An improved intravesical model using human bladder cancer cell lines to optimize gene and other therapies

Watanabe, Takafumi; Shinohara, Nobuo; Sazawa, Ataru; Harabayashi, Toru; Ogiso, Yoshifumi; Koyanagi, Tomohiko; Takiguchi, Mitsuyoshi; Hashimoto, Akira; Kuzumaki, Noboru; Yamashita, Motoyuki; Tanaka, Motoyoshi; Grossman, H. Barton; Benedict, William F.

doi:10.1038/sj.cgt.7700261

Cited by 62 publications

(52 citation statements)

References 10 publications

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“…7 Ad-IFNa2a1 12 and AdIFNa2b 13 are both E1-deleted adenoviral constructs with the E1 region replaced with a CMV promoter driving the IFN transgene including the secretory leader sequence. Ad-IFNa2a1 is identical to Ad-IFNa2b except that the Ad-IFNa2a1 transgene is comprised of the cDNA that encodes the N-terminus of the human IFNa2b fused to the C-terminus of the human IFNa1 through a BglII restriction site ((Hu-IFN-aA/D (BglII)), whereas the AdIFNa2b contains the transgene for the human IFNa2b gene.…”

Section: Cell Lines Vectors and Syn3mentioning

confidence: 99%

“…The methods for growing superficial KU7/GFP human bladder tumors in athymic mice and their imaging have previously been described. 7,8 At 2 weeks after the tumor cells were intravesically instilled, the bladders were imaged for the presence of GFP-containing tumors. Anterior, posterior, as well as left lateral and right lateral views were taken as previously described.…”

Section: Superficial Tumor Formation Treatment and Imagingmentioning

confidence: 99%

“…To facilitate our efficacy studies, a recently developed superficial bladder tumor model was utilized to monitor the progression of human bladder tumors growing in athymic mice by GFP fluoresence. 7,8 In our initial study, we found that two daily 1 h intravesical administrations of Ad-IFNa at a concentration of 1 Â 10 11 particles/ml (P/ml), together with Syn3, produced a marked tumor regression in superficial human bladder tumors. 9 All other treatments including Syn3 alone, Ad-bgal/Syn3, Ad-IFN without Syn3, or the IFN protein alone at high concentration showed no effect on tumor growth.…”

Section: Introductionmentioning

confidence: 99%

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Efficacy of a single intravesical treatment with Ad-IFN/Syn 3 is dependent on dose and urine IFN concentration obtained: implications for clinical investigation

Tao

Connor²,

Ashoori

et al. 2005

Cancer Gene Ther

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There is a need to improve the treatment of superficial bladder cancer. One area which holds promise is intravesical gene therapy. Recently, studies undertaken by us have shown that marked tumor regression of bladder cancers occurred after two daily intravesical administrations of an adenovirus encoding human interferon a (Ad-IFNa) using a mouse superficial bladder cancer model in which human bladder tumors are growing. A dose of 1 Â 10 11 particles/ml (P/ml) was used along with 1 mg/ml of Syn3, a gene transfer-enhancing agent. Since clinical studies are being planned using this approach, it became critical to determine if one exposure and lower particle number could be equally effective. We report that indeed a single dose of Ad-IFNa in Syn3 at doses of 1 Â 10 10 -1 Â 10 11 P/ml is highly effective in reducing the size of the tumors, whereas 1 Â 10 9 P/ml was not. Efficacy was also correlated with the level of IFN produced in the urine after treatment. Based on the results of the present studies, a Phase I trial is being planned for superficial bladder cancer, which will involve a single initial treatment with Ad-IFNa/Syn3 and measurement of IFN in the urine over time as an indicator of adequate gene transfer and expression.

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Section: Cell Lines Vectors and Syn3mentioning

confidence: 99%

Section: Superficial Tumor Formation Treatment and Imagingmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Efficacy of a single intravesical treatment with Ad-IFN/Syn 3 is dependent on dose and urine IFN concentration obtained: implications for clinical investigation

Tao

Connor²,

Ashoori

et al. 2005

Cancer Gene Ther

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“…2 The cells were maintained in Earl's MEM containing 10% fetal bovine serum and 50 U / mL of penicillin and streptomycin.…”

Section: Human Bladder Cancer Cell Linementioning

confidence: 99%

“…2 In addition, our laboratory in collaboration with others has shown that this model can be used to document efficient intravesical adenoviral -mediated gene transfer when it is combined with the polyamide compound, Syn 3. 3 Our next challenge was to use this tumor model in vivo to examine the efficacy of various treatments and schedules.…”

mentioning

confidence: 99%

Visualizing superficial human bladder cancer cell growth in vivo by green fluorescent protein expression

et al. 2002

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There has been no reliable orthotopic model available to visualize the growth of human superficial bladder cancer over time and to evaluate the efficacy of intravesical therapies. We have developed a novel approach to accomplish this task by generating human superficial bladder tumor cells to stably express high levels of green fluorescent protein ( GFP ) in vivo. Superficial bladder tumors were produced in athymic mice by intravesical instillation. In our initial studies tumors were quantitated by image analysis at a single time point, and the results compared to the estimation of the percentage of GFP cells present using flow cytometry after obtaining single cell suspensions of normal and tumor cells in the same bladder. A high correlation between the two methods was seen. Therefore, in subsequent studies, approximately 1 week after the intravesical instillation of the GFP expressing cancer cells a small incision was made to expose the bladder. The anterior, posterior, and lateral images of each bladder were captured to visualize GFPexpressing tumors. The ratio of green fluorescence pixel area, which represented the tumor burden, to the total area of the bladder was then calculated. A similar procedure was performed at 2, 3, and 4 weeks after instillation of the tumor cells. Using this procedure tumor progression over time could be measured in each mouse. By using this approach, it will now be possible to monitor the initial tumor sizes in the bladder of each mouse and then to evaluate the efficacy of various intravesical therapy protocols including intravesical gene therapy alone or in combination with other treatment modalities. Cancer Gene Therapy ( 2002 ) 9, 681 -686 doi:10.1038/sj.cgt.7700489Keywords: bladder cancer; green fluorescent protein; image analysis B ladder cancer is the fifth most frequent cancer in the United States. More than 50,000 individuals are diagnosed with bladder cancer each year. Between 70% and 80% of these cases involve superficial tumor. Such lesions are usually managed with transurethral resection often followed by intravesical immunotherapy with Bacillus Calmette -Guerin ( BCG ). Although BCG can prolong the length of progression -free survival after the tumor is resected, approximately 50% of superficial lesions will continue to recur and as many as 30% will progress to a higher grade and /or stage. 1 Therefore, new therapeutic approaches, including gene therapy alone or in combination with other modalities of therapy, need to be considered for the treatment of refractory superficial bladder disease.We recently have developed an intravesical model to produce superficially growing human bladder tumors in athymic mice with high frequency. 2 In addition, our laboratory in collaboration with others has shown that this model can be used to document efficient intravesical adenoviral -mediated gene transfer when it is combined with the polyamide compound, Syn 3. 3 Our next challenge was to use this tumor model in vivo to examine the efficacy of various treatments and schedules. Howeve...

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Mouse Models of Human Bladder Cancer as a Tool for Drug Discovery

et al. 2010

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Muscle-invasive bladder cancer is a deadly condition in dire need of effective new treatments. This unit contains a description of mouse models suitable for the evaluation of potential new therapies. Included is a genetically engineered mouse model of bladder cancer generated by the delivery of an adenovirus expressing Cre recombinase into the bladder lumen. Also described is an orthotopic mouse model created by the instillation of human bladder tumor cells into the bladder lumen of immune deficient mice. Protocols are also provided on the use of these models for the preclinical evaluation of new chemical entities, with mTOR inhibitors shown as an example.

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An improved intravesical model using human bladder cancer cell lines to optimize gene and other therapies

Cited by 62 publications

References 10 publications

Efficacy of a single intravesical treatment with Ad-IFN/Syn 3 is dependent on dose and urine IFN concentration obtained: implications for clinical investigation

Efficacy of a single intravesical treatment with Ad-IFN/Syn 3 is dependent on dose and urine IFN concentration obtained: implications for clinical investigation

Visualizing superficial human bladder cancer cell growth in vivo by green fluorescent protein expression

Mouse Models of Human Bladder Cancer as a Tool for Drug Discovery

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