Abstract
Purpose
Human pancreatic adenocarcinoma is a major leading cause of cancer mortality in the United States. Given that current strategies are relatively ineffective against this disease, new treatments are being developed. Liposomes possessing relatively high cationic lipid content preferentially accumulate in tumor angiogenic vessels compared to vessels in normal tissues. We therefore seek to develop cationic liposomes for targeting pancreatic tumor vessels.
Materials and Methods
We report development of 5-fluorouracil (5-FU) and doxorubicin hydrochloride (DOX) loaded in PEGylated cationic liposomes (PCLs). We evaluate cell association, intracellular fate, and cytotoxicity. Human pancreatic cancer cells HPAF-II and Capan-1, and endothelial cells HMEC-1 and HUVEC were used in this study. Intratumoral distribution of PCLs in (HPAF-II) tumors was determined by intravital microscopy.
Results
HUVEC and HMEC-1 were most susceptible to 5-FU after 24 and 48 h, compared to HPAF-II and Capan-1. We observed >90% incorporation of 5-FU and DOX in PCLs for 3–20 mol% preparations, with reduced incorporation for >20 mol% formulations. PCLs showed significantly higher association with human endothelial versus pancreatic cancer cells, and improved growth inhibitory properties of DOX. Intravital microscopy revealed distribution of PCLs along HPAF-II vessels.
Conclusions
Targeting human pancreatic cancer with PCLs may represent a rational alternative to conventional strategies.
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Abbreviations
- Capan-1 and HPAF-II:
-
human pancreatic cancer cells
- Chol:
-
cholesterol
- DOPC:
-
1,2-dioleoyl-sn-glycerol-phosphatidylcholine
- DOPE-PEG:
-
1,2-dioleoyl-sn-glycero-3-phosphoethanolamine-5000
- DOTAP:
-
1,2-dioleoyl-3-trimethylammonium-propane
- DOX:
-
doxorubicin hydrochloride
- EBM-2:
-
endothelial cell basal medium
- FITC-dextran:
-
fluorescein isothiocyanate-dextran
- HMEC-1:
-
human microvascular endothelial cells
- HUVEC:
-
human umbilical vein endothelial cell
- MEME:
-
eagle’s minimum essential medium
- PBS:
-
phosphate buffer saline
- PCLs:
-
PEGylated cationic liposomes
- Rhodamine-DPPE:
-
1,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine
- SCID:
-
severe combined immunodeficient
- 5-FU:
-
5-fluorouracil
References
B. F. El-Rayes, N. V. Adsay, and P. A. Philip. Pancreatic cancer: the evolving role of systemic therapy. Expert Opin. Pharmacother. 2:1939–1947 (2001).
V. Heinemann. Present and future treatment of pancreatic cancer. Semin. Oncol. 29:23–31 (2002).
L. Rosenberg. Pancreatic cancer: a review of emerging therapies. Drugs 1071–1089:1071–1089 (2000).
M. L. Rothenberg. New developments in chemotherapy for patients with advanced pancreatic cancer. Oncology 10:18–22 (1996).
D. P. Ryan and C. G. Willet. Management of locally advanced adenocarcinoma of the pancreas. Hematol./Oncol. Clin. North Am. 16:95–103 (2002).
H. G. Beger, B. Rau, F. Gansauge, B. Poch, and K. H. Link. Treatment of pancreatic cancer: challenge of the facts. World J. Surg. 27:1075–84 (2003).
R. Wilkowski, M. Thoma, C. Bruns, E. Duhmke, and V. Heinemann. Combined chemoradiotherapy for isolated local recurrence after primary resection of pancreatic cancer. JOP 11:34–40 (2006).
C. Morizane, T. Okusaka, Y. Ito, H. Ueno, M. Ikeda, Y. Takezako, Y. Kagami, and H. Ikeda. Chemoradiotherapy for locally advanced pancreatic carcinoma in elderly patients. Oncology 68:432–437 (2005).
M. Reni, S. Cordio, C. Milandri, P. Passoni, E. Bonetto, C. Oliani, G. Luppi, R. Nicoletti, L. Galli, R. Bordanaro, A. Passardi, A. Zerbi, G. Balzano, L. Aldrighetti, C. Straudacher, E. Villa, and V. Di Carlo. Gemcitabine versus cisplatin, epirubicin, fluorouracil, and gemcitabine in advanced pancreatic cancer: a randomised controlled multicentrre phase III trial. Lancet Oncol. 6:352–353 (2005).
D. A. Karlin, J. R. Stroehlein, R. W. Bennetts, R. D. Jones, L. J. Heifetz, and P. S. Mahal. Phase I–II study of the combination of 5-FU, doxorubicin, mitomycin, and semustine IFAMMe) in the treatment of adenocarcinoma of the stomach, gastroesophageal junction, and pancreas. Cancer Treat. Rep. 66:1613–1617 (1982).
D. J. Wagener, Q. van Hoesel, G. S. H. Yap, W. J. Hoogenraad, T. Wobbes, and S. P. Strijk. Phase II trial of 5-fluorouracil, adriamycin and cisplatin (FAP) followed by radiation and 5-fluorouracil in locally advanced pancreatic cancer. Cancer Chemother. Pharmacol. 25:131–134 (1989).
R. B. Campbell, D. Fukumura, E. B. Brown, M. L. Mazzola, Y. Izumi, R. K. Jain, V. P. Torchilin, and L. L. Munn. Cationic charge determines the distribution of liposomes between the vascular and extravascular compartments of tumors. Cancer Res. 62:6831–6836 (2002).
D. G. Hirst, J. Denekamp, and B. Hobson. Proliferation kinetics of endothelial and tumour cells in three mouse mammary carcinomas. Cell Tissue Kinet. 15:251–61 (1982).
J. Denekamp and B. Hobson. Endothelial-cell proliferation in experimental tumours. Br. J. Cancer 46:711–720 (1982).
J. Folkman. The role of angiogenesis in tumor growth. Semin. Cancer Biol. 3:65–71 (1992).
C. M. Lee, T. Tanaka, T. Murai, M. Kondo, J. Kimura, W. Su, T. Kitagawa, T. Ito, H. Matsuda, and M. Miyasaka. Novel chondroitin sulfate-binding cationic liposomes loaded with cisplatin efficiently suppress the local growth and liver metastasis of tumor cells in vivo. Cancer Res. 62:4282–4288 (2002).
R. Kunstfeld, G. Weckenhauser, U. Michaelis, M. Teifel, W. Umek, K. Naujoks, K. Wolff, and P. Petzelbauer. Paclitaxel encapsulated in cationic liposomes diminishes tumor angiogenesis and melanoma growth in a “humanized” SCID mouse model. J. Invest. Dermatol. 120:476–82 (2003).
S. Strieth, M. E. Eichhorn, B. Sauer, B. Schulze, M. Teifel, U. Michaelis, and M. Dellian. Neovascular targeting chemotherapy: encapsulation of paclitaxel in cationic liposome impairs functional tumor microvasculature. Int. J. Cancer 110:117–124 (2004).
S. Sengupta, P. Tyagi, S. Chandra, V. Kochupillai, and S. K. Gupta. Encapsulation in cationic liposomes enhances antitumour efficacy and reduces the toxicity of etoposide, a topo-isomerase II inhibitor. Pharmacology 62:163–171 (2001).
P. Skehan, R. Storeng, D. Scudiero, A. Monks, J. McMohan, J. T. Warren, H. Bokesch, S. Kenney, and M. R. Boyd. New colorimetric cytotoxicity assay for anticancer-drug screening. J. Natl. Cancer Inst. 82:1107–1112 (1990).
L. Ricotti, A. Tesei, F. D. Paola, P. Ulivi, G. L. Frassineti, C. Milandri, D. Amadori, and W. Zoli. In vitro schedule-dependent Interaction between Docetaxel and Gemcitabine in human gastric cancer cell lines. Clin. Cancer Res. 9:900–905 (2003).
F. Szoka and D. Papahadjopoulos. Comparative properties and methods of preparation of lipid vesicles (liposomes). Annu. Rev. Biophys. Bioeng. 9:467–508 (1980).
J. W. McLean, E. A. Fox, P. Baluk, P. B. Bolton, A. Haskell, R. Pearlman, G. Thurston, E. Y. Umemoto, and D. M. McDonald. Organ-specific endothelial cell uptake of cationic liposome-DNA complexes in mice. Am. J. Physiol. 273:387–404 (1997).
G. Thurston, J. W. McLean, M. Rizen, P. Baluk, A. Haskell, T. J. Murphy, D. Hanahan, and D. M. McDonald. Cationic liposomes target angiogenic endothelial cells in tumors and chronic inflammation in mice. J. Clin. Invest. 101:1401–1413 (1998).
G. Weckbecker. Biochemical pharmacology and analysis of fluoropyrimidines alone and in combination with modulators. Pharmacol. Ther. 50:367–424 (1991).
R. B. Campbell, S. V. Balasubramanian, and R. M. Straubinger. Physical properties of phospholipid-cationic lipid interactions: Influences on domain structure, liposome size and cellular uptake. Biochim. Biophys. Acta 1512:27–39 (2001).
R. B. Campbell, S. V. Balasubramanian, and R. M. Straubinger. Influence of cationic lipids on the stability and membrane properties of paclitaxel-containing liposomes. J. Pharm. Sci. 90: 1091–1105 (2001).
C. R. Dass. Cytotoxicity issues pertinent to lipoplex-mediated gene therapy in-vivo. J. Pharm. Pharmacol. 54:593–601 (2002).
J. L. Bramson, C. A. Bodner, and R. W. Graham. Activation of host antitumoral responses by cationic lipid/DNA complexes. Cancer Gen. Ther. 7:353–359 (2000).
M. C. Filion and N. C. Phillips. Anti-inflammatory activity of cationic lipids. Br. J. Pharmacol. 122:551–557 (1997).
D. Papahadjopoulos, T. Allen, A. Gabizon, E. Mayhew, K. Matthay, S. K. Huang, K. D. Lee, M. C. Woddle, D. D. Lasic, C. Redemann, and F. J. Martin. Sterically stabilized liposomes: improvements in pharmacokinetics and anti-tumor efficacy. Proc. Natl. Acad. Sci. USA 88:11460–11464 (1991).
T. M. Allen, C. Hansen, F. Martin, C. Redemann, and A. Yau-Young. Liposomes containing synthetic lipid derivatives of poly(ethyleneglycol) show prolonged circulation half-lives in vivo. Biochim. Biophys. Acta. 1066:29–36 (1991).
A. Gabizon, R. Catane, U. Beatrice, B. Kaufman, T. Safra, R. Cohen, A. H. Martin, and Y. Barenholz. Prolonged circulation time and enhanced accumulation in malignant exudates of doxorubicin encapsulated in polyethylene-glycol coated liposomes. Cancer Res. 54:987–992 (1994).
T. S. Levchenko, R. Rammohan, A. N. Lukyanov, K. R. Whiteman, and V. P. Torchilin. Liposome clearance in mice: the effect of a separate and combined presence of surface charge and polymer coating. Int. J. Pharm. 240:95–102 (2002).
J. Denekamp. Vasculature as a target for tumor therapy. Prog. Appl. Microcirc. 4:28–38 (1984).
L. C. Mounkes, W. Zhong, G. Cipres-Palacin, T. D. Heath, and R. Debs. Proteoglycans mediate cationic liposome-DNA complex-based gene delivery in vitro and in vivo. J. Biol. Chem. 1998:26164–26170 (1998).
K. A. Mislickand, and J. D. Baldeschwieler. Evidence for the role of proteoglycans in cation-mediated gene transfer. Proc. Natl. Acad. Sci. U. S. A. 93:12349–12354 (1996).
J. H. Levrat, C. Palevody, M. Daumas, G. Ratovo, and E. Hollande. Differentiation of the human pancreatic adenocarcinoma cell line (Capan-1) in culture and co-culture with fibroblasts dome formation. Int. J. Cancer 42:615–621 (1988).
B. Sipos, S. Moser, H. Kalthoff, V. Torok, M. Lohr, and G. Kloppel. A comprehensive characterization of pancreatic ductal carcinoma cell lines: towards the establishment of an in vitro research platform. Virchows Arch. 442:444–452 (2003).
A. Rahman, S. R. Husain, J. Siddiqui, M. Verma, M. Agresti, M. Center, A. R. Safa, and R. I. Glazer. Liposome-mediated modulation of multidrug resistance in human HL-60 leukemia cells. J. Natl. Cancer Inst. 84:1909–1915 (1992).
A. R. Thierry, A. Dritschilo, and A. Rahman. Effect of liposomes on P-glycoprotein function in multidrug resistant cells. Biochem. Biophys. Res. Commun. 187:1098–1105 (1992).
D. A. Gewirtz. A critical evaluation of the mechanisms of action proposed for the antitumor effects of anthracycline antibiotics adriamycin and daunorubicin. Biochem. Pharmacol. 57:724–741 (1999).
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Kalra, A.V., Campbell, R.B. Development of 5-FU and Doxorubicin-Loaded Cationic Liposomes against Human Pancreatic Cancer: Implications for Tumor Vascular Targeting. Pharm Res 23, 2809–2817 (2006). https://doi.org/10.1007/s11095-006-9113-3
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DOI: https://doi.org/10.1007/s11095-006-9113-3