• Molecules and Cells
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• 제25권4호
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• pp.462-466
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• 2008

Adenoviruses are the most commonly used gene-delivery vectors due to the efficiency of their in vivo gene transfer. Since 1993, about 300 protocols using an adenoviral vector have been performed, although they have yet to be proven effective in clinical trials. The adenovirus-based vector has been continuously improved by modification of the adenoviral genome and capsid, and novel adenovirus-delivery systems, such as the carrier-cell delivery system, have been recently proposed. Adenovirus-based cancer gene therapy is fast becoming one component of a multi-modality treatment approach to advanced cancer, along with surgery, radiotherapy, and chemotherapy.

• Toxicological Research
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• 제20권2호
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• pp.109-116
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• 2004

For the safety evaluation of adenovirus-mediated gene transfer, we investigated differential gene expressions after transfecting adenoviral vector containing p16 tumor suppressor gene (Ad5CMV-p16) into human non-small cell lung cancer cells. In the previous study, we showed adenovirus-mediated $p16^{INK4a}$ gene transfer resulted in significant inhibition of cancer cell growth. We investigated gene expression changes after transfecting Ad5CMV-p16, Ad5CMV (null type, a mock vector) into A549 cells by using cDNA chip and oligonucleotide microarray chip (1200 genes) which carries genes related with signal transduction pathways, cell cycle regulations, oncogenes and tumor suppressor genes. We found that $p16^{INK4a}$ gene transfer down regulated 5 genes (cdc2, cyclin D3, cyclin B, cyclin E, cdk2) among 26 genes involved in cell cycle regulations. Compared with serum-free medium treated cells, Ad5CMV-p16 changed 27 gene expressions, two fold or more on oligonucleotide chip. In addition, Ad5CMV-p16 did not seem to increase the tumorigenicity-related gene expression in A549 cells. Further studies will be needed to investigate the effect of Ad5CMV-p16 on normal human cells and tissues for safety evaluation.

• Environmental Analysis Health and Toxicology
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• 제24권1호
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• pp.71-77
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• 2009

Adenoviral vector(AdV) has been the most widely used viral vector for delivering an exogenous therapeutic gene to human. As of this date, more clinical trials utilize recombinant AdV to treat cancer and monogenic inherited disease as well as vaccine applications. However, the number of clinical trials had dropped markedly following the tragic death of a patient ongoing an AdV therapy for the treatment of an ornithine transcarbamylase deficiency(OTCD). This review is an attempt to provide the information on toxicity generated by AdVmediated gene transfer. It would serve as a sobering reality to researchers and clinicians exploring the use of AdV, as to the complications involved in human application.

• 한국독성학회:학술대회논문집
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• 한국독성학회 2002년도 Molecular and Cellular Response to Toxic Substances
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• pp.196-196
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• 2002

p16INK4a tumor suppressor gene transfer in the non-small cell lung cancer cells by transduction of recombinant adenovirus (Ad5CMV-p16) resulted in significant inhibition of cancer cell growth (Anticancer Res., 1998, 18:3257-3261). As a safety concern, we have investigated gene and protein expression after transduction of adenoviral vector (Ad5CMV-p16) in human non-small cell lung cancer (A549) cells by using microarray and 2D gel electrophoresis/ MALDI-TOF.(omitted)

• 미생물학회지
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• 제42권1호
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• pp.73-76
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• 2006

신정전구세포를 이용한 퇴행성 뇌질환의 세포치료나 유전자치료에서 효율적인 유전자 전달을 목적으로 개량된 아데노바이러스 벡터의 실용 가능성을 쥐의 해마에서 유래된 신정전구세포를 이용하여 조사하였다. 외피단백질을 조작한 개략 아데노바이러스벡터는 분화전과 후의 신정전구세포로 1세대 아데노바이러스 벡터에 비해 6배 정도 유전자를 효율적으로 전달하였다. 또한 바이러스의 감염은 신정전구세포가 신경세포나 신정 아교세포로 분화하는데 영향을 미치는 않았다. 따라서 신정전구세포를 이용한 신정질환의 세포치료나 유전자 치료에서 개량된 아데노바이러스로 유전자를 전달하면 치료의 효율성을 향상시킬 수 있을 것이다.

• IMMUNE NETWORK
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• 제6권4호
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• pp.192-198
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• 2006

Background: Investigating strategy to enhance efficiency of gene transfer via adenovirus is critical to sustain gene expression in targeted cells or tissues to regulate immune responses. However, the use of adenovirus as a gene delivery method has been limited by the native tropism of the virus. In this study, the critical parameter is to improve the efficient binding of viral particles to the plasma membrane prior to cellular uptake. Methods: Human immunodeficiency virus (HIV-1) trans-acting activator of transcription (TAT), a protein transduction domain, was fused to the ectodomain of the coxsackie-adenovirus receptor (CAR). The CAR-TAT protein was produced from a Drosophila Schneider 2 cells (S2) transfected with CAR-TAT genes. The function of CARTAT was analyzed the efficiency of adenoviral gene transfer by flow cytometry, and then immunizing AdVGFP with CAR-TAT was transduced on dendritic cells (DCs). Results: S2 transfectants secreting CAR-TAT fusion protein has been stable over a period of 6 months and its expression was verified by western blot. Addition of CAR-TAT induced higher transduction efficiency for AdVGFP at every MOI tested. When mice were vaccinated with DC of which adenoviral transduction was mediated by CAR-TAT, the number of IFN-${\gamma}$ secreting T-cells was increased as compared with those DCs transduced without CAR-TAT. Conclusion: Our data provide evidence that CAR-TAT fusion protein enhances adenoviral transduction and immunogenecity of transgenes on DCs and may influence on the development of adenoviral-mediated anti-tumor immunotherapy.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• 제32권4호
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• pp.308-316
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• 2006

Objectives Despite considerable advances in technique, experience and skill, the precise place of surgery in the treatment of facial nerve injury remains uncertain. We designed a facial nerve crush injury model in rats and evaluated the recovery of crushed nerve which is the most common injury type of facial nerve using adenovirus vector mediated in vivo gene transfer of Brain derived neurotrophic factor(BDNF). Materials and methods In 48 Sprague Dawley rats, we made a facial nerve crush injury model to main trunk before the furcation, and injected a $10^{11}$pfu adenoviral BDNF in experimental group(BDNF adenoviral injection group; ad-BDNF) and $3{\mu}l$ saline in control group(Saline injection group; saline). After a period of regeneration from 10 to 40 days, nerve regeneration was evaluated with functioinal test (vibrissae and ocular movement), electrophysiologic study(threshold, peak voltage, conduction velocity) and histomorphometric study of axon density. Results Vibrissae and ocular movement, threshold and conduction velocity improved as time elapse in both group, however axon density was increased significantly only in experimental group. Functional test in 10 days and 20 days showed no difference between experimental group and control group. Vibrissae movement, threshold, conduction velocity and axon density in 30 days revealed that the regeneration in quality of experimental group was significantly superior to that of control group. Conclusion In general, there is tendency for nerve regeneration in experimental group (BDNF-adenovirus injection group) during 40 days, functional recovery was detected successfully after facial nerve crush in 30 days postoperatively.

• 약학회지
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• 제57권4호
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• pp.265-271
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• 2013

Treatments of vascular disease via modulating the expression of specific proteins by gene transfer have been attempted in various studies over the past few years. Among several methods to deliver genes, adenovirus currently has been used because of a number of positive aspects. In this study, we test adenoviral vector as a potential mediator in the treatment of vascular disease by using freshly isolated vascular tissues not cultured vascular cells. Freshly isolated vascular tissues were directly exposed to adenoviral vector pAd5CMVmcsIRESeGFPpA to check the possibility of GFP expression in different layer of vascular tissues. We found that the GFP expression by using adenoviral vector experiments is mainly focused on the adventitia and failed to detect GFP expression at endothelial layer or vascular smooth muscle layer in vascular tissues. However, we also found that several integrin receptors are robustly expressed in vascular smooth muscle, thus the limited expression of protein in vascular smooth muscle are not likely the lack of integrin receptors. In conclusion, adenovirus could not be a good tool for a specific protein expression in vascular smooth muscle cell. Thus, the application of adenovirus as a tool for gene therapy of vascular smooth muscle cells in clinical therapeutic trial need to be optimized further.

• Journal of Korean Neurosurgical Society
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• 제30권sup1호
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• pp.13-19
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• 2001

Objective : We investigated the feasibility of a double suicide gene/prodrug therapy, involving direct introduction of the herpes simplex virus Type 1 thymidine kinase(TK) gene and the Escherichia coli cytosine deaminase(CD) gene, via a recombinant adenoviral vector and ganciclovir(GCV) and/or 5-fluorocytosine(5-FC) treatment, in C6 glioma cells. Methods : Efficient gene transfer and transduction of C6 glioma cells via a recombinant adenovirus were evaluated by infecting cells with adenovirus bearing the ${\beta}$-galactosidase gene and then staining cells with 5-bromo-4-chloro-3-indolyl-13-D-galactoside. CD/TK expression in cells infected with adenovirus bearing the CD/TK gene(ad-CD/TK) was examined by immunoblotting analysis. For in vitro cytotoxicity experiments, the cells were infected with ad-CD/TK or ad-${\Delta}E1$(as a control). After addition of a variety of concentrations of GCV and 5-FU, either separately or in combination, cell viability was determined by staining the cells with crystal violet solution 6 days after infection. Result : C6 glioma cells were efficiently transduced with recombinant adenoviral vector at multiplicities of infection of 200 or more. In vitro cytotoxicity of GCV and/or 5-FC, either alone or in combination, was exclusively observed in the cells transduced with ad-CD/TK. Obvious cytotoxicity(>50% inhibition) was observed in the presence of 5-FC at concentrations greater than 30ug/ml or GCV at concentrations greater than 0.3ug/ml at a multiplicity of infection of 100. Additionally, cytotoxicity in the presence of both GCV and 5-FC was greater than that after sinlge-prodrug treatments, indicating additive effects of the prodrug treatments. Conclusion : The administration of a double-suicide gene/prodrug therapy might have great potential in the treatment of brain tumors.

• 대한약학회:학술대회논문집
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• 대한약학회 2003년도 Proceedings of the Convention of the Pharmaceutical Society of Korea Vol.2-2
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• pp.99.2-99.2
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• 2003

One of the major limitations in using adenoviral vector for gene therapy is inefficient infection of host cells. The presence of coxsackievirus and adenovirus receptor (CAR) and ${\alpha}$-integrin on cell surfaces is required for efficient adenovirus infection. In this study, we investigated the effect of trichostatin A, a histone deacetylase inhibitor, on transfection efficiency after transduction of adenovirus mediated p16$\^$INK4a/ gene transfer. In our previous study, p16$\^$INK4a/ tumor suppressor gene transfer in the non-small cell lung cancer cells (A549 cells) by transduction of recombinant adenovirus (Ad5CMV-p16) resulted in significant inhibition of cancer cell proliferation. (omitted)