Transduction of yeast cytosine deaminase mediated by HIV-1 Tat basic domain into tumor cells induces chemosensitivity to 5-fluorocytosine

Lee, Hakjoo; Ryu, Jiyoon; Kim, Kyung-Ae; Lee, Kil Soo; Lee, Jae Young; Park, Jae-Bong; Park, Jinseu; Choi, Soo Young

doi:10.1038/emm.2004.6

Cited by 8 publications

(6 citation statements)

References 22 publications

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“…In previous studies this has been addressed by generation of viral vectors encoding fusions of suicide genes with the HIV Tat gene [12], the HSV VP22 gene [13][14][15][16][17], or by direct employment either of a (i) malaria circumsporozoite (CS)-BCDase fusion protein [18] or a (ii) HIV-1 Tat basic domain-YCDase fusion protein [19].…”

Section: Introductionmentioning

confidence: 99%

Protein transduction with bacterial cytosine deaminase fused to the TLM intercellular transport motif induces profound chemosensitivity to 5-fluorocytosine in human hepatoma cells

Hillemann

Brandenburg

Schmidt

et al. 2005

Journal of Hepatology

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Section: Introductionmentioning

confidence: 99%

Protein transduction with bacterial cytosine deaminase fused to the TLM intercellular transport motif induces profound chemosensitivity to 5-fluorocytosine in human hepatoma cells

Hillemann

Brandenburg

Schmidt

et al. 2005

Journal of Hepatology

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“…Disadvantages include the lack of target cell specificity, potential of immunogenicity, and our incomplete knowledge of the molecular mechanisms of protein transduction. Reports of the application of PTD in potential therapeutic development are increasing in various fields and in a variety of cell systems and include inhibition of apoptosis by transduction of anti-apoptotic Bcl- XL protein in explant cultured human chondrocytes and in human islet cells [22,23], induction of chemosensitivity by transduction of cytosine deaminase in human tumor cells [24], and inhibition of proinflammatory signaling by transduction of superreppressor IkB in Jurkat T cells [21]. Protein transduction therapy is also showing promise in in vivo models, such as in the reduction of inflammation in carageenan-induced pleurisy of Wistar rat and the reduction of pancreatic islet cell toxicity in streptozotocin-induced diabetic mice [25,26].…”

Section: Discussionmentioning

confidence: 99%

Transduction of Cu, Zn-superoxide dismutase mediated by an HIV-1 Tat protein basic domain into human chondrocytes

Kim

Park

et al. 2006

Arthritis Res Ther

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This study was performed to investigate the transduction of a full-length superoxide dismutase (SOD) protein fused to transactivator of transcription (Tat) into human chondrocytes, and to determine the regulatory function of transduced Tat-SOD in the inflammatory cytokine induced catabolic pathway. The pTat-SOD expression vector was constructed to express the basic domain of HIV-1 Tat as a fusion protein with Cu, Zn-SOD. We also purified histidine-tagged SOD without an HIV-1 Tat and Tat-GFP as control proteins. Cartilage samples were obtained from patients with osteoarthritis (OA) and chondrocytes were cultured in both a monolayer and an explant. For the transduction of fusion proteins, cells/explants were treated with a variety of concentrations of fusion proteins. The transduced protein was detected by fluorescein labeling, western blotting and SOD activity assay. Effects of transduced Tat-SOD on the regulation of IL-1 induced nitric oxide (NO) production and inducible nitric oxide synthase (iNOS) mRNA expression was assessed by the Griess reaction and reverse transcriptase PCR, respectively. Tat-SOD was successfully delivered into both the monolayer and explant cultured chondrocytes, whereas the control SOD was not. The intracellular transduction of Tat-SOD into cultured chondrocytes was detected after 1 hours, and the amount of transduced protein did not change significantly after further incubation. SOD enzyme activity increased in a dosedependent manner. NO production and iNOS mRNA expression, in response to IL-1 stimulation, was significantly down-regulated by pretreatment with Tat-SOD fusion proteins. This study shows that protein delivery employing the Tat-protein transduction domain is feasible as a therapeutic modality to regulate catabolic processes in cartilage. Construction of additional Tat-fusion proteins that can regulate cartilage metabolism favorably and application of this technology in in vivo models of arthritis are the subjects of future studies.

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“…To overcome this problem, suicide enzymes can be fused to cell-penetrating peptides (CPPs), which are defined by their ability to reach the cytoplasmic and/or nuclear compartments in live cells after internalization. In previous studies, this was addressed by the generation of fusions of suicide genes with the HIV Tat gene [1], the HSV VP22 gene [2], and HBV PreS2 translocation motif (TLM) [3]. E. coli cytosine deaminase (bCD; EC 3.5.4.1) is responsible for the conversion of cytosine to uracil and ammonia, providing an important mechanism for pyrimidine salvage in microbes [4].…”

Section: Introductionmentioning

confidence: 99%

Inhibition of Tumor Growth by Polyarginine-Fused Mutant Cytosine Deaminase

Wang

Zhang

Zhao

et al. 2014

Appl Biochem Biotechnol

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Gene-directed enzyme-prodrug therapy is a method whereby cancerous tumors are selectively eradicated with minimal impact to healthy tissue. Due to its thermostability, E. coli cytosine deaminase (bCD) is one of the most widely used enzyme-prodrug combinations. However, wild-type bCD (wtbCD) displays a relatively poor turnover of 5-fluorocytosine (5-FC), and also has low permeability as a hexamer macromolecule (∼ 300 kDa), like many other therapeutic proteins. To improve these shortcomings, site-specific mutagenesis was performed by infusing the bCD with R9, a typical and highly effective cell-penetrating peptide. The results obtained by flow cytometry and confocal microscopy showed that the R9 efficiently delivered the enhanced green fluorescent proteins (EGFP) into the human liver hepatocellular carcinoma (HepG2) cells, and gathered at the nucleus, while EGFP alone did not have this ability. The penetrating efficiency of R9-EGPF was time and dose dependent. The results obtained by Western blot showed that R9-bCD, but not bCD proteins alone, could be uptaken into HepG2 cells. In vitro experiments showed that polyarginine enhanced the cytotoxicity of bCD, and R9-bCDmut had a stronger cytotoxicity than R9-bCD proteins. In vivo experiments also showed that R9-bCD and R9-bCDmut could prolong the survival time of tumor mice for 8-10 days. Future therapeutic applications of cell-permeable R9-bCDmut fusion proteins together with a systemic administration of 5-FC prodrug could result in profound anti-tumor activities.

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Transduction of yeast cytosine deaminase mediated by HIV-1 Tat basic domain into tumor cells induces chemosensitivity to 5-fluorocytosine

Cited by 8 publications

References 22 publications

Protein transduction with bacterial cytosine deaminase fused to the TLM intercellular transport motif induces profound chemosensitivity to 5-fluorocytosine in human hepatoma cells

Protein transduction with bacterial cytosine deaminase fused to the TLM intercellular transport motif induces profound chemosensitivity to 5-fluorocytosine in human hepatoma cells

Transduction of Cu, Zn-superoxide dismutase mediated by an HIV-1 Tat protein basic domain into human chondrocytes

Inhibition of Tumor Growth by Polyarginine-Fused Mutant Cytosine Deaminase

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