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Updated: Aug 14, 2026
An Orthotopic Bladder Cancer Model for Gene Delivery Studies
Published on: December 1, 2013
Development And In Vitro Characterization Of Bladder Tumor Cell Targeted Lipid-Coated Polyplex For Dual Delivery Of
Shayak Samaddar1, Joshua Mazur1, Devin Boehm1
1Department of Chemistry, Purdue University, Bindley Bioscience Center, West Lafayette, Indiana 47906, USA.
Background:
Bladder cancer is the fourth most common cancer in men and eleventh most common in women. Combination therapy using a gene and chemotherapeutic drug is a potentially useful strategy for treating bladder cancer in cases where a synergistic benefit can be achieved successfully. This approach relies on developing drug combinations using carrier systems that can load both hydrophilic genes and hydrophobic drugs. Ideally, the formulation for carrier system should be free of traditional high shear techniques such as sonication and extrusion to reduce shear-induced nucleic acid strand breakage. Moreover, the system should be able to protect the nucleic acid from enzymatic attack and deliver it specifically to the tumor site.
Materials And Methods:
A dual payload carrier system that was formulated using a simple flow mixing technique to complex anionic plasmid (EGFP-NLS) using a cationic polymer (CD-PEI2.5kD) followed by coating of the polyplex using lipid membranes. The resulting lipid-coated polyplex (LCP) formulations are targeted to bladder cancer cells by employing a bacterial adhesive peptide sequence, RWFV, that targets the LCP to the tumor stroma for efficiently delivering reporter plasmid, EGFP-NLS and a model small molecule drug, pyrene, to the cancer cells.
Results:
Encapsulation efficiency of the peptide targeted carrier for the plasmid was 50% ± 0.4% and for pyrene it was 16% ± 0.4%. The ability of the targeted LCP to transfect murine bladder cancer cells was 4-fold higher than LCP bearing a scrambled peptide sequence. Fluorescence of cells due to pyrene delivery was highest after 4 hrs using targeted LCP. Finally, we loaded the peptide targeted LCP with anti-cancer agent, curcumin. The targeted formulation of curcumin resulted in only 45% viable cancer cells at a concentration of 5 µg/mL, whereas the empty and non-targeted formulations did not result any significant cell death.
Conclusion:
These results demonstrate the specificity of the targeting peptide sequence in engaging tumor cells and the utility of the developed carrier platform to deliver a dual payload to bladder tumor cells.
Insights
A novel dual-payload carrier system effectively targets bladder cancer cells. This lipid-coated polyplex delivers genes and drugs, showing significant anti-cancer activity and specific tumor cell engagement.
Area of Science:
- Biomedical Engineering
- Nanotechnology
- Oncology
Background:
- Bladder cancer is a prevalent malignancy, necessitating advanced combination therapies.
- Gene and chemotherapeutic drug combinations offer synergistic potential for bladder cancer treatment.
- Carrier systems are crucial for co-delivering hydrophilic genes and hydrophobic drugs, avoiding damaging high-shear techniques.
Purpose of the Study:
- To develop a dual-payload carrier system for co-delivering genes and drugs to bladder cancer cells.
- To create a formulation free of sonication and extrusion, protecting nucleic acids.
- To achieve targeted delivery of therapeutic payloads to tumor sites.
Main Methods:
- A dual-payload carrier system was formulated using flow mixing to complex plasmid (EGFP-NLS) with cationic polymer (CD-PEI2.5kD).
- Polyplexes were coated with lipid membranes, forming lipid-coated polyplexes (LCPs).
- LCPs were targeted to bladder cancer cells using a bacterial adhesive peptide sequence (RWFV).
Main Results:
- The targeted LCP achieved 50% plasmid and 16% pyrene encapsulation efficiency.
- Targeted LCP demonstrated 4-fold higher transfection efficiency in bladder cancer cells compared to non-targeted LCP.
- Curcumin-loaded targeted LCP significantly reduced viable cancer cells (45% at 5 µg/mL).
Conclusions:
- The study highlights the specificity of the RWFV targeting peptide for bladder tumor cells.
- The developed carrier platform successfully delivers dual payloads (gene and drug) to bladder tumor cells.
- This targeted delivery system shows promise for effective bladder cancer combination therapy.
