Abstract
Effective gene therapy for disseminated metastatic cancer is currently impossible because of poor delivery of vector to target sites. Modification of viral vectors to target advanced cancer has long been a challenge. In this study, we aimed to redirect adenovirus tropism to infect prostate cancer cells via α6β1 integrins, whose expression is upregulated during prostate cancer progression. To ablate normal mechanisms of infection and provide a framework for attachment of targeting ligands, viruses were non-genetically modified with pHPMA-ONp polymer. Addition of polymer-coated virus to prostate cells showed significantly reduced transgene expression compared with unmodified virus. To restore infectivity, an α6-integrin binding peptide (-SIKVAV-) derived from laminin was incorporated onto the surface of the polymer-coated viruses. Photon correlation spectroscopic analysis revealed a small increase in the mean diameter of the particles following retargeting. Addition of -SIKVAV- peptide restored virus infectivity of PC-3 cells in a ligand concentration-dependent manner that was significantly improved following removal of unincorporated polymer and peptide. Competition assays using cells preincubated with Ad5 fiber protein or free -SIKVAV- peptide confirmed that entry of retargeted viruses was mediated via the incorporated ligand. Application of retargeted viruses to a panel of human cell lines revealed varying levels of transduction efficiency. Flow cytometric analysis of cells using anti-α6 integrin and anti-β1 integrin antibodies demonstrated that for prostate cells, greater transduction efficiency correlated with higher levels of expression of both integrin subunits. Furthermore with the exception of LNCaP cells, increased α6β1 integrin expression correlated with advanced disease. Intravenous administration of retargeted viruses to tumor-bearing mice resulted in slower plasma clearance and greatly reduced liver tropism, and hence toxicity compared with unmodified virus, while maintaining reporter gene expression in the tumor. The data suggest that YESIKVAVS-retargeted viruses have potential for systemic delivery for the treatment of metastatic disease.
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References
Rein DT, Breidenbach M, Wu HJ, Han T, Haviv YS, Wang MH et al. Gene transfer to cervical cancer with fiber-modified adenoviruses. Int J Cancer 2004; 111: 698–704.
Bauerschmitz GJ, Kanerva A, Wang MH, Herrmann I, Shaw DR, Strong TV et al. Evaluation of a selectively oncolytic adenovirus for local and systemic treatment of cervical cancer. Int J Cancer 2004; 111: 303–309.
Mercurio AM . Laminin receptors: achieving specificity through cooperation. Trends Cell Biol 1995; 5: 419–423.
Belkin AM, Stepp MA . Integrins as receptors for laminins. Microsc Res Tech 2000; 51: 280–301.
Stingl J, Eirew P, Ricketson I, Shackleton M, Vaillant F, Choi D et al. Purification and unique properties of mammary epithelial stem cells. Nature 2006; 439: 993–997.
Shackleton M, Vaillant F, Simpson KJ, Stingl J, Smyth GK, Asselin-Labat ML et al. Generation of a functional mammary gland from a single stem cell. Nature 2006; 439: 84–88.
Lin CS, Zhang K, Kramer R . Alpha-6 integrin is up-regulated in step increments accompanying neoplastic transformation and tumorigenic conversion of human fibroblasts. Cancer Res 1993; 53: 2950–2953.
Knox JD, Cress AE, Clark V, Manriquez L, Affinito KS, Dalkin BL et al. Differential expression of extracellular matrix molecules and the alpha 6-integrins in the normal and neoplastic prostate. Am J Pathol 1994; 145: 167–174.
Rabinovitz I, Nagle RB, Cress AE . Integrin alpha-6 expression in human prostate carcinoma-cells is associated with a migratory and invasive phenotype in-vitro and in-vivo. Clin Exp Metast 1995; 13: 481–491.
Vogelmann R, Kreuser ED, Adler G, Lutz MP . Integrin α6β1 role in metastatic behavior of human pancreatic carcinoma cells. Int J Cancer 1999; 80: 791–795.
Nejjari M, Hafdi Z, Dumortier J, Bringuier AF, Feldman G, Scoazec JY . 6β1 integrin expression in hepatocarcinoma cells: regulation and role in cell adhesion and migration. Int J Cancer 1999; 83: 518–525.
Takenaka K, Shibuya M, Takeda Y, Hibino S, Gemma A, Ono Y et al. Altered expression and function of beta 1 integrins in a highly metastatic human lung adenocarcinoma cell line. Int J Oncol 2000; 17: 1187–1194.
Liu AY . Differential expression of cell surface molecules in prostate cancer cells. Cancer Res 2000; 60: 3429–3434.
Bonaccorsi L, Carloni V, Muratori M, Salvadori A, Giannini A, Carini M et al. Androgen receptor expression in prostate carcinoma cells suppresses α6β4 integrin-mediated invasive phenotype. Endocrinology 2000; 141: 3172–3182.
Davis TL, Rabinovitz I, Futscher BW, Schnolzer M, Burger F, Liu YG et al. Identification of a novel structural variant of the α6 integrin. J Biol Chem 2001; 276: 26099–26106.
Edlund M, Miyamoto T, Sikes RA, Ogle R, Laurie GW, Farach-Carson MC et al. Integrin expression and usage by prostate cancer cell lines on laminin substrata. Cell Growth Differ 2001; 12: 99–107.
Bella-DeOcampo D, Kleinman HK, Webber MM . The role of α6β1 integrin and EGF in normal and malignant acinar morphogenesis of human prostatic epithelial cells. Mutat Res 2001; 480–481: 209–217.
Dyce OH, Ziober AF, Weber RS, Miyazaki K, Khariwala SS, Feldman M et al. Integrins in head and neck squamous cell carcinoma invasion. Laryngoscope 2002; 112: 2025–2032.
Shimizu H, Koyama N, Asada M, Yoshimatsu K . Aberrant expression of integrin and erbB subunits in breast cancer cell lines. Int J Oncol 2002; 21: 1073–1079.
Shaw LM, Chao C, Wewer UM, Mercurio AM . Function of the integrin alpha 6 beta 1 in metastatic breast carcinoma cells assessed by expression of a dominant-negative receptor. Cancer Res 1996; 56: 959–963.
Wewer UM, Shaw LM, Albrechtsen R, Mercurio AM . The integrin alpha 6 beta 1 promotes the survival of metastatic human breast carcinoma cells in mice. Am J Pathol 1997; 151: 1191–1198.
Nagle RB, Hao JS, Knox JD, Dalkin BL, Clark V, Cress AE . Expression of hemidesmosomal and extracellular matrix proteins by normal and malignant human prostate tissue. Am J Pathol 1995; 146: 1498–1507.
Tashiro K, Sephel GC, Weeks B, Sasaki M, Martin GR, Kleinman HK et al. A synthetic peptide containing the IKVAV sequence from the A chain of laminin mediates cell attachment, migration and neurite outgrowth. J Biol Chem 1989; 264: 16174–16182.
Faisal Khan KM, Falcone DJ . Role of laminin in matrix induction of macrophage urokinase-type plasminogen activator and 92-kDa metalloproteinase expression. J Biol Chem 1997; 272: 8270–8275.
Vliagoftis H, Metcalfe DD . Characterization of adhesive interactions between mast cells and laminin isoforms: evidence of a principle role for α6 integrin. Immunology 1997; 92: 553–560.
Fisher KD, Stallwood Y, Green NK, Ulbrich K, Mautner V, Seymour LW . Polymer-coated adenovirus permits efficient retargeting and evades neutralising antibodies. Gene Therapy 2001; 8: 341–348.
Cussenot O, Berthon P, Berger R, Mowszowicz I, Faille A, Hojman F et al. Immortalization of human adult normal prostate epithelial cells by liposomes containing large T-SV40 gene. J Urol 1991; 146: 881–886.
Cussenot O, Berthon P, Cochandpriollet S, Maitland NJ, Leduc A . Immunocytochemical comparison of cultured normal epithelial prostatic cells with prostatic tissue sections. Exp Cell Res 1994; 214: 83–92.
Berthon P, Cussenot O, Hopwood L, Leduc A, Maitland NJ . Functional expression of SV40 in normal human epithelial and fibroblastic cells-differentiation pattern of non-tumorigenic cell lines. Int J Oncol 1995; 6: 333–343.
Macintosh CA . An analysis of the role of human chromosome 8 in carcinoma of the prostate in vivo and in vitro. PhD Thesis, University of York. 1998.
Maitland NJ, Macintosh CA, Hall J, Sharrard M, Quinn G, Lang S . In vitro models to study cellular differentiation and function in human prostate cancers. Radiat Res 2001; 155: 133–142.
Murakami P, McCaman MT . Quantitation of adenovirus DNA and virus particles with the Pico green fluorescent dye. Anal Biochem 1999; 274: 283–288.
Haase AT, Mautner V, Pereira HG . The immunogenicity of adenovirus structural proteins. J Immunol 1972; 108: 483–485.
Alemany R, Suzuki K, Curiel DT . Blood clearance rates of adenovirus type 5 in mice. J Gen Virol 2000; 81: 2605–2609.
Green NK, Herbert CW, Hale SJ, Hale AB, Mautner V, Harkins R et al. Extended plasma circulation time and decreased toxicity of polymer-coated adenovirus. Gene Therapy 2004; 11: 1256–1263.
van Rooijen N, Kors N, Van der Ende M, Dijkstra CD . Depletion and repopulation of macrophages in spleen and liver of rat after intravenous treatment with liposome-encapsulated dichloromethylene diphosphonate. Cell Tissue Res 1990; 260: 215–222.
Kuzmin AI, Finegold MJ, Eisensmith RC . Macrophage depletion increases the safety, efficacy and persistence of adenovirus-mediated gene transfer in vivo. Gene Therapy 1997; 4: 309–316.
Wolff G, Worgall S, van Rooijen N, Song WR, Harvey BG, Crystal RG . Enhancement of in vivo adenovirus-mediated gene transfer and expression by prior depletion of tissue macrophages in the target organ. J Virol 1997; 71: 624–629.
Schiedner G, Hertel S, Johnston M, Dries V, van Rooijen N, Kochanek S . Selective depletion or blockade of Kupffer cells leads to enhanced and prolonged hepatic transgene expression using high-capacity Adenoviral vectors. Mol Ther 2003; 7: 35–43.
Hemler ME, Crouse C, Sonnenberg A . Association of the VLA-alpha-6 subunit with a novel protein – a possible alternative to the common VLA-beta-1 subunit on certain cell lines. J Biol Chem 1989; 264: 6529–6535.
Carloni V, Romanelli RG, Mercurio AM, Pinzani M, Laffi G, Cotrozzi G et al. Knockout of α6β1-integrin expression reverses the transformed phenotype of hepatocarcinoma cells. Gastroenterology 1998; 115: 433–442.
Belkin AM, Stepp MA . Integrins as receptors for laminins. Microsc Res Tech 2000; 51: 280–301.
Kibbey MC, Corcoran ML, Wahl LM, Kleinman HK . Laminin SIKVAV peptide-induced angiogenesis in vivo is potentiated by neutrophils. J Cell Physiol 1994; 160: 185–193.
Kanemoto T, Martin GR, Hamilton TC, Fridman R . Effects of synthetic peptides and protease inhibitors on the interaction of a human ovarian-cancer cell line (NIH-OVCAR-3) with a reconstituted basement-membrane (Matrigel). Invas Metast 1991; 11: 84–92.
Tashiro K, Sephel GC, Greatorex D, Sasaki M, Shirashi N, Martin GR et al. The RGD containing site of the mouse laminin A chain is active for cell attachment, spreading, migration and neurite outgrowth. J Cell Physiol 1991; 146: 451–459.
Grant DS, Kinsella JL, Fridman R, Auerbach R, Piasecki BA, Yamada Y et al. Interaction of endothelial-cells with a laminin-A chain peptide (SIKVAV) in vitro and induction of angiogenic behavior in vivo. J Cell Physiol 1992; 153: 614–625.
Kibbey MC, Grant DS, Kleinman HK . Role of the SIKVAV site of laminin in promotion of angiogenesis and tumor-growth – an in vivo matrigel model. J Natl Cancer Inst 1992; 84: 1633–1638.
Sweeney TM, Kibbey MC, Zain M, Fridman R, Kleinman HK . Basement-membrane and the SIKVAV laminin-derived peptide promote tumor-growth and metastasis. Cancer Metastasis Rev 1991; 10: 245–254.
Stack S, Gray RD, Pizzo SV . Modulation of plasminogen activation and type-IV collagenase activity by a synthetic peptide derived from the laminin A-chain. Biochemistry-US 1991; 30: 2073–2077.
Lang SH, Sharrard RM, Stark M, Villette JM, Maitland NJ . Prostate epithelial cell lines form spheroids with evidence of glandular differentiation in three-dimensional Matrigel cultures. Brit J Cancer 2001; 85: 590–599.
Lang SH, Hyde C, Reid IN, Hitchcock IS, Hart CA, Bryden AAG et al. Enhanced expression of vimentin in motile prostate cell lines and in poorly differentiated and metastatic prostate carcinoma. Prostate 2002; 52: 253–263.
Sonnenberg A, Linders CJT, Modderman PW, Damsky CH, Aumailley M, Timpl R . Integrin recognition of different cell-binding fragments of laminin (P1, E3, E8) and evidence that α6β1 but not α6β4 functions as a major receptor for fragment E8. J Cell Biol 1990; 110: 2145–2155.
Acknowledgements
We thank Sue Phipps and Oliver Hayward for assistance with virus production and Professor Norman Maitland for supplying the PNT1a, PNT-2 and P4E6 cell lines. We also thank Vivien Mautner for helpful advice and discussions. This work was funded by the European Union Framework 5 grant.
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Stevenson, M., Hale, A., Hale, S. et al. Incorporation of a laminin-derived peptide (SIKVAV) on polymer-modified adenovirus permits tumor-specific targeting via α6-integrins. Cancer Gene Ther 14, 335–345 (2007). https://doi.org/10.1038/sj.cgt.7701022
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DOI: https://doi.org/10.1038/sj.cgt.7701022
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