Therapeutic potential of capillary morphogenesis gene 2 extracellular vWA domain in tumour‑related angiogenesis

Ye,Lin; Sun,Ping-Hui; Sanders,Andrew  J.; Martin,Tracey  A.; Lane,Jane; Mason,Malcolm  D.; Jiang,Wen  G.

doi:10.3892/ijo.2014.2533

Therapeutic potential of capillary morphogenesis gene 2 extracellular vWA domain in tumour‑related angiogenesis

Authors:
- Lin Ye
- Ping-Hui Sun
- Andrew J. Sanders
- Tracey A. Martin
- Jane Lane
- Malcolm D. Mason
- Wen G. Jiang
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Affiliations: Metastasis and Angiogenesis Research Group, Cardiff University-Peking University Cancer Institute, Institute of Cancer and Genetics, Cardiff University School of Medicine, Cardiff, UK
Published online on: July 3, 2014 https://doi.org/10.3892/ijo.2014.2533
Pages: 1565-1573

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Abstract

Capillary morphogenesis gene 2 (CMG2) is a receptor of anthrax toxin and plays an important role in angiogenesis. It has been shown to be involved in the cell adhesion and motility of various cell types, including epithelia and endothelia. The present study aimed to examine the therapeutic potential of targeting CMG2 to prevent tumour‑related new vasculature. The full-length coding sequence of the human CMG2 gene and different fragments of the CMG2 vWA domain were amplified and constructed into a mammalian expression plasmid vector. The effect of CMG2 and its vWA domain on endothelial cells and angiogenesis was assessed using relevant in vitro, ex vivo and in vivo models. The overexpression of CMG2 enhanced the adhesion of endothelial cells to extracellular matrix, but was negatively associated with cell migration. Overexpression of CMG2 and the vWA domain fragments inhibited the tubule formation and migration of endothelial cells. Small peptides based on the amino acid sequence of the CMG2 vWA domain fragments potently inhibited in vitro tubule formation and ex vivo angiogenesis. One of the polypeptides, LG20, showed an inhibitory effect on in vivo tumour growth of cancer cells which were co-inoculated with the vascular endothelial cells. CMG2 is a potential target for treating tumour‑related angiogenesis. The polypeptides based on the CMG2 vWA domain can potently inhibit in vitro and ex vivo angiogenesis, which may contribute to the inhibitory effect on in vivo tumour growth. Further investigations are required to shed light on the machinery and may provide a novel therapeutic approach for inhibition of angiogenesis in cancer management.

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1	Bell SE, Mavila A, Salazar R, et al: Differential gene expression during capillary morphogenesis in 3D collagen matrices: regulated expression of genes involved in basement membrane matrix assembly, cell cycle progression, cellular differentiation and G-protein signaling. J Cell Sci. 114:2755–2773. 2001.
2	Scobie HM, Rainey GJ, Bradley KA and Young JA: Human capillary morphogenesis protein 2 functions as an anthrax toxin receptor. Proc Natl Acad Sci USA. 100:5170–5174. 2003. View Article : Google Scholar : PubMed/NCBI
3	Bradley KA, Mogridge J, Mourez M, Collier RJ and Young JA: Identification of the cellular receptor for anthrax toxin. Nature. 414:225–229. 2001. View Article : Google Scholar : PubMed/NCBI
4	Carson-Walter EB, Watkins DN, Nanda A, Vogelstein B, Kinzler KW and St Croix B: Cell surface tumor endothelial markers are conserved in mice and humans. Cancer Res. 61:6649–6655. 2001.PubMed/NCBI
5	Emsley J, King SL, Bergelson JM and Liddington RC: Crystal structure of the I domain from integrin alpha2beta1. J Biol Chem. 272:28512–28517. 1997. View Article : Google Scholar : PubMed/NCBI
6	Davies G, Cunnick GH, Mansel RE, Mason MD and Jiang WG: Levels of expression of endothelial markers specific to tumour-associated endothelial cells and their correlation with prognosis in patients with breast cancer. Clin Exp Metastasis. 21:31–37. 2004. View Article : Google Scholar
7	Rmali KA, Watkins G, Harrison G, Parr C, Puntis MC and Jiang WG: Tumour endothelial marker 8 (TEM-8) in human colon cancer and its association with tumour progression. Eur J Surg Oncol. 30:948–953. 2004. View Article : Google Scholar : PubMed/NCBI
8	Rmali KA, Puntis MC and Jiang WG: Prognostic values of tumor endothelial markers in patients with colorectal cancer. World J Gastroenterol. 11:1283–1286. 2005. View Article : Google Scholar : PubMed/NCBI
9	Davies G, Rmali KA, Watkins G, Mansel RE, Mason MD and Jiang WG: Elevated levels of tumour endothelial marker-8 in human breast cancer and its clinical significance. Int J Oncol. 29:1311–1317. 2006.PubMed/NCBI
10	Hanks S, Adams S, Douglas J, et al: Mutations in the gene encoding capillary morphogenesis protein 2 cause juvenile hyaline fibromatosis and infantile systemic hyalinosis. Am J Hum Genet. 73:791–800. 2003. View Article : Google Scholar : PubMed/NCBI
11	Dowling O, Difeo A, Ramirez MC, et al: Mutations in capillary morphogenesis gene-2 result in the allelic disorders juvenile hyaline fibromatosis and infantile systemic hyalinosis. Am J Hum Genet. 73:957–966. 2003. View Article : Google Scholar : PubMed/NCBI
12	Tumer L, Kasapkara C, Fong K, Serdaroglu A and McGrath JA: Hyaline fibromatosis syndrome resulting from a new homozygous missense mutation, p. Gly116Val, in ANTXR2. J Dermatol. 40:677–678. 2013. View Article : Google Scholar : PubMed/NCBI
13	Al Sinani S, Al Murshedy F and Abdwani R: Infantile systemic hyalinosis: a case report with a novel mutation. Oman Med J. 28:53–55. 2013.PubMed/NCBI
14	Wang YY, Wen CQ, Wei Z and Jin X: A novel splice site mutation in ANTXR2 (CMG2) gene results in systemic hyalinosis. J Pediatr Hematol Oncol. 33:e355–e357. 2011. View Article : Google Scholar : PubMed/NCBI
15	Yan SE, Lemmin T, Salvi S, et al: In-depth analysis of hyaline fibromatosis syndrome frameshift mutations at the same site reveal the necessity of personalized therapy. Hum Mutat. 34:1005–1017. 2013. View Article : Google Scholar : PubMed/NCBI
16	Martchenko M, Candille SI, Tang H and Cohen SN: Human genetic variation altering anthrax toxin sensitivity. Proc Natl Acad Sci USA. 109:2972–2977. 2012. View Article : Google Scholar : PubMed/NCBI
17	Reveille JD, Sims AM, Danoy P, et al: Genome-wide association study of ankylosing spondylitis identifies non-MHC susceptibility loci. Nat Genet. 42:123–127. 2010. View Article : Google Scholar : PubMed/NCBI
18	Guo C, Xia Y, Yang Q, Qiu R, Zhao H and Liu Q: Association of the ANTXR2 gene polymorphism and ankylosing spondylitis in Chinese Han. Scand J Rheumatol. 41:29–32. 2012. View Article : Google Scholar : PubMed/NCBI
19	Chen C, Zhang X and Wang Y: ANTXR2 and IL-1R2 polymorphisms are not associated with ankylosing spondylitis in Chinese Han population. Rheumatol Int. 32:15–19. 2012. View Article : Google Scholar : PubMed/NCBI
20	Peters DE, Zhang Y, Molinolo AA, et al: Capillary morphogenesis protein-2 is required for mouse parturition by maintaining uterine collagen homeostasis. Biochem Bioph Res Co. 422:393–397. 2012. View Article : Google Scholar : PubMed/NCBI
21	Reeves CV, Wang X, Charles-Horvath PC, et al: Anthrax toxin receptor 2 functions in ECM homeostasis of the murine reproductive tract and promotes MMP activity. PLoS One. 7:e348622012. View Article : Google Scholar : PubMed/NCBI
22	Reeves CV, Dufraine J, Young JA and Kitajewski J: Anthrax toxin receptor 2 is expressed in murine and tumor vasculature and functions in endothelial proliferation and morphogenesis. Oncogene. 29:789–801. 2010. View Article : Google Scholar : PubMed/NCBI
23	Bonnekoh B, Wevers A, Jugert F, Merk H and Mahrle G: Colorimetric growth assay for epidermal cell cultures by their crystal violet binding capacity. Arch Dermatol Res. 281:487–490. 1989. View Article : Google Scholar : PubMed/NCBI
24	Jiang WG, Davies G, Martin TA, et al: Targeting matrilysin and its impact on tumor growth in vivo: the potential implications in breast cancer therapy. Clin Cancer Res. 11:6012–6019. 2005. View Article : Google Scholar : PubMed/NCBI
25	Jiang WG, Hiscox S, Hallett MB, Scott C, Horrobin DF and Puntis MC: Inhibition of hepatocyte growth factor-induced motility and in vitro invasion of human colon cancer cells by gamma-linolenic acid. Br J Cancer. 71:744–752. 1995. View Article : Google Scholar : PubMed/NCBI
26	Rosen EM, Meromsky L, Setter E, Vinter DW and Goldberg ID: Smooth muscle-derived factor stimulates mobility of human tumor cells. Invasion Metastasis. 10:49–64. 1990.PubMed/NCBI
27	Jiang WG, Hiscox S, Singhrao SK, Nakamura T, Puntis MC and Hallett MB: Inhibition of HGF/SF-induced membrane ruffling and cell motility by transient elevation of cytosolic free Ca²⁺. Exp Cell Res. 220:424–433. 1995. View Article : Google Scholar : PubMed/NCBI
28	Jiang WG, Martin TA, Lewis-Russell JM, Douglas-Jones A, Ye L and Mansel RE: Eplin-alpha expression in human breast cancer, the impact on cellular migration and clinical outcome. Mol Cancer. 7:712008. View Article : Google Scholar : PubMed/NCBI
29	Cai J, Jiang WG and Mansel RE: Inhibition of the expression of VE-cadherin/catenin complex by gamma linolenic acid in human vascular endothelial cells, and its impact on angiogenesis. Biochem Biophys Res Commun. 258:113–118. 1999. View Article : Google Scholar : PubMed/NCBI
30	Grant DS, Tashiro K, Segui-Real B, Yamada Y, Martin GR and Kleinman HK: Two different laminin domains mediate the differentiation of human endothelial cells into capillary-like structures in vitro. Cell. 58:933–943. 1989. View Article : Google Scholar : PubMed/NCBI
31	Cai J, Jiang WG and Mansel RE: Inhibition of angiogenic factor- and tumour-induced angiogenesis by gamma linolenic acid. Prostaglandins Leukot Essent Fatty Acids. 60:21–29. 1999. View Article : Google Scholar : PubMed/NCBI
32	Jiang WG and Harding KG: Enhancement of wound tissue expansion and angiogenesis by matrix-embedded fibroblast (dermagraft), a role of hepatocyte growth factor/scatter factor. Int J Mol Med. 2:203–210. 1998.PubMed/NCBI
33	Chaudhary A and St Croix B: Selective blockade of tumor angiogenesis. Cell Cycle. 11:2253–2259. 2012. View Article : Google Scholar
34	Liu S, Zhang Y, Hoover B and Leppla SH: The receptors that mediate the direct lethality of anthrax toxin. Toxins (Basel). 5:1–8. 2013. View Article : Google Scholar : PubMed/NCBI
35	Pilpa RM, Bayrhuber M, Marlett JM, Riek R and Young JA: A receptor-based switch that regulates anthrax toxin pore formation. PLoS Pathog. 7:e10023542011. View Article : Google Scholar : PubMed/NCBI
36	Rogers MS, Christensen KA, Birsner AE, et al: Mutant anthrax toxin B moiety (protective antigen) inhibits angiogenesis and tumor growth. Cancer Res. 67:9980–9985. 2007. View Article : Google Scholar : PubMed/NCBI
37	Rmali KA, Puntis MC and Jiang WG: TEM-8 and tubule formation in endothelial cells, its potential role of its vW/TM domains. Biochem Biophys Res Commun. 334:231–238. 2005. View Article : Google Scholar : PubMed/NCBI
38	Abrami L, Leppla SH and van der Goot FG: Receptor palmitoylation and ubiquitination regulate anthrax toxin endocytosis. J Cell Biol. 172:309–320. 2006. View Article : Google Scholar : PubMed/NCBI
39	Liu S, Leung HJ and Leppla SH: Characterization of the interaction between anthrax toxin and its cellular receptors. Cell Microbiol. 9:977–987. 2007. View Article : Google Scholar : PubMed/NCBI
40	Garlick KM, Batty S and Mogridge J: Binding of filamentous actin to anthrax toxin receptor 1 decreases its association with protective antigen. Biochemistry. 51:1249–1256. 2012. View Article : Google Scholar : PubMed/NCBI
41	Cai C, Che J, Xu L, et al: Tumor endothelium marker-8 based decoys exhibit superiority over capillary morphogenesis protein-2 based decoys as anthrax toxin inhibitors. PLoS One. 6:e206462011. View Article : Google Scholar : PubMed/NCBI
42	Cryan LM, Bazinet L, Habeshian KA, et al: 1,2,3,4,6-Penta-O-galloyl-beta-D-glucopyranose inhibits angiogenesis via inhibition of capillary morphogenesis gene 2. J Med Chem. 56:1940–1945. 2013. View Article : Google Scholar : PubMed/NCBI
43	Cao S, Cryan L, Habeshian KA, et al: Phenolic compounds as antiangiogenic CMG2 inhibitors from Costa Rican endophytic fungi. Bioorg Med Chem Lett. 22:5885–5888. 2012. View Article : Google Scholar : PubMed/NCBI
44	Rogers MS, Cryan LM, Habeshian KA, et al: A FRET-based high throughput screening assay to identify inhibitors of anthrax protective antigen binding to capillary morphogenesis gene 2 protein. PLoS One. 7:e399112012. View Article : Google Scholar : PubMed/NCBI