this study expressed different levels of CAR ( Table 1) Fig 1b), as previously reported by other groups. [18][19][20] The relationship between the expression of CAR and the percentage of b-galactosidasepositive cells in photochemically treated cells showed an even better linear correlation (P ¼ .002 , Fig 1b), suggesting that virus binding to the cell surface is of fundamental importance also for photochemical transduction.…”

Section: Influence Of Car On Photochemical Transductionmentioning

confidence: 99%

PCI-enhanced adenoviral transduction employs the known uptake mechanism of adenoviral particles

et al. 2005

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The development of methods for efficient and specific delivery of therapeutic genes into target tissues is an important issue for further development of in vivo gene therapy. In the present study, the physical targeting technique, photochemical internalization (PCI), has been used together with adenovirus. The combination of PCI and adenoviral transduction has previously been shown to be favorable compared to adenovirus used alone, and the aim of this study was to verify the role of the adenoviral receptors and identify the uptake pathway used by adenoviral particles in photochemically treated cells. All examined cell lines showed augmented transduction efficiency after PCI-treatment, with a maximum of 13-fold increase in transgene expression compared to conventionally infected cells. Blocking of CAR induced a complete inhibition of PCI-enhanced transgene expression. However, photochemical treatment managed to enhance the transduction efficiency of the retargeted virus AdRGD-GFP showing also that the virus-CAR interaction is not vital for obtaining a photochemical effect on adenoviral transduction. Blocking the a V -integrins reduced the gene expression significantly in photochemically treated cells. Subjecting HeLa cells expressing negative mutant-dynamin to light treatment after infection gave no significant increase in gene transfer, while the gene transfer were enhanced seven-fold in cells with wild-type dynamin. Furthermore, chlorpromazine inhibited photochemical transduction in a dose-dependent manner, whereas Filipin III had no effect on the gene transfer. In summary, the data presented imply that adenoviral receptor binding is important and clathrin-mediated endocytosis is the predominant uptake mechanism for adenoviral particles in photochemically treated cells.

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“…The human non-small cell carcinoma cell line, A549, is widely used to study the role of integrins in normal and pathophysiological processes (36)(37)(38)(39)(40)(41)(42)(43)(44). We validated the presence of ABI in this tumor model by Western blot and immunohistochemistry (IHC).…”

Section: A549 Tumor Cells Express Abimentioning

confidence: 99%

Synergistic effects of light-emitting probes and peptides for targeting and monitoring integrin expression

Achilefu

Bloch²,

Markiewicz³

et al. 2005

Proc. Natl. Acad. Sci. U.S.A.

119

113

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mouse ͉ optical imaging ͉ RGD peptides ͉ tumor ͉ near-infrared A ngiogenesis, the formation of new blood vessels, is the cardinal feature of virtually all malignant tumors (1). Because of this commonality, probing tumor-induced angiogenesis and associated proteins is a viable approach to detect and treat a wide range of cancers. Angiogenesis is stimulated by integrins, a large family of transmembrane proteins that mediate dynamic linkages between extracellular adhesion molecules and the intracellular actin skeleton. Integrins are composed of two different subunits, ␣ and ␤, which are noncovalently bound into ␣␤ complexes (2-4). Particularly, the expression of ␣ v ␤ 3 integrin (ABI) in tumor cells undergoing angiogenesis and on the epithelium of tumor-induced neovasculature alters the interaction of cells with the extracellular matrix, thereby increasing tumorigenicity and invasiveness of cancers (5-9).Numerous studies have shown that ABI and more than seven other heterodimeric integrins recognize proteins and low molecular weight ligands containing RGD (arginine-glycineaspartic acid) motifs in proteins and small peptides (10). Based on structural and bioactivity considerations, cyclic RGD peptide ligands are preferentially used as delivery vehicles for molecular probes for imaging (8,(11)(12)(13) and treating (14-17) ABI-positive tumors and proliferating blood vessels. Until recently, most of the in vivo imaging studies were performed with radiopharmaceuticals because of the high sensitivity and clinical utility of nuclear imaging methods. Particularly, the use of small monoatomic radioisotopes does not generally interfere with the biodistribution and bioactivity of ligands. Despite these advantages, nuclear imaging is currently only performed in specialized centers because of regulatory, production, and handling issues associated with radiopharmaceuticals. Optical imaging is an alternative but complementary method to interrogate molecular processes in vivo and in vitro.Optical imaging for biomedical applications typically relies on activating chromophore systems with low energy radiation between 400 -and 1,500-nm wavelengths and monitoring the propagation of light in deep tissues with a charge-coupled device camera or other point source detectors (18). Molecular optical imaging of diseases with molecular probes is attractive because of the flexibility of altering the detectable spectral properties of the probes, especially in the fluorescence detection mode. The probes can be designed to target cellular and molecular processes at functional physiological concentrations. For deep-tissue imaging, molecular probes that are photoactive in the near-infrared (NIR) instead of visible wavelengths are preferred to minimize background tissue autof luorescence and light attenuation caused by absorption by intrinsic chromophores (19). In contrast to radioisotopes, the NIR antennas are usually large heteroatomic molecules that could impact the biodistribution and activity of conjugated bioactive ligands. However, previous s...

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Adenovirus type 5 uptake by lung adenocarcinoma cells in culture correlates with Ad5 fibre binding is mediated by ?v?1 integrin and can be modulated by changes in ?1 integrin function

Cited by 48 publications

References 34 publications

Enhanced apoptosis of glioma cell lines is achieved by co-delivering FasL-GFP and TRAIL with a complex Ad5 vector

Enhanced apoptosis of glioma cell lines is achieved by co-delivering FasL-GFP and TRAIL with a complex Ad5 vector

PCI-enhanced adenoviral transduction employs the known uptake mechanism of adenoviral particles

Synergistic effects of light-emitting probes and peptides for targeting and monitoring integrin expression

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