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Targeting canine bladder transitional cell carcinoma with a human bladder cancer-specific ligand.

Lin TY, Zhang H, Wang S, Xie L, Li B, Rodriguez CO, de Vere White R, Pan CX - Mol. Cancer (2011)

Bottom Line: In vivo tumor-specific homing/targeting property and biodistribution of PLZ4 was performed in a mouse xenograft model via tail vein injection and was confirmed with ex vivo imaging.No significant changes in cell viability or proliferation were observed upon incubation with PLZ4.The in vivo and ex vivo optical imaging study showed that, when linked with the near-infrared fluorescent dye Cy5.5, PLZ4 substantially accumulated at the canine bladder cancer foci in the mouse xenograft model as compared to the control.

View Article: PubMed Central - HTML - PubMed

Affiliation: Division of Hematology and oncology, Department of Internal Medicine, University of California-Davis Cancer Center, Sacramento, CA 95817, USA.

ABSTRACT

Objective: To determine if a human bladder cancer-specific peptide named PLZ4 can target canine bladder cancer cells.

Experimental design: The binding of PLZ4 to five established canine invasive transitional cell carcinoma (TCC) cell lines and to normal canine bladder urothelial cells was determined using the whole cell binding assay and an affinitofluorescence assay. The WST-8 assay was performed to determine whether PLZ4 affected cell viability. In vivo tumor-specific homing/targeting property and biodistribution of PLZ4 was performed in a mouse xenograft model via tail vein injection and was confirmed with ex vivo imaging.

Results: PLZ4 exhibited high affinity and specific dose-dependent binding to canine bladder TCC cell lines, but not to normal canine urothelial cells. No significant changes in cell viability or proliferation were observed upon incubation with PLZ4. The in vivo and ex vivo optical imaging study showed that, when linked with the near-infrared fluorescent dye Cy5.5, PLZ4 substantially accumulated at the canine bladder cancer foci in the mouse xenograft model as compared to the control.

Conclusions and clinical relevance: PLZ4 can specifically bind to canine bladder cancer cells. This suggests that the preclinical studies of PLZ4 as a potential diagnostic and therapeutic agent can be performed in dogs with naturally occurring bladder cancer, and that PLZ4 can possibly be developed in the management of canine bladder cancer.

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Homing of PLZ4 to mouse xenograft of canine bladder cancers. Nude mice with the xenografts from TCC-PU-In at 0.5-0.8 cm in diameter were randomly selected to be injected with 100 μl (6 nmol) of pre-incubated PLZ4-biotin-SA-Cy5.5 complex or SA-Cy5.5 dye as the control. Total body imaging was performed at 0, 1, 3, 6, and 12 hours after injection. All experiments were conducted in compliance with institutional guidelines and according to the protocol (No. 12988) approved by the Institutional Animal Care and Use Committee of the University of California at Davis. A. In vivo imaging of canine K9TCC-PU-In xenografts with PLZ4. In vivo near-infrared fluorescence images were taken at different time points after injection. C: the control mouse that received SA-Cy5.5. PLZ4: the mouse that received PLZ4-Cy5.5. Red arrows point to tumor xenografts. B. Ex vivo imaging of organs and tumor xenografts for fluorescence intensity. A color bar with the fluorescence intensity in arbitrary units is shown at the bottom. C. Ex vivo quantitative analysis of fluorescence uptake in tumor xenografts. The fluorescence intensity of tumor xenografts was normalized to that of the liver and kidney of the same mouse (the normalized value of liver and kidney is defined as 1.0).
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Figure 3: Homing of PLZ4 to mouse xenograft of canine bladder cancers. Nude mice with the xenografts from TCC-PU-In at 0.5-0.8 cm in diameter were randomly selected to be injected with 100 μl (6 nmol) of pre-incubated PLZ4-biotin-SA-Cy5.5 complex or SA-Cy5.5 dye as the control. Total body imaging was performed at 0, 1, 3, 6, and 12 hours after injection. All experiments were conducted in compliance with institutional guidelines and according to the protocol (No. 12988) approved by the Institutional Animal Care and Use Committee of the University of California at Davis. A. In vivo imaging of canine K9TCC-PU-In xenografts with PLZ4. In vivo near-infrared fluorescence images were taken at different time points after injection. C: the control mouse that received SA-Cy5.5. PLZ4: the mouse that received PLZ4-Cy5.5. Red arrows point to tumor xenografts. B. Ex vivo imaging of organs and tumor xenografts for fluorescence intensity. A color bar with the fluorescence intensity in arbitrary units is shown at the bottom. C. Ex vivo quantitative analysis of fluorescence uptake in tumor xenografts. The fluorescence intensity of tumor xenografts was normalized to that of the liver and kidney of the same mouse (the normalized value of liver and kidney is defined as 1.0).

Mentions: Next, we determined the tumor-specific homing/targeting property and in vivo biodistribution/binding specificity of PLZ4 in a mouse model with canine TCC xenografts. The near-infrared fluorescent dye Cy5.5 was used for in vivo and ex vivo imaging. PLZ4-biotin-SA-Cy5.5 (PLZ4-Cy5.5) complex or SA-Cy5.5 dye (control), 100 μl (6 nmol) for each mouse, was injected via tail vein into mice carrying TCC-PU-In xenografts at 0.5-0.8 cm in diameter. Substantial accumulation of signals was observed at the tumor site in the mice injected with PLZ4-Cy5.5 complex in a time-dependent manner with a maximum signal observed at 12 hours (Figure 3A). In contrast, negligible fluorescence uptake of Cy5.5 dye by tumors was detected in the control mice receiving SA-Cy5.5. To determine if there was any non-specific uptake of the dye by other vital organs, mice were euthanized at 12 hours after injection, vital organs and cancer xenografts were removed for ex vivo imaging (Figure 3B). Both liver and kidney demonstrated considerable signals even in the control mice that received SA-Cy5.5, suggesting the non-specific uptake. Compared with the tumor xenografts from the control mice treated with SA-Cy5.5, xenografts from the mice that received PLZ4-Cy5.5 accumulated significantly higher fluorescence signals after normalization with the fluorescence signal in liver (3.2 times, p = 0.003) and kidney (3.8 times, p < 0.001) (Figure 3C). No significant fluorescence uptake was observed in other organs including bladder. Collectively, these data demonstrated that PLZ4 exhibited excellent homing property toward TCC xenografts in vivo.


Targeting canine bladder transitional cell carcinoma with a human bladder cancer-specific ligand.

Lin TY, Zhang H, Wang S, Xie L, Li B, Rodriguez CO, de Vere White R, Pan CX - Mol. Cancer (2011)

Homing of PLZ4 to mouse xenograft of canine bladder cancers. Nude mice with the xenografts from TCC-PU-In at 0.5-0.8 cm in diameter were randomly selected to be injected with 100 μl (6 nmol) of pre-incubated PLZ4-biotin-SA-Cy5.5 complex or SA-Cy5.5 dye as the control. Total body imaging was performed at 0, 1, 3, 6, and 12 hours after injection. All experiments were conducted in compliance with institutional guidelines and according to the protocol (No. 12988) approved by the Institutional Animal Care and Use Committee of the University of California at Davis. A. In vivo imaging of canine K9TCC-PU-In xenografts with PLZ4. In vivo near-infrared fluorescence images were taken at different time points after injection. C: the control mouse that received SA-Cy5.5. PLZ4: the mouse that received PLZ4-Cy5.5. Red arrows point to tumor xenografts. B. Ex vivo imaging of organs and tumor xenografts for fluorescence intensity. A color bar with the fluorescence intensity in arbitrary units is shown at the bottom. C. Ex vivo quantitative analysis of fluorescence uptake in tumor xenografts. The fluorescence intensity of tumor xenografts was normalized to that of the liver and kidney of the same mouse (the normalized value of liver and kidney is defined as 1.0).
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Figure 3: Homing of PLZ4 to mouse xenograft of canine bladder cancers. Nude mice with the xenografts from TCC-PU-In at 0.5-0.8 cm in diameter were randomly selected to be injected with 100 μl (6 nmol) of pre-incubated PLZ4-biotin-SA-Cy5.5 complex or SA-Cy5.5 dye as the control. Total body imaging was performed at 0, 1, 3, 6, and 12 hours after injection. All experiments were conducted in compliance with institutional guidelines and according to the protocol (No. 12988) approved by the Institutional Animal Care and Use Committee of the University of California at Davis. A. In vivo imaging of canine K9TCC-PU-In xenografts with PLZ4. In vivo near-infrared fluorescence images were taken at different time points after injection. C: the control mouse that received SA-Cy5.5. PLZ4: the mouse that received PLZ4-Cy5.5. Red arrows point to tumor xenografts. B. Ex vivo imaging of organs and tumor xenografts for fluorescence intensity. A color bar with the fluorescence intensity in arbitrary units is shown at the bottom. C. Ex vivo quantitative analysis of fluorescence uptake in tumor xenografts. The fluorescence intensity of tumor xenografts was normalized to that of the liver and kidney of the same mouse (the normalized value of liver and kidney is defined as 1.0).
Mentions: Next, we determined the tumor-specific homing/targeting property and in vivo biodistribution/binding specificity of PLZ4 in a mouse model with canine TCC xenografts. The near-infrared fluorescent dye Cy5.5 was used for in vivo and ex vivo imaging. PLZ4-biotin-SA-Cy5.5 (PLZ4-Cy5.5) complex or SA-Cy5.5 dye (control), 100 μl (6 nmol) for each mouse, was injected via tail vein into mice carrying TCC-PU-In xenografts at 0.5-0.8 cm in diameter. Substantial accumulation of signals was observed at the tumor site in the mice injected with PLZ4-Cy5.5 complex in a time-dependent manner with a maximum signal observed at 12 hours (Figure 3A). In contrast, negligible fluorescence uptake of Cy5.5 dye by tumors was detected in the control mice receiving SA-Cy5.5. To determine if there was any non-specific uptake of the dye by other vital organs, mice were euthanized at 12 hours after injection, vital organs and cancer xenografts were removed for ex vivo imaging (Figure 3B). Both liver and kidney demonstrated considerable signals even in the control mice that received SA-Cy5.5, suggesting the non-specific uptake. Compared with the tumor xenografts from the control mice treated with SA-Cy5.5, xenografts from the mice that received PLZ4-Cy5.5 accumulated significantly higher fluorescence signals after normalization with the fluorescence signal in liver (3.2 times, p = 0.003) and kidney (3.8 times, p < 0.001) (Figure 3C). No significant fluorescence uptake was observed in other organs including bladder. Collectively, these data demonstrated that PLZ4 exhibited excellent homing property toward TCC xenografts in vivo.

Bottom Line: In vivo tumor-specific homing/targeting property and biodistribution of PLZ4 was performed in a mouse xenograft model via tail vein injection and was confirmed with ex vivo imaging.No significant changes in cell viability or proliferation were observed upon incubation with PLZ4.The in vivo and ex vivo optical imaging study showed that, when linked with the near-infrared fluorescent dye Cy5.5, PLZ4 substantially accumulated at the canine bladder cancer foci in the mouse xenograft model as compared to the control.

View Article: PubMed Central - HTML - PubMed

Affiliation: Division of Hematology and oncology, Department of Internal Medicine, University of California-Davis Cancer Center, Sacramento, CA 95817, USA.

ABSTRACT

Objective: To determine if a human bladder cancer-specific peptide named PLZ4 can target canine bladder cancer cells.

Experimental design: The binding of PLZ4 to five established canine invasive transitional cell carcinoma (TCC) cell lines and to normal canine bladder urothelial cells was determined using the whole cell binding assay and an affinitofluorescence assay. The WST-8 assay was performed to determine whether PLZ4 affected cell viability. In vivo tumor-specific homing/targeting property and biodistribution of PLZ4 was performed in a mouse xenograft model via tail vein injection and was confirmed with ex vivo imaging.

Results: PLZ4 exhibited high affinity and specific dose-dependent binding to canine bladder TCC cell lines, but not to normal canine urothelial cells. No significant changes in cell viability or proliferation were observed upon incubation with PLZ4. The in vivo and ex vivo optical imaging study showed that, when linked with the near-infrared fluorescent dye Cy5.5, PLZ4 substantially accumulated at the canine bladder cancer foci in the mouse xenograft model as compared to the control.

Conclusions and clinical relevance: PLZ4 can specifically bind to canine bladder cancer cells. This suggests that the preclinical studies of PLZ4 as a potential diagnostic and therapeutic agent can be performed in dogs with naturally occurring bladder cancer, and that PLZ4 can possibly be developed in the management of canine bladder cancer.

Show MeSH
Related in: MedlinePlus