Skip to main content

The NG2/HMP Proteoglycan as a Cancer Therapeutic Target

  • Protocol
Target Discovery and Validation Reviews and Protocols

Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 361))

  • 1388 Accesses

Abstract

Neuroepithelial cells of the central nervous system constitute neuroglia (astrocytes, oligodendrocytes, and microglia), ependyma, and neurons, which make up the stromal cells of the brain. The stromal tissue organization of the brain is tightly regulated, but occasionally the signals that define the normal contexts become disrupted and result in cancer. Malignant progression is then maintained by cross-talks between the tumor and its stroma, where the activated stroma nurtures the proliferative and invasive neoplastic cells, by providing neovasculature, extracelluar matrix components, and stimulatory growth factors. The NG2/HMP plays a major role in tumor-stroma activation through alterations in cellular adhesion, migration, proliferation, and vascular morphogenesis. Therapeutic strategies specifically targeting NG2/HMP may be useful in normalizing the tumor stroma and may reduce the toxic side effects when used in combination with conventional treatments.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 84.99
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 169.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 109.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

Similar content being viewed by others

References

  1. Rutka, J. T., Apodaca, G., Stern, R., and Rosenblum, M. (1988) The extracellular matrix of the central and peripheral nervous systems: structure and function. J. Neurosurg. 69, 155–170.

    CAS  PubMed  Google Scholar 

  2. Tillet, E., Ruggiero, F., Nishiyama, A., and Stallcup, W. B. (1997) The membrane-spanning proteoglycan NG2 binds to collagens V and VI through the central nonglobular domain of its core protein. J. Biol. Chem. 272, 10,769–10,776.

    CAS  PubMed  Google Scholar 

  3. Burg, M. A., Nishiyama, A., and Stallcup, W. B. (1997) A central segment of the NG2 proteoglycan is critical for the ability of glioma cells to bind and migrate toward type VI collagen. Exp. Cell Res., 235, 254–264.

    CAS  PubMed  Google Scholar 

  4. Schlessinger, J., Lax, I., and Lemmon, M. (1995) Regulation of growth factor activation by proteoglycans: what is the role of the low affinity receptors? Cell, 83, 357–360.

    CAS  PubMed  Google Scholar 

  5. David, G. (1993) Integral membrane heparan sulfate proteoglycans. FASEB J. 7, 1023–1030.

    CAS  PubMed  Google Scholar 

  6. Stallcup, W. B. (2002) The NG2 proteoglycan: past insights and future prospects. J. Neurocytol. 31, 423–435.

    CAS  PubMed  Google Scholar 

  7. Stallcup, W. B., and Dahlin-Huppe, K. (2001) Chondroitin sulfate and cytoplasmic domain-dependent membrane targeting of the NG2 proteoglycan promotes retraction fiber formation and cell polarization. J. Cell Sci. 114, 2315–2325.

    CAS  PubMed  Google Scholar 

  8. Nishiyama, A., Lin, X. H., and Stallcup, W. B. (1995) Generation of truncated forms of the NG2 proteoglycan by cell surface proteolysis. Mol. Biol. Cell 6, 1819–1832.

    CAS  PubMed  Google Scholar 

  9. Yamaguchi, Y. (2000) Chondroitin sulfate proteoglycans in the nervous system. Marcel Dekker, Inc., New York.

    Google Scholar 

  10. Yamaguchi, Y. (2001) Heparin sulfate proteoglycans in the nervous system: Their diverse roles in neurogenesis,axon guidance, and synaptogenesis. Sem. Cell Dev. Biol. 12, 96–106.

    Google Scholar 

  11. Songyang, Z., Fanning, A. S., Fu, C., et al. (1997) Recognition of unique carboxyl-terminal motifs by distinct PDZ domains. Science 275, 73–77.

    CAS  PubMed  Google Scholar 

  12. Barritt, D. S., Pearn, M. T., Zisch, A. H., et al. (2000) The multi-PDZ domain protein MUPP1 is a cytoplasmic ligand for the membrane-spanning proteoglycan NG2. J. Cell Biochem. 79, 213–224.

    CAS  PubMed  Google Scholar 

  13. Nishiyama, A., Dahlin, K. J., Prince, J. T., Johnstone, S. R., and Stallcup, W. B. (1991) The primary structure of NG2, a novel membrane-spanning proteoglycan. J. Cell Biol. 114, 359–371.

    CAS  PubMed  Google Scholar 

  14. Yu, H., Chen, J. K., Feng, S., Dalgarno, D. C., Brauer, A. W., and Schreiber, S. L. (1994) Structural basis for the binding of proline-rich peptides to SH3 domains. Cell 76, 933–945.

    CAS  PubMed  Google Scholar 

  15. Stallcup, W. B., Beasley, L. and Levine, J. (1983) Cell-surface molecules that characterize different stages in the development of cerebellar interneurons. Cold Spring Harb Symp. Quant. Biol. 48Pt 2, 761–774.

    CAS  PubMed  Google Scholar 

  16. Houghton, A. N., Eisinger, M., Albino, A. P., Cairncross, J. G., and Old, L. J. (1982) Surface antigens of melanocytes and melanomas. Markers of melanocyte differentiation and melanoma subsets. J. Exp. Med., 156, 1755–1766.

    CAS  PubMed  Google Scholar 

  17. Bumol, T. F., Walker, L. E., and Reisfeld, R. A. (1984) Biosynthetic studies of proteoglycans in human melanoma cells with a monoclonal antibody to a core glycoprotein of chondroitin sulfate proteoglycans. J. Biol. Chem., 259, 12,733–12,741.

    CAS  PubMed  Google Scholar 

  18. Bumol, T. F. and Reisfeld, R. A. (1982) Unique glycoprotein-proteoglycan complex defined by monoclonal antibody on human melanoma cells. Proc. Natl. Acad. Sci. USA 79, 1245–1249.

    CAS  PubMed  Google Scholar 

  19. Schneider, S., Bosse, F., D’Urso, D., et al. (2001) The AN2 protein is a novel marker for the Schwann cell lineage expressed by immature and nonmyelinating Schwann cells. J. Neurosci., 21, 920–933.

    Google Scholar 

  20. Niehaus, A., Stegmuller, J., Diers-Fenger, M., and Trotter, J. (1999) Cell-surface glycoprotein of oligodendrocyte progenitors involved in migration. J. Neurosci. 19, 4948–4961.

    CAS  PubMed  Google Scholar 

  21. Trapp, B. D., Nishiyama, A., Cheng, D., and Macklin, W. (1997) Differentiation and death of premyelinating oligodendrocytes in developing rodent brain. J. Cell. Biol. 137, 459–468.

    CAS  PubMed  Google Scholar 

  22. Reynolds, R. and Hardy, R. (1997) Oligodendroglial progenitors labeled with the O4 antibody persist in the adult rat cerebral cortex in vivo. J. Neurosci. Res. 47, 455–470.

    CAS  PubMed  Google Scholar 

  23. Nishiyama, A., Lin, X. H., Giese, N., Heldin, C. H., and Stallcup, W. B. (1996) Interaction between NG2 proteoglycan and PDGF alpha-receptor on O2A progenitor cells is required for optimal response to PDGF. J. Neurosci. Res., 43, 315–330.

    CAS  PubMed  Google Scholar 

  24. Nishiyama, A., Lin, X. H., Giese, N., Heldin, C. H., and Stallcup, W. B. (1996) Co-localization of NG2 proteoglycan and PDGF alpha-receptor on O2A progenitor cells in the developing rat brain. J. Neurosci. Res., 43, 299–314.

    CAS  PubMed  Google Scholar 

  25. Levine, J. M., Stincone, F., and Lee, Y. S. (1993) Development and differentiation of glial precursor cells in the rat cerebellum. Glia, 7, 307–321.

    CAS  PubMed  Google Scholar 

  26. Keirstead, H. S., Levine, J. M., and Blakemore, W. F. (1998) Response of the oligodendrocyte progenitor cell population (defined by NG2 labelling) to demyelination of the adult spinal cord. Glia 22, 161–170.

    CAS  PubMed  Google Scholar 

  27. Luskin, M. B., Parnavelas, J. G., and Barfield, J. A. (1993) Neurons, astrocytes, and oligodendrocytes of the rat cerebral cortex originate from separate progenitor cells: an ultrastructural analysis of clonally related cells. J. Neurosci. 13, 1730–1750.

    CAS  PubMed  Google Scholar 

  28. Levison, S. W., Young, G. M., and Goldman, J. E. (1999) Cycling cells in the adult rat neocortex preferentially generate oligodendroglia. J. Neurosci. Res. 57, 435–446.

    CAS  PubMed  Google Scholar 

  29. Levison, S. W. and Goldman, J. E. (1993) Both oligodendrocytes and astrocytes develop from progenitors in the subventricular zone of postnatal rat forebrain. Neuron 10, 201–212.

    CAS  PubMed  Google Scholar 

  30. Grove, E. A., Williams, B. P., Li, D. Q., Hajihosseini, M., Friedrich, A. and Price, J. (1993) Multiple restricted lineages in the embryonic rat cerebral cortex. Development 117, 553–561.

    CAS  PubMed  Google Scholar 

  31. Richardson, W. D., Pringle, N., Mosley, M. J., Westermark, B., and Dubois-Dalcq, M. (1988) A role for platelet-derived growth factor in normal gliogenesis in the central nervous system. Cell, 53, 309–319.

    CAS  PubMed  Google Scholar 

  32. Raff, M. C., Lillien, L. E., Richardson, W. D., Burne, J. F., and Noble, M. D. (1988) Platelet-derived growth factor from astrocytes drives the clock that times oligodendrocyte development in culture. Nature 333, 562–565.

    CAS  PubMed  Google Scholar 

  33. Ellison, J. A. and de Vellis, J. (1994) Platelet-derived growth factor receptor is expressed by cells in the early oligodendrocyte lineage. J. Neurosci. Res. 37, 116–128.

    CAS  PubMed  Google Scholar 

  34. Tanaka, K., Nogawa, S., Ito, D., et al. (2001) Activation of NG2-positive oligodendrocyte progenitor cells during post-ischemic reperfusion in the rat brain. Neuroreport 12, 2169–2174.

    CAS  PubMed  Google Scholar 

  35. Levine, J. M., Reynolds, R., and Fawcett, J. W. (2001) The oligodendrocyte precursor cell in health and disease. Trends Neurosci. 24, 39–47.

    CAS  PubMed  Google Scholar 

  36. Shee, W. L., Ong, W. Y., and Lim, T. M. (1998) Distribution of perlecan in mouse hippocampus following intracerebroventricular kainate injections. Brain Res. 799, 292–300.

    CAS  PubMed  Google Scholar 

  37. Reynolds, R., Cenci di Bello, I., Dawson, M., and Levine, J. (2001) The response of adult oligodendrocyte progenitors to demyelination in EAE. Prog. Brain Res. 132, 165–174.

    Google Scholar 

  38. McTigue, D. M., Wei, P., and Stokes, B. T. (2001) Proliferation of NG2-positive cells and altered oligodendrocyte numbers in the contused rat spinal cord. J. Neurosci. 21, 3392–3400.

    CAS  PubMed  Google Scholar 

  39. Levine, J. M., Enquist, L. W., and Card, J. P. (1998) Reactions of oligodendrocyte precursor cells to alpha herpesvirus infection of the central nervous system. Glia 23, 316–328.

    CAS  PubMed  Google Scholar 

  40. Levine, J. M. (1994) Increased expression of the NG2 chondroitin-sulfate proteoglycan after brain injury. J. Neurosci., 14, 4716–4730.

    CAS  PubMed  Google Scholar 

  41. Back, S. A., Luo, N. L., Borenstein, N. S., Levine, J. M., Volpe, J. J., and Kinney, H. C. (2001) Late oligodendrocyte progenitors coincide with the developmental window of vulnerability for human perinatal white matter injury. J. Neurosci. 21, 1302–1312.

    CAS  PubMed  Google Scholar 

  42. Jones, L. L., Yamaguchi, Y., Stallcup, W. B., and Tuszynski, M. H. (2002) NG2 is a major chondroitin sulfate proteoglycan produced after spinal cord injury and is expressed by macrophages and oligodendrocyte progenitors. J. Neurosci. 22, 2792–2803.

    CAS  PubMed  Google Scholar 

  43. Humphries, M. J. (2000) Integrin cell adhesion receptors and the concept of agonism. Trends Pharmacol. Sci. 21, 29–32.

    CAS  PubMed  Google Scholar 

  44. Garrigues, H. J., Lark, M. W., Lara, S., Hellstrom, I., Hellstrom, K. E., and Wight, T. N. (1986) The melanoma proteoglycan: restricted expression on microspikes, a specific microdomain of the cell surface. J. Cell Biol. 103, 1699–1710.

    CAS  PubMed  Google Scholar 

  45. Harper, J. R., Bumol, T. F., and Reisfeld, R. A. (1984) Characterization of monoclonal antibody 155.8 and partial characterization of its proteoglycan antigen on human melanoma cells. J. Immunol. 132, 2096–2104.

    CAS  PubMed  Google Scholar 

  46. de Vries, J. E., Keizer, G. D., te Velde, A. A., et al. (1986) Characterization of melanoma-associated surface antigens involved in the adhesion and motility of human melanoma cells. Int. J. Cancer 38, 465–473.

    PubMed  Google Scholar 

  47. Fukushi, J., Makagiansar, I. T., and Stallcup, W. B. (2004) NG2 proteoglycan promotes endothelial cell motility and angiogenesis via engagement of galectin-3 and alpha3beta1 integrin. Mol. Biol. Cell 15, 3580–3590.

    CAS  PubMed  Google Scholar 

  48. Gladson, C. L. (1999) The extracellular matrix of gliomas: modulation of cell function. J. Neuropathol. Exp. Neurol. 58, 1029–1040.

    CAS  PubMed  Google Scholar 

  49. Burg, M. A., Grako, K. A., and Stallcup, W. B. (1998) Expression of the NG2 proteoglycan enhances the growth and metastatic properties of melanoma cells. J. Cell Physiol. 177, 299–312.

    CAS  PubMed  Google Scholar 

  50. East, J. A., Mitchell, S. D., and Hart, I. R. (1993) Expression and function of the CD44 glycoprotein in melanoma cell lines. Melanoma Res. 3, 341–346.

    CAS  PubMed  Google Scholar 

  51. Kuppner, M. C., Van Meir, E., Gauthier, T., Hamou, M. F., and de Tribolet, N. (1992) Differential expression of the CD44 molecule in human brain tumours. Int., J. Cancer 50, 572–577.

    CAS  Google Scholar 

  52. Merzak, A., Koocheckpour, S., and Pilkington, G. J. (1994) CD44 mediates human glioma cell adhesion and invasion in vitro. Cancer Res. 54, 3988–3992.

    CAS  PubMed  Google Scholar 

  53. Akiyama, Y., Jung, S., Salhia, B., et al. (2001) Hyaluronate receptors mediating glioma cell migration and proliferation. J. Neurooncol. 53, 115–127.

    CAS  PubMed  Google Scholar 

  54. Midwood, K. S. and Salter, D. M. (2001) NG2/HMPG modulation of human articular chondrocyte adhesion to type VI collagen is lost in osteoarthritis. J. Pathol. 195, 631–635.

    CAS  PubMed  Google Scholar 

  55. Iida, J., Meijne, A. M., Spiro, R. C., Roos, E., Furcht, L. T., and McCarthy, J. B. (1995) Spreading and focal contact formation of human melanoma cells in response to the stimulation of both melanoma-associated proteoglycan (NG2) and alpha 4 beta 1 integrin. Cancer Res. 55, 2177–2185.

    CAS  PubMed  Google Scholar 

  56. Doane, K. J., Howell, S. J., and Birk, D. E. (1998) Identification and functional characterization of two type VI collagen receptors, alpha 3 beta 1 integrin and NG2, during avian corneal stromal development. Invest. Ophthalmol. Vis. Sci. 39, 263–275.

    CAS  PubMed  Google Scholar 

  57. Burg, M. A., Tillet, E., Timpl, R., and Stallcup, W. B. (1996) Binding of the NG2 proteoglycan to type VI collagen and other extracellular matrix molecules. J. Biol. Chem. 271, 26,110–26,116.

    CAS  PubMed  Google Scholar 

  58. Nishiyama, A. and Stallcup, W. B. (1993) Expression of NG2 proteoglycan causes retention of type VI collagen on the cell surface. Mol. Biol. Cell 4, 1097–1108.

    CAS  PubMed  Google Scholar 

  59. Stallcup, W. B., Dahlin, K., and Healy, P. (1990) Interaction of the NG2 chondroitin sulfate proteoglycan with type VI collagen. J. Cell Biol., 111, 3177–3188.

    CAS  PubMed  Google Scholar 

  60. Kuo, H. J., Maslen, C. L., Keene, D. R., and Glanville, R. W. (1997) Type VI collagen anchors endothelial basement membranes by interacting with type IV collagen. J. Biol. Chem. 272, 26,522–26,529.

    CAS  PubMed  Google Scholar 

  61. Rand, J. H., Wu, X. X., Potter, B. J., Uson, R. R., and Gordon, R. E. (1993) Colocalization of von Willebrand factor and type VI collagen in human vascular subendothelium. Am. J. Pathol. 142, 843–850.

    CAS  PubMed  Google Scholar 

  62. Chintala, S. K., Sawaya, R., Gokaslan, Z. L., Fuller, G., and Rao, J. S. (1996) Immunohistochemical localization of extracellular matrix proteins in human glioma, both in vivo and in vitro. Cancer Lett. 101, 107–114.

    CAS  PubMed  Google Scholar 

  63. Paulus, W., Roggendorf, W., and Schuppan, D. (1988) Immunohistochemical investigation of collagen subtypes in human glioblastomas. Virchows Arch. A Pathol. Anat. Histopathol. 413, 325–332.

    CAS  PubMed  Google Scholar 

  64. Chekenya, M., Rooprai, H. K., Davies, D., Levine, J. M., Butt, A. M., and Pilkington, G. J. (1999) The NG2 chondroitin sulfate proteoglycan: role in malignant progression of human brain tumours. Int J. Dev. Neurosci. 17, 421–435.

    CAS  PubMed  Google Scholar 

  65. Chekenya, M., Enger, P., Thorsen, F., et al. (2002) The glial precursor proteoglycan, NG2, is expressed on tumour neovasculature by vascular pericytes in human malignant brain tumours. Neuropathol. Appl. Neurobiol. 28, 367–380.

    CAS  PubMed  Google Scholar 

  66. Lemmon, V., Farr, K. L., and Lagenaur, C. (1989) L1-mediated axon outgrowth occurs via a homophilic binding mechanism. Neuron 2, 1597–1603.

    CAS  PubMed  Google Scholar 

  67. Edgar, D., Timpl, R., and Thoenen, H. (1984) The heparin-binding domain of laminin is responsible for its effects on neurite outgrowth and neuronal survival. EMBO J. 3, 1463–1468.

    CAS  PubMed  Google Scholar 

  68. Fidler, P. S., Schuette, K., Asher, R. A., et al. (1999) Comparing astrocytic cell lines that are inhibitory or permissive for axon growth: the major axon-inhibitory proteoglycan is NG2. J. Neurosci. 19, 8778–8788.

    CAS  PubMed  Google Scholar 

  69. Fawcett, J. W., and Asher, R. A. (1999) The glial scar and central nervous system repair. Brain Res. Bull 49, 377–391.

    CAS  PubMed  Google Scholar 

  70. Dou, C. L., and Levine, J. M. (1994) Inhibition of neurite growth by the NG2 chondroitin sulfate proteoglycan. J. Neurosci. 14, 7616–7628.

    CAS  PubMed  Google Scholar 

  71. Pedersen, P. H., Marienhagen, K., Mork, S., and Bjerkvig, R. (1993) Migratory pattern of fetal rat brain cells and human glioma cells in the adult rat brain. Cancer Res. 53, 5158–5165.

    CAS  PubMed  Google Scholar 

  72. Giordana, M. T., Germano, I., Giaccone, G., Mauro, A., Migheli, A., and Schiffer, D. (1985) The distribution of laminin in human brain tumors: an immunohistochemical study. Acta. NeuroPathol. (Berl) 67, 51–57.

    CAS  Google Scholar 

  73. Noble, M.-P. (2001) Glial Restricted Precursors. Humana Press, Totowa, New Jersey.

    Google Scholar 

  74. Sugimoto, Y., Taniguchi, M., Yagi, T., Akagi, Y., Nojyo, Y., and Tamamaki, N. (2001) Guidance of glial precursor cell migration by secreted cues in the developing optic nerve. Development 128, 3321–3330.

    CAS  PubMed  Google Scholar 

  75. Fang, X., Burg, M. A., Barritt, D., Dahlin-Huppe, K., Nishiyama, A., and Stallcup, W. B. (1999) Cytoskeletal reorganization induced by engagement of the NG2 proteoglycan leads to cell spreading and migration. Mol. Biol. Cell 10, 3373–3387.

    CAS  PubMed  Google Scholar 

  76. Lin, X. H., Dahlin-Huppe, K., and Stallcup, W. B. (1996) Interaction of the NG2 proteoglycan with the actin cytoskeleton. J. Cell Biochem. 63, 463–477.

    CAS  PubMed  Google Scholar 

  77. Bogler, O., Wren, D., Barnett, S. C., Land, H., and Noble, M. (1990) Cooperation between two growth factors promotes extended self-renewal and inhibits differentiation of oligodendrocyte-type-2 astrocyte (O-2A) progenitor cells. Proc. Natl. Acad. Sci. USA 87, 6368–6372.

    CAS  PubMed  Google Scholar 

  78. Baron, W., Metz, B., Bansal, R., Hoekstra, D., and de Vries, H. (2000) PDGF and FGF-2 signaling in oligodendrocyte progenitor cells: regulation of proliferation and differentiation by multiple intracellular signaling pathways. Mol. Cell. Neurosci. 15, 314–329.

    CAS  PubMed  Google Scholar 

  79. Redwine, J. M., Blinder, K. L., and Armstrong, R. C. (1997) In situ expression of fibroblast growth factor receptors by oligodendrocyte progenitors and oligodendrocytes in adult mouse central nervous system. J. Neurosci. Res. 50, 229–237.

    CAS  PubMed  Google Scholar 

  80. Grako, K. A., and Stallcup, W. B. (1995) Participation of the NG2 proteoglycan in rat aortic smooth muscle cell responses to platelet-derived growth factor. Exp. Cell Res. 221, 231–240.

    CAS  PubMed  Google Scholar 

  81. Goretzki, L., Burg, M. A., Grako, K. A., and Stallcup, W. B. (1999) High-affinity binding of basic fibroblast growth factor and platelet-derived growth factor-AA to the core protein of the NG2 proteoglycan. J. Biol. Chem. 274, 16,831–16,837.

    CAS  PubMed  Google Scholar 

  82. Brekke, C., Lundervold, A., Enger, P. Ø., et al. (2006) NG2 expression regulates vascular morphology and function in human brain tumours. NeuroImage 29, 965–976. Epub Oct 25, 2005.

    CAS  PubMed  Google Scholar 

  83. Chekenya, M., Hjelstuen, M., Enger, P. O., et al. (2002) NG2 proteoglycan promotes angiogenesis-dependent tumor growth in CNS by sequestering angiostatin. FASEB J. 16, 586–588.

    CAS  PubMed  Google Scholar 

  84. Hermanson, M., Funa, K., Koopmann, J., et al. (1996) Association of loss of heterozygosity on chromosome 17p with high platelet-derived growth factor alpha receptor expression in human malignant gliomas. Cancer Res. 56, 164–171.

    CAS  PubMed  Google Scholar 

  85. Nister, M., Libermann, T. A., Betsholtz, C., et al. (1988) Expression of messenger RNAs for platelet-derived growth factor and transforming growth factor-alpha and their receptors in human malignant glioma cell lines. Cancer Res. 48, 3910–3918.

    CAS  PubMed  Google Scholar 

  86. Fleming, T. P., Saxena, A., Clark, W. C., et al. (1992) Amplification and/or overexpression of platelet-derived growth factor receptors and epidermal growth factor receptor in human glial tumors. Cancer Res. 52, 4550–4553.

    CAS  PubMed  Google Scholar 

  87. Bergers, G., and Benjamin, L. E. (2003) Tumorigenesis and the angiogenic switch. Nat. Rev. Cancer 3, 401–410.

    CAS  PubMed  Google Scholar 

  88. Ozerdem, U., Monosov, E., and Stallcup, W. B. (2002) NG2 Proteoglycan Expression by Pericytes in Pathological Microvasculature. Microvasc. Res. 63, 129–134.

    CAS  PubMed  Google Scholar 

  89. Ozerdem, U., Grako, K. A., Dahlin-Huppe, K., Monosov, E., and Stallcup, W. B. (2001) NG2 proteoglycan is expressed exclusively by mural cells during vascular morphogenesis. Dev. Dyn 222, 218–227.

    CAS  PubMed  Google Scholar 

  90. Miller, B., Sheppard, A. M., Bicknese, A. R., and Pearlman, A. L. (1995) Chondroitin sulfate proteoglycans in the developing cerebral cortex: the distribution of neurocan distinguishes forming afferent and efferent axonal pathways. J. Comp. Neurol. 355, 615–628.

    CAS  PubMed  Google Scholar 

  91. Lindahl, P., Johansson, B. R., Leveen, P., and Betsholtz, C. (1997) Pericyte loss and microaneurysm formation in PDGF-B-deficient mice. Science 277, 242–245.

    CAS  PubMed  Google Scholar 

  92. Hellstrom, M., Kalen, M., Lindahl, P., Abramsson, A., and Betsholtz, C. (1999) Role of PDGF-B and PDGFR-beta in recruitment of vascular smooth muscle cells and pericytes during embryonic blood vessel formation in the mouse. Development 126, 3047–3055.

    CAS  PubMed  Google Scholar 

  93. Murfee, W. L., Skalak, T. C., and Peirce, S. M. (2005) Differential arterial/venous expression of NG2 proteoglycan in perivascular cells along microvessels: identifying a venule-specific phenotype. Microcirculation 12, 151–160.

    CAS  PubMed  Google Scholar 

  94. Schwartz, S. M., Heimark, R. L., and Majesky, M. W. (1990) Developmental mechanisms underlying pathology of arteries. Physiol. Rev. 70, 1177–1209.

    CAS  PubMed  Google Scholar 

  95. Ross. (1993) The pathogenesis of atherosclerosis: a perspective for the 1990s. Nature 362, 801–809.

    CAS  PubMed  Google Scholar 

  96. Grako, K. A., Ochiya, T., Barritt, D., Nishiyama, A., and Stallcup, W. B. (1999) PDGF (alpha)-receptor is unresponsive to PDGF-AA in aortic smooth muscle cells from the NG2 knockout mouse. J. Cell Sci. 112(Pt 6), 905–915.

    CAS  PubMed  Google Scholar 

  97. Orlidge, A., and D’Amore, P. A. (1987) Inhibition of capillary endothelial cell growth by pericytes and smooth muscle cells. J. Cell Biol. 105, 1455–1462.

    CAS  PubMed  Google Scholar 

  98. Asher, R. A., Morgenstern, D. A., Properzi, F., Nishiyama, A., Levine, J. M., and Fawcett, J. W. (2005) Two separate metalloproteinase activities are responsible for the shedding and processing of the NG2 proteoglycan in vitro. Mol. Cell Neurosci. 29, 82–96.

    CAS  PubMed  Google Scholar 

  99. Goretzki, L., Lombardo, C. R., and Stallcup, W. B. (2000) Binding of the NG2 proteoglycan to kringle domains modulates the functional properties of angiostatin and plasmin(ogen). J. Biol. Chem. 275, 28,625–28,633.

    CAS  PubMed  Google Scholar 

  100. Ashley, D. M., Batra, S. K., and Bigner, D. D. (1997) Monoclonal antibodies to growth factors and growth factor receptors: their diagnostic and therapeutic potential in brain tumors. J. Neurooncol. 35, 259–273.

    CAS  PubMed  Google Scholar 

  101. Harper, J. R., and Reisfeld, R. A. (1983) Inhibition of anchorage-independent growth of human melanoma cells by a monoclonal antibody to a chondroitin sulfate proteoglycan. J. Natl. Cancer Inst. 71, 259–263.

    CAS  PubMed  Google Scholar 

  102. Bumol, T. F., Wang, Q. C., Reisfeld, R. A., and Kaplan, N. O. (1983) Monoclonal antibody and an antibody-toxin conjugate to a cell surface proteoglycan of melanoma cells suppress in vivo tumor growth. Proc. Natl. Acad. Sci. USA 80, 529–533.

    CAS  PubMed  Google Scholar 

  103. Folkman, J. (1995) Angiogenesis in cancer, vascular, rheumatoid and other disease. Nat. Med. 1, 27–31.

    CAS  PubMed  Google Scholar 

  104. Burrows, F. J., and Thorpe, P. E. (1993) Eradication of large solid tumors in mice with an immunotoxin directed against tumor vasculature. Proc. Natl. Acad. Sci. USA 90, 8996–9000.

    CAS  PubMed  Google Scholar 

  105. Kerbel, R. S. (1997) A cancer therapy resistant to resistance. Nature 390, 335–336.

    CAS  PubMed  Google Scholar 

  106. Kerbel, R. S. (1991) Inhibition of tumor angiogenesis as a strategy to circumvent acquired resistance to anti-cancer therapeutic agents. Bioessays 13, 31–36.

    CAS  PubMed  Google Scholar 

  107. Boehm, T., Folkman, J., Browder, T., and O’Reilly, M. S. (1997) Antiangiogenic therapy of experimental cancer does not induce acquired drug resistance. Nature 390, 404–407.

    CAS  PubMed  Google Scholar 

  108. Natali, P. G., Imai, K., Wilson, B. S., et al. (1981) Structural properties and tissue distribution of the antigen recognized by the monoclonal antibody 653.40S to human melanoma cells. J. Natl. Cancer Inst. 67, 591–601.

    CAS  PubMed  Google Scholar 

  109. Ferrone, S. G. P., Natali, P. G., et al. (1983) A human high molecular weightmelanoma associated antigen (HMWW_MAA) defined by monoclonal antibodies: a useful marker to radioimage tumor lesions in patients with melanoma. Brookhaven National Laboratories. Associated Universities, Inc., Brookhaven, New York.

    Google Scholar 

  110. Giacomini, P., Natali, P., and Ferrone, S. (1985) Analysis of the interaction between a human high molecular weight melanoma-associated antigen and the monoclonal antibodies to three distinct antigenic determinants. J. Immunol. 135, 696–702.

    CAS  PubMed  Google Scholar 

  111. Lindmo, T., Boven, E., Cuttitta, F., Fedorko, J., and Bunn, P. A., Jr. (1984) Determination of the immunoreactive fraction of radiolabeled monoclonal antibodies by linear extrapolation to binding at infinite antigen excess. J. Immunol. Methods 72, 77–89.

    CAS  PubMed  Google Scholar 

  112. Kohler, G., and Milstein, C. (1975) Continuous cultures of fused cells secreting antibody of predefined specificity. Nature 256, 495–497.

    CAS  PubMed  Google Scholar 

  113. Winter, G., Griffiths, A. D., Hawkins, R. E., and Hoogenboom, H. R. (1994) Making antibodies by phage display technology. Annu. Rev. Immunol. 12, 433–455.

    CAS  PubMed  Google Scholar 

  114. Schaffitzel, C., Hanes, J., Jermutus, L., and Pluckthun, A. (1999) Ribosome display: an in vitro method for selection and evolution of antibodies from libraries. J. Immunol. Methods 231, 119–135.

    CAS  PubMed  Google Scholar 

  115. Giovannoni, L., Viti, F., Zardi, L., and Neri, D. (2001) Isolation of anti-angiogenesis antibodies from a large combinatorial repertoire by colony filter screening. Nucleic Acids Res. 29, E27.

    CAS  PubMed  Google Scholar 

  116. Graff, C. P., Chester, K., Begent, R., and Wittrup, K. D. (2004) Directed evolution of an anti-carcinoembryonic antigen scFv with a 4-day monovalent dissociation half-time at 37 degrees C. Protein Eng. Des. Sel. 17, 293–304.

    CAS  PubMed  Google Scholar 

  117. Olafsen, T., Tan, G. J., Cheung, C. W., et al. (2004) Characterization of engineered anti-p185HER-2 (scFv-CH3)2 antibody fragments (minibodies) for tumor targeting. Protein Eng. Des. Sel. 17, 315–323.

    CAS  PubMed  Google Scholar 

  118. Epenetos, A. A., Snook, D., Durbin, H., Johnson, P. M., and Taylor-Papadimitriou, J. (1986) Limitations of radiolabeled monoclonal antibodies for localization of human neoplasms. Cancer Res. 46, 3183–3191.

    CAS  PubMed  Google Scholar 

  119. Neuwelt, E. A., Specht, H. D., Barnett, P. A., et al. (1987) Increased delivery of tumor-specific monoclonal antibodies to brain after osmotic blood-brain barrier modification in patients with melanoma metastatic to the central nervous system. Neurosurgery 20, 885–895.

    CAS  PubMed  Google Scholar 

  120. Jain, R. K., and Baxter, L. T. (1988) Mechanisms of heterogeneous distribution of monoclonal antibodies and other macromolecules in tumors: significance of elevated interstitial pressure. Cancer Res. 48, 7022–7032.

    CAS  PubMed  Google Scholar 

  121. Murayama, O., Nishida, H., and Sekiguchi, K. (1996) Novel peptide ligands for integrin alpha 6 beta 1 selected from a phage display library. J. Biochem. (Tokyo) 120, 445–451.

    CAS  Google Scholar 

  122. Koivunen, E., Wang, B., and Ruoslahti, E. (1995) Phage libraries displaying cyclic peptides with different ring sizes: ligand specificities of the RGD-directed integrins. Biotechnology (NY) 13, 265–270.

    CAS  Google Scholar 

  123. Koivunen, E., Restel, B. H., Rajotte, D., Lahdenranta, J., Hagedorn, M., Arap, W., and Pasqualini, R. (1999) Integrin-binding peptides derived from phage display libraries. Methods Mol. Biol. 129, 3–17.

    CAS  PubMed  Google Scholar 

  124. Koivunen, E., Arap, W., Valtanen, H., et al. (1999) Tumor targeting with a selective gelatinase inhibitor. Nat. Biotechnol. 17, 768–774.

    CAS  PubMed  Google Scholar 

  125. Yanofsky, S. D., Baldwin, D. N., Butler, J. H., et al. (1996) High affinity type I interleukin 1 receptor antagonists discovered by screening recombinant peptide libraries. Proc. Natl. Acad. Sci. USA 93, 7381–7386.

    CAS  PubMed  Google Scholar 

  126. Pennington, M. E., Lam, K. S., and Cress, A. E. (1996) The use of a combinatorial library method to isolate human tumor cell adhesion peptides. Mol. Divers, 2, 19–28.

    CAS  PubMed  Google Scholar 

  127. Goodson, R. J., Doyle, M. V., Kaufman, S. E., and Rosenberg, S. (1994) High-affinity urokinase receptor antagonists identified with bacteriophage peptide display. Proc. Natl. Acad. Sci. USA 91, 7129–7133.

    CAS  PubMed  Google Scholar 

  128. Burg, M. A., Pasqualini, R., Arap, W., Ruoslahti, E., and Stallcup, W. B. (1999) NG2 proteoglycan-binding peptides target tumor neovasculature. Cancer Res. 59, 2869–2874.

    CAS  PubMed  Google Scholar 

  129. Sims, D., Horne, M. M., Creighan, M., and Donald, A. (1994) Heterogeneity of pericyte populations in equine skeletal muscle and dermal microvessels: a quantitative study. Anat. Histol. Embryol. 23, 232–238.

    CAS  PubMed  Google Scholar 

  130. Lindahl, P., and Betsholtz, C. (1998) Not all myofibroblasts are alike: revisiting the role of PDGF-A and PDGF-B using PDGF-targeted mice. Curr. Opin. Nephrol. Hypertens. 7, 21–26.

    CAS  PubMed  Google Scholar 

  131. Hirschi, K. K., and D’Amore, P. A. (1997) Control of angiogenesis by the pericyte: molecular mechanisms and significance. Exs. 79, 419–428.

    Google Scholar 

  132. Schrappe, M., Klier, F. G., Spiro, R. C., Waltz, T. A., Reisfeld, R. A., and Gladson, C. L. (1991) Correlation of chondroitin sulfate proteoglycan expression on proliferating brain capillary endothelial cells with the malignant phenotype of astroglial cells. Cancer Res. 51, 4986–4993.

    CAS  PubMed  Google Scholar 

  133. Leger, O., Johnson-Leger, C., Jackson, E., Coles, B., and Dean, C. (1994) The chondroitin sulfate proteoglycan NG2 is a tumour-specific antigen on the chemically induced rat chondrosarcoma HSN. Int J. Cancer 58, 700–705.

    CAS  PubMed  Google Scholar 

  134. Behm, F. G., Smith, F. O., Raimondi, S. C., Pui, C. H., and Bernstein, I. D. (1996) Human homologue of the rat chondroitin sulfate proteoglycan, NG2, detected by monoclonal antibody 7.1, identifies childhood acute lymphoblastic leukemias with t(4;11)(q21;q23) or t(11;19)(q23;p13) and MLL gene rearrangements. Blood 87, 1134–1139.

    CAS  PubMed  Google Scholar 

  135. Scott, J. K., and Smith, G. P. (1990) Searching for peptide ligands with an epitope library. Science 249, 386–390.

    CAS  PubMed  Google Scholar 

  136. Collins, J., Horn, N., Wadenback, J., and Szardenings, M. (2001) Cosmix-plexing: a novel recombinatorial approach for evolutionary selection from combinatorial libraries. J. Biotechnol. 74, 317–338.

    CAS  PubMed  Google Scholar 

  137. Pasqualini, R., Koivunen, E., and Ruoslahti, E. (1997) Alpha v integrins as receptors for tumor targeting by circulating ligands. Nat. Biotechnol. 15, 542–546.

    CAS  PubMed  Google Scholar 

  138. Pinilla, C., Appel, J. R., Borras, E., and Houghten, R. A. (2003) Advances in the use of synthetic combinatorial chemistry: mixture-based libraries. Nat. Med. 9, 118–122.

    CAS  PubMed  Google Scholar 

  139. Wrighton, N. C., Balasubramanian, P., Barbone, F. P., et al. (1997) Increased potency of an erythropoietin peptide mimetic through covalent dimerization. Nat. Biotechnol. 15, 1261–1265.

    CAS  PubMed  Google Scholar 

  140. Hannon, G. J. (2002) RNA interference. Nature 418, 244–251.

    CAS  PubMed  Google Scholar 

  141. Wang, L., Prakash, R. K., Stein, C. A., Koehn, R. K., and Ruffner, D. E. (2003) Progress in the delivery of therapeutic oligonucleotides: organ/cellular distribution and targeted delivery of oligonucleotides in vivo. Antisense Nucleic Acid Drug Dev. 13, 169–189.

    CAS  PubMed  Google Scholar 

  142. Czauderna, F., Fechtner, M., Dames, S., et al. (2003) Structural variations and stabilising modifications of synthetic siRNAs in mammalian cells. Nucleic Acids Res. 31, 2705–2716.

    CAS  PubMed  Google Scholar 

  143. Reed, R. K., Lepsoe, S., and Wiig, H. (1989) Interstitial exclusion of albumin in rat dermis and subcutis in over-and dehydration. Am. J. Physiol. 257, H1819–H1827.

    CAS  PubMed  Google Scholar 

  144. Krol, A., Maresca, J., Dewhirst, M. W., and Yuan, F. (1999) Available volume fraction of macromolecules in the extravascular space of a fibrosarcoma: implications for drug delivery. Cancer Res. 59, 4136–4141.

    CAS  PubMed  Google Scholar 

  145. Jain, R. K. (1997) Delivery of molecular and cellular medicine to solid tumors. Adv. Drug Deliv. Rev. 26, 71–90.

    CAS  PubMed  Google Scholar 

  146. Vaupel, P., Kallinowski, F., and Okunieff, P. (1989) Blood flow, oxygen and nutrient supply, and metabolic microenvironment of human tumors: a review. Cancer Res. 49, 6449–6465.

    CAS  PubMed  Google Scholar 

  147. Jain, R. K. (1988) Determinants of tumor blood flow: a review. Cancer Res. 48, 2641–2658.

    CAS  PubMed  Google Scholar 

  148. Hwang, K. M., Fodstad, O., Oldham, R. K., and Morgan, A. C., Jr. (1985) Radiolocalization of xenografted human malignant melanoma by a monoclonal antibody (9.2.27) to a melanoma-associated antigen in nude mice. Cancer Res. 45, 4150–4155.

    CAS  PubMed  Google Scholar 

  149. Yang, H. M., and Reisfeld, R. A. (1988) Doxorubicin conjugated with a monoclonal antibody directed to a human melanoma-associated proteoglycan suppresses the growth of established tumor xenografts in nude mice. Proc. Natl. Acad. Sci. USA, 85, 1189–1193.

    CAS  PubMed  Google Scholar 

  150. Yang, H. M., and Reisfeld, R. A. (1988) Pharmacokinetics and mechanism of action of a doxorubicin-monoclonal antibody 9.2.27 conjugate directed to a human melanoma proteoglycan. J. Natl. Cancer Inst. 80, 1154–1159.

    CAS  PubMed  Google Scholar 

  151. Colapinto, E. V., Zalutsky, M. R., Archer, G. E., et al. (1990) Radioimmunotherapy of intracerebral human glioma xenografts with 131I-labeled F(ab’)2 fragments of monoclonal antibody Mel-14. Cancer Res. 50, 1822–1827.

    CAS  PubMed  Google Scholar 

  152. Colapinto, E. V., Humphrey, P. A., Zalutsky, M. R., et al. (1988) Comparative localization of murine monoclonal antibody Me1-14 F(ab’)2 fragment and whole IgG2a in human glioma xenografts. Cancer Res. 48, 5701–5707.

    CAS  PubMed  Google Scholar 

  153. Bigner, D. D., Brown, M., Coleman, R. E., et al. (1995) Phase I studies of treatment of malignant gliomas and neoplastic meningitis with 131I-radiolabeled monoclonal antibodies anti-tenascin 81C6 and anti-chondroitin proteoglycan sulfate Me1-14 F (ab’)2-a preliminary report. J. Neurooncol. 24, 109–122.

    CAS  PubMed  Google Scholar 

  154. Cokgor, I., Akabani, G., Friedman, H. S., et al. (2001) Long term response in a patient with neoplastic meningitis secondary to melanoma treated with (131)I-radiolabeled antichondroitin proteoglycan sulfate Mel-14 F(ab’)(2): a case study. Cancer 91, 1809–1813.

    CAS  PubMed  Google Scholar 

  155. Larsen, R. H., Akabani, G., Welsh, P., and Zalutsky, M. R. (1998) The cytotoxicity and microdosimetry of astatine-211-labeled chimeric monoclonal antibodies in human glioma and melanoma cells in vitro. Radiat. Res. 149, 155–162.

    CAS  PubMed  Google Scholar 

  156. Makagiansar, I. T., Williams, S., Dahlin-Huppe, K., Fukushi, J., Mustelin, T., and Stallcup, W. B. (2004) Phosphorylation of NG2 proteoglycan by protein kinase C-alpha regulates polarized membrane distribution and cell motility. J. Biol. Chem. 279, 55,262–55,270.

    CAS  PubMed  Google Scholar 

  157. Smith, M. R. (2003) Rituximab (monoclonal anti-CD20 antibody): mechanisms of action and resistance. Oncogene 22, 7359–7368.

    CAS  PubMed  Google Scholar 

  158. Lin, C. C., Shen, Y. C., Chuang, C. K., and Liao, S. K. (2001) Analysis of a murine anti-ganglioside GD2 monoclonal antibody expressing both IgG2a and IgG3 isotypes: monoclonality, apoptosis triggering, and activation of cellular cytotoxicity on human melanoma cells. Adv. Exp. Med. Biol. 491, 419–429.

    CAS  PubMed  Google Scholar 

  159. Baselga, J., and Albanell, J. (2001) Mechanism of action of anti-HER2 monoclonal antibodies. Ann. Oncol. 12Suppl 1, S35–S41.

    PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2007 Humana Press Inc.

About this protocol

Cite this protocol

Chekenya, M., Immervoll, H. (2007). The NG2/HMP Proteoglycan as a Cancer Therapeutic Target. In: Sioud, M. (eds) Target Discovery and Validation Reviews and Protocols. Methods in Molecular Biology™, vol 361. Humana Press. https://doi.org/10.1385/1-59745-208-4:93

Download citation

  • DOI: https://doi.org/10.1385/1-59745-208-4:93

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-58829-890-4

  • Online ISBN: 978-1-59745-208-3

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics