Targeting diphtheria toxin A-chain transcription to activated endothelial cells using an E-selectin promoter Article DOI:
Cite this article as: Maxwell, I.H., Kaletta, C., Naujoks, K. et al. Angiogenesis (2003) 6: 31. doi:10.1023/A:1025894616613 Abstract
Targeting the transcription of a toxin gene to activated endothelial cells might be used for inhibiting angiogenesis in solid tumors. As a model, we transiently transfected human endothelial cells (HUVEC) in culture with expression plasmids for the toxic A-chain of diphtheria toxin (DT-A), using electroporation (achieving ≈70% transfection efficiency). Protein synthesis in HUVEC was highly sensitive to DT-A expression from constitutive viral promoters. E-selectin is strongly expressed on HUVEC activated by TNF
α or TPA. We therefore tested a human E-selectin promoter (−547 to +33) for targeting transcription of DT-A or reporter genes to HUVEC. Luciferase reporters were efficiently expressed in HUVEC from this promoter, with or without an enhancer responsive to Ets-1. Expression was increased by TNF α or TPA. DT-A showed highly preferential expression (increased by TNF α or TPA) in HUVEC, compared with WI38 human fibroblasts. HUVEC expressing DT-A were killed via apoptosis. Overall expression levels were influenced by alternative ‘backbone’ sequences used in the expression plasmids. We propose that delivery of transcriptionally regulated expression plasmids for DT-A in vivo, using cationic lipids that show preferential accumulation in activated or proliferating endothelium, may offer a novel means of inhibiting undesired angiogenesis. diphtheria toxin endothelial cells E-selectin promoter transcription References
Folkman J. Incipient angiogenesis. J Natl Cancer Inst 2000; 92: 94–5.
Novak K. Angiogenesis inhibitors revised and revived at AACR. Nat Med 2002; 8: 427.
Deplanque G, Harris AL. Anti-angiogenic agents: Clinical trial design and therapies in development. Eur J Cancer 2000; 36: 1713–24.
Pappenheimer AMJ. Diphtheria toxin. Ann Rev Biochem 1977; 46: 69–94.
Falnes PO, Ariansen S, Sandvig K, Olsnes S. Requirement for prolonged action in the cytosol for optimal protein synthesis inhibition by diphtheria toxin. J Biol Chem 2000; 275: 4363–68.
Kochi SK, Collier RJ. DNA fragmentation and cytolysis in U937 cells treated with diphtheria toxin or other inhibitors of protein synthesis. Exp Cell Res 1993; 208: 296–302.
Kageyama A, Kusano I, Tamura T et al. Comparison of the apoptosis-inducing abilities of various protein synthesis inhibitors in U937 cells. Biosci Biotechnol Biochem 2002; 66: 835–9.
Maxwell IH, Maxwell F, Glode LM. Regulated expression of a diphtheria toxin A-chain gene transfected into human cells: Possible strategy for inducing cancer cell suicide. Cancer Res 1986; 46: 4660–4.
Massuda ES, Dunphy EJ, Redman RA et al. Regulated expression of the diphtheria toxin A chain by a tumor-specific chimeric transcription factor results in selective toxicity for alveolar rhabdomyosarcoma cells. Proc Natl Acad Sci USA 1997; 94: 14701–6.
Palmiter RD, Behringer RR, Quaife CJ et al. Cell lineage ablation in transgenic mice by cell-specific expression of a toxin gene. Cell 1987; 50: 435–43.
Breitman ML, Clapoff S, Rossant J et al. Genetic ablation: Targeted expression of a toxin gene causes microphthalmia in transgenic mice. Science 1987; 238: 1563–5.
Burrows HL, Birkmeier TS, Seasholtz AF, Camper SA. Targeted ablation of cells in the pituitary primordia of transgenic mice. Mol Endocrinol 1996; 10: 1467–77.
Ying S, Jansen HT, Lehman MN et al. Retinal degeneration in cone photoreceptor cell-ablated transgenic mice. Mol Vis 2000; 24: 101–8.
Thurston G, McLean JW, Rizen M et al. Cationic liposomes target angiogenic endothelial cells in tumors and chronic inflammation in mice. J Clin Invest 1998; 101: 1401–13.
Augustin HG, Kozian DH, Johnson RC. Differentiation of endothelial cells: Analysis of the constitutive and activated endothelial cell phenotypes. Bioessays 1994; 16: 901–6.
Brooks PC, Clark RA, Cheresh DA. Requirement of vascular integrin alpha v beta 3 for angiogenesis. Science 1994; 264: 569–71.
Harari OA, Wickham TJ, Stocker CJ et al. Targeting an adenoviral gene vector to cytokine-activated vascular endothelium via Eselectin. Gene Ther 1999; 6: 801–7.
Simon SI, Hu Y, Vestweber D, Smith CW. Neutrophil tethering on E-selectin activates beta 2 integrin binding to ICAM-1 through a mitogen-activated protein kinase signal transduction pathway. J Immunol 2000; 164: 4348–58.
Fox SB, Turner GD, Gatter KC, Harris AL. The increased expression of adhesion molecules ICAM-3, E-and P-selectins on breast cancer endothelium. J Pathol 1995; 177: 369–76.
Ye C, Kiriyama K, Mistuoka C et al. Expression of E-selectin on endothelial cells of small veins in human colorectal cancer. Int J Cancer 1995; 61: 455–60.
Walton T, Wang JL, Ribas A et al. Endothelium-specific expression of an E-selectin promoter recombinant adenoviral vector. Anticancer Res 1998; 18: 1357–60.
Murakami T, Mataki C, Nagao C et al. The gene expression profile of human umbilical vein endothelial cells stimulated by tumor necrosis factor alpha using DNA microarray analysis. J Atheroscler Thromb 2000; 7: 39–44.
Whitley MZ, Thanos D, Read MA et al. Striking similarity in the organization of the E-selectin and beta interferon gene promoters. Mol Cell Biol 1994; 14: 6464–75.
Maxwell IH, Harrison GS, Wood WM, Maxwell F. A DNA cassette containing a trimerized SV40 polyadenylation signal which efficiently blocks spurious plasmid-initiated transcription. Biotechniques 1989; 7: 276–80.
Day RN, Kawecki M, Berry D. Dual-function reporter protein for analysis of gene expression in living cells. Biotechniques 1998; 25: 848–56.
Schlaeger TM, Bartunkova S, Lawitts JA et al. Uniform vascularendothelial-cell-specific gene expression in both embryonic and adult transgenic mice. Proc Natl Acad Sci USA 1997; 94: 3058–63.
Maxwell IH, Maxwell F. Electroporation of mammalian cells with a firefly luciferase expression plasmid: Kinetics of transient expression differ markedly among cell types. DNA 1988; 7: 557–62.
Duke RC, Cohen JJ. Quantitation of apoptotic index and cell viability using fluorescent dyes. Curr Protocols Immunol 1992; Suppl 3. 3.17.1.
Cotran RS, Gimbrone MA, Bevilacqua MP et al. Induction and detection of a human endothelial activation antigen
. J Exp Med 1986; 164: 661–6.
Wang N, Chintala SK, Fini ME, Schuman JS. Activation of a tissue-specific stress response in the aqueous outflow pathway of the eye defines the glaucoma disease phenotype. Nat Med 2001; 7: 304–9.
Wernert N, Raes MB, Lassalle P et al. c-ets1 proto-oncogene is a transcription factor expressed in endothelial cells during tumor vascularization and other forms of angiogenesis in humans. Am J Pathol 1992; 140: 119–27.
Li H, Oehrlein SA, Wallerath T et al. Activation of protein kinase C alpha and/or epsilon enhances transcription of the human endothelial nitric oxide synthase gene. Mol Pharmacol 1998; 53: 630–7.
Kunstfeld R, Wickenhauser G, Michaelis U et al. Paclitaxel encapsulated in cationic liposomes diminishes tumor angiogenesis and melanoma growth in a ‘humanized’ SCID mouse model. J Invest Dermatol 2003; 120: 476–82.
McLean JW, Fox EA, Baluk P et al. Organ-specific endothelial cell uptake of cationic liposome-DNA complexes in mice. Am J Physiol 1997; 273: H387–404.
Arap W, Pasqualini R, Ruoslahti E. Chemotherapy targeted to tumor vasculature. Curr Opin Oncol 1998; 10: 560–5.
Maxwell F, Maxwell IH, Glode LM. Cloning, sequence determination, and expression in transfected cells of the coding sequence for the tox176 attenuated diphtheria toxin A chain. Mol Cell Biol 1987; 7: 1576–79.
Hood JD, Bednarski M, Frausto R et al. Tumor regression by targeted gene delivery to the neovasculature. Science 2002; 296: 2404–7.
Google Scholar Copyright information
© Kluwer Academic Publishers 2003