Abstract
Basic fibroblast growth factor (FGF-2) is expressed in vascular endothelium during tumor neovascularization and angioproliferative diseases, including vascular tumors and Kaposi's sarcoma (KS). We have investigated the in vivo biological consequences of endothelial cell activation by endogenous FGF-2 in a mouse aortic endothelial cell line transfected with a retroviral expression vector harboring a human FGF-2 cDNA and the neomycin resistance gene. FGF-2 transfectants, named pZipbFGF2-MAE cells, caused the rapid growth of highly vascularized, non-infiltrating tumors when injected in nude mice. In contrast, lesions grew poorly when cells were injected in immunocompetent syngeneic animals. Histologically, the tumors had the appearance of hemangioendothelioma with spindled areas resembling KS and with numerous CD31+ blood vessels and lacunae. Southern blot analysis of tumor DNA, as well as disaggregation of the lesion followed by in vitro cell culture, revealed that less than 10% of the cells in the tumor mass retain FGF-2 overexpression and neomycin resistance at 6–8 weeks post-injection. Nevertheless, in vitro G418 selection allowed the isolation from the tumor of a FGF-2-overexpressing cell population showing biochemical and biological characteristics similar to those of pZipbFGF2-MAE cells, including the capacity to originate vascular lesions when re-injected in nude mice. To evaluate the effect of angiostatic compounds on the growth and vascularization of pZipbFGF2-MAE cell-induced lesions, nude mice were treated weekly (100mg/kg, i.p.) with the angiostatic sulfonated distamycin A derivative 2,2′-(carbonyl-bis-[imino-N-methyl-4,2-pyrrole carbonyl-imino-{N-methyl-4,2-pyrrole}carbonylimino])-bis-(1,5-naphthalene) disulfonic acid (PNU 153429). The results demonstrate that PNU 153429 inhibits the growth of the lesions and causes a ∼50% decrease in CD31+ microvessel density. In conclusion, the data indicate that FGF-2-overexpressing endothelial cells cause vascular lesions in immunodeficient mice which may represent a novel model for opportunistic vascular tumors suitable for the evaluation of angiostatic compounds.
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References
Enzinger FM, Weiss SW. Soft Tissue Tumors. St Louis, MO: Mosby-Year Book 1995: 579–677.
Folkman J. Clinical applications of research on angiogenesis. New Engl J Med 1995; 333, 1757–1763.
Levine AM. AIDS-related malignancies: the emerging epidemic. J Natl Cancer Inst 1993; 85, 1382–1387.
Sturzl M, Brandstetter H, Roth WK. Kaposi's sarcoma: a review of gene expression and ultrastructure of KS spindle cells in vivo. AIDS Res Human Retroviruses 1992; 8, 1753–1763.
Basilico C, Moscatelli D. The FGF family of growth factors and oncogenes. Adv Cancer Res 1992; 59, 115–165.
Florkiewicz RZ, Sommer A. Human basic fibroblast growth factor gene encodes four polypeptides: three initiate translation from non-AUG codons. Proc Natl Acad Sci USA 1989; 86, 3978–3981.
Gualandris A, Urbinati C, Rusnati M, Ziche M, Presta M. Interaction of high molecular weight basic fibroblast growth factor (bFGF) with endothelium: biological activity and intracellular fate of human recombinant Mr 24,000 bFGF. J Cell Physiol 1994; 161, 149–159.
Sato Y, Rifkin DB. Autocrine activities of basic fibroblast growth factor: regulation of endothelial cell movement, plasminogen activator synthesis, and DNA synthesis. J Cell Biol 1988; 107, 1199–1205.
Itoh H, Mukoyama M, Pratt RE, Dzau VJ. Specific blockade of basic fibroblast growth factor gene expression in endothelial cells by antisense oligonucleotide. Biochem Biophys Res Commun 1992; 188, 1205–1213.
Pepper MS, Sappino AP, Stocklin R, Montesano R, Orci L, Vassalli JD. Upregulation of urokinase receptor expression on migrating endothelial cells. J Cell Biol 1993; 122, 673–684.
Schulze-Osthoff K, Risau W, Vollmer E, Sorg C. In situ detection of basic fibroblast growth factor by highly specific antibodies. Am J Pathol 1990; 137, 85–92.
Takahashi JA, Mori H, Fukumoto M, et al. Gene expression of fibroblast growth factors in human gliomas and meningiomas: demonstration of cellular source of basic fibroblast growth factor mRNA and peptide in tumor tissues. Proc Natl Acad Sci USA 1990; 87, 5710–5714.
Ohtani H, Nakamura S, Watanabe Y, Mizoi T, Saku T, Nagura H. Immunocytochemical localization of basic fibroblast growth factor in carcinomas and inflammatory lesions of the human digestive tract. Lab Invest 1993; 68, 520–527.
Zagzag D, Miller DC, Sato Y, Rifkin DB, Burstein DE. Immunohistochemical localization of basic fibroblast growth factor in astrocytomas. Cancer Res 1990; 50, 7393–7398.
Statuto M, Ennas MG, Zamboni G, et al. Basic fibroblast growth factor in human pheochromo cytoma: a biochemical and immunohistochemical study. Int J Cancer 1993; 53, 5–10.
Peverali FA, Mandriota SJ, Ciana P, et al. Tumor cells secrete an angiogenic factor that stimulates basic fibroblast growth factor and urokinase expression in vascular endothelial cells. J Cell Physiol 1994; 161, 1–14.
Weich H, Iberg N, Klagsbrun M, Folkman J. Transcriptional regulation of basic fibroblast growth factor gene expression in capillary endothelial cells. J Cell Biochem 1991; 47, 158–194.
Cozzolino F, Torcia M, Lucibello M, et al. Cytokinemediated control of endothelial cell growth: interferon-α and interleukin-2 synergistically enhance basic fibroblast growth factor synthesis and induce release promoting cell growth in vitro and in vivo. J Clin Invest 1993; 91, 2504–2512.
Barillari G, Buonaguro L, Fiorelli V, et al. Effects of cytokines from activated immune cells on vascular cell growth and HIV-1 gene expression. J Immunol 1992; 149, 3727–3734.
Fiorelli V, Gendelman R, Samaniego F, Markham PD, Ensoli B. Cytokines from activated T cells induce normal endothelial cells to acquire the phenotypic and functional features of AIDS-Kaposi's sarcoma spindle cells. J Clin Invest 1995; 95, 1723–1734.
Albini A, Fontanini G, Masiello L, et al. Angiogenic potential in vivo by Kaposi sarcoma cell-free supernatants and HIV1-tat product: inhibition of KS-like lesions by TIMP-2. AIDS 1994; 8, 1237–1244.
Ensoli B, Gendelman R, Markham P, et al. Synergy between basic fibroblast growth factor and HIV-1 Tat protein in induction of Kaposi's sarcoma. Nature 1994; 371, 674–680.
Ensoli B, Markham P, Kao V, et al. Block of AIDS-Kaposi's sarcoma cell growth, angiogenesis, and lesion formation in nude mice by antisense oligonucleotide targeting basic fibroblast growth factor. A novel strategy for the therapy of KS. J Clin Invest 1994; 94, 1736–1746.
Takahashi K, Mulliken JB, Kozakewich HPW, Rogers RA, Folkman J, Ezekowitz RAB. Cellular markers that distinguish the phases of hemangioma during infancy and childhood. J Clin Invest 1994; 93, 2357–2364.
Gualandris A, Rusnati M, Belleri M, et al. Basic fibroblast growth factor overexpression in endothelial cells: an autocrine mechanism for angiogenesis and angioproliferative diseases. Cell Growth Different 1996; 7, 147–160.
Sola F, Farao M, Ciomei M, Pastori A, Mongelli N, Grandi M. FCE 27266, a sulfonic distamycin derivative, inhibits experimental and spontaneous lung and liver metastasis. Invasion Metastasis 1995; 15, 222–231.
Vecchi A, Garlanda C, Lampugnani MG, et al. Monoclonal antibodies specific for endothelial cells of mouse blood vessels. Their application in the identification of adult and embryonic endothelium. Eur J Cell Biol 1994; 63, 247–254.
Sola F, Farao M, Pesenti E, et al. Antitumor activity of FCE 26644 a new growth-factor complexing molecule. Cancer Chemother Pharmacol 1995; 36, 217–222.
Molinari Tosatti MP, Parolini S, Rosa D, et al. Basement membrane components in normal, dysplastic and neoplastic epithelia of the uterine cervix. An immunohistochemical study on the distribution of laminin, type IV collagen and fibronectin. Cervix 1991; 9, 177–181.
Pepper MS, Montesano R, Vassalli JD, Orci L. Chondrocytes inhibit endothelial sprout formation in vitro: evidence for involvement of a transforming growth factor-beta. J Cell Physiol 1991; 146, 170–179.
Kleinman HK, McGarvey ML, Hassell JR, et al. Basement membrane complexes with biological activity. Biochemistry 1986; 25, 312–318.
Ciomei M, Pastori W, Mariani M, Sola F, Grandi M, Mongelli N. New sulfonated distamycin A derivatives with bFGF complexing activity. Biochem Pharmacol 1994; 47, 295–302.
Bastaki M, Nelli EE, Dell'Era P, et al. Basic fibroblast growth factor-induced angiogenic phenotype in mouse endothelium. A study of aortic and microvascular endothelial cell lines. Arterioscler Thromb Vasc Biol 1997; 17, 454–464.
Salahuddin SZ, Nakamura S, Biberfeld P, et al. Angiogenic properties of Kaposi's sarcoma-derived cells after long-term culture in vitro. Science 1988; 242, 430–433.
Williams RL, Risau W, Zerwes HG, Drexler H, Aguzzi A, Wagner EF. Endothelioma cells expressing the polyoma middle T oncogene induce hemangiomas by host cell recruitment. Cell 1898; 57, 1053–1063.
Garlanda C, Parravicini C, Sironi M, et al. Progressive growth in immunodeficient mice and host cell recruitment by mouse endothelial cells transformed by polyoma middle-sized T antigen: implications for the pathogenesis of opportunistic vascular tumors. Proc Natl Acad Sci USA 1994; 91, 7291–7295.
Ensoli B, Nakamura S, Salahuddin SZ, et al. AIDS-Kaposi's sarcoma-derived cells express cytokines with autocrine and paracrine growth effects. Science 1989; 243, 223–226.
Taraboletti G, Belotti D, Dejana E, et al. Endothelial cell migration and invasiveness are induced by a soluble factor produced by murine endothelioma cells transformed by polyoma virus middle T oncogene. Cancer Res 1993; 53, 3812–3816.
Bussolino F, De Rossi M, Sica A, et al. Murine endothelioma cell lines transformed by polyoma middle T oncogene as target for and producers of cytokines. J Immunol 1991; 147, 2122–2129.
Jouanneau J, Moens G, Bourgeois Y, Poupon MF, Thiery JP. A minority of carcinoma cells producing acidic fibroblast growth factor induces a community effect for tumor progression. Proc Natl Acad Sci USA 1994; 91, 286–290.
Fan TP, Jaggar R, Bicknell R. Controlling the vasculature: angiogenesis, anti-angiogenesis and vascular targeting of gene therapy. Trends Pharmacol Sci 1995; 16, 57–66.
Sola F, Farao M, Marsiglio A, Mariani M, Grandi, M. Inhibition of lung and liver tumor colonies in mice pretreated with suramin. Invasion Metastasis 1993; 13, 163–168.
Pesenti E, Sola F, Mongelli N, et al. Suramin prevents neovascularization and tumor growth through blocking of basic fibroblast growth factor activity. Br J Cancer 1992; 66, 367–372.
Fan TP. Angiosuppressive therapy for cancer. Trends Pharmacol Sci 1994; 15, 33–36.
Hawkins MJ. Clinical trials of antiangiogenic agents. Curr Opin Oncol 1995; 7, 90–93.
Braddock PS, Hu DE, Fan TP, Stratford IJ, Harris AL, Bicknell R. A structure-activity analysis of antagonism of the growth factor and angiogenic activity of basic fibroblast growth factor by suramin and related polyanions. Br J Cancer 1994; 69, 890–898.
Takano S, Gately S, Neville ME, et al. Suramin, an anticancer and angiosuppressive agent, inhibits endothelial cell binding of basic fibroblast growth factor, migration, proliferation, and induction of urokinase-type plasminogen activator. Cancer Res 1994; 54, 2654–2660.
Sola F, Biasoli G, Pesenti E, et al. In vivo activity of novel sulphonic derivatives of distamycin A. In: Steiner R, Weisz PB, Langer R (eds), Angiogenesis, Key Principles-Science-Technology-Medicine. Basel: Birkhauser Verlag 1992: 459–462.
Taraboletti G, Garofalo A, Belotti D, et al. Inhibition of angiogenesis and murine hemangioma growth by batimastat, a synthetic inhibitor of matrix metalloproteinases. J Natl Cancer Inst 1995; 87, 293–298.
Ezekowitz RAB, Mulliken JB, Folkman J. Interferon alfa-2a therapy for life-threatening hemangiomas of infancy. New Engl J Med 1992; 326, 1456–1463.
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Sola, F., Gualandris, A., Belleri, M. et al. Endothelial cells overexpressing basic fibroblast growth factor (FGF-2) induce vascular tumors in immunodeficient mice. Angiogenesis 1, 102–116 (1997). https://doi.org/10.1023/A:1018309200629
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DOI: https://doi.org/10.1023/A:1018309200629