Pigment epithelium-derived factor: A potent inhibitor of angiogenesis

Science

Volumn 285, Issue 5425, 1999, Pages 245-248

  Dawson, D W ^a   Volpert, O V ^a   Gillis, P ^a   Crawford, S E ^a   Xu, H J ^b   Benedict, W ^b   Bouck, N P ^a  

^a NORTHWESTERN UNIVERSITY   (United States)

^b UNIVERSITY OF TEXAS   (United States)

Author keywords

[No Author keywords available]

Indexed keywords

ANGIOGENESIS INHIBITOR;

ANGIOGENESIS; ARTICLE; BLINDNESS; CONTROLLED STUDY; HUMAN; HUMAN CELL; PIGMENT EPITHELIUM; PRIORITY JOURNAL; RETINA ISCHEMIA; RETINA NEOVASCULARIZATION; RETINOPATHY;

ANIMALS; ANIMALS, NEWBORN; ANTIBODIES; CATTLE; CELLS, CULTURED; CHEMOTAXIS; CULTURE MEDIA, CONDITIONED; ENDOTHELIAL GROWTH FACTORS; ENDOTHELIUM, VASCULAR; EYE; EYE PROTEINS; HUMANS; LYMPHOKINES; MICE; NEOVASCULARIZATION, PATHOLOGIC; NEOVASCULARIZATION, PHYSIOLOGIC; NERVE GROWTH FACTORS; OXYGEN; PROTEINS; RATS; RETINA; RETINAL NEOVASCULARIZATION; RETINAL VESSELS; RNA, MESSENGER; SERPINS; TUMOR CELLS, CULTURED; VASCULAR ENDOTHELIAL GROWTH FACTOR A; VASCULAR ENDOTHELIAL GROWTH FACTORS;

EID: 0033538517     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.285.5425.245     Document Type: Article

References (45)

1. N. Bouck, V. Stellmach, S. Hsu, Adv. Cancer Res. 69, 135 (1996); D. Hanahan and J. Folkman, Cell 86, 353 (1996).
2. N. Bouck, V. Stellmach, S. Hsu, Adv. Cancer Res. 69, 135 (1996); D. Hanahan and J. Folkman, Cell 86, 353 (1996).
3. J. Folkman, in Molecular Basis of Cancer, J. Mendelsohn, P. M. Howley, M. A. Israel, L. A. Liotta, Eds. (Saunders, Philadelphia, PA, 1995), pp. 206-232; Nature Med. 1, 27 (1995).
4. J. Folkman, in Molecular Basis of Cancer, J. Mendelsohn, P. M. Howley, M. A. Israel, L. A. Liotta, Eds. (Saunders, Philadelphia, PA, 1995), pp. 206-232; Nature Med. 1, 27 (1995).
5. S. Brem et al., Am. J. Ophthalmol. 84, 323 (1977); P. Henkind, Ibid. 85, 287 (1978).
6. S. Brem et al., Am. J. Ophthalmol. 84, 323 (1977); P. Henkind, Ibid. 85, 287 (1978).
7. G. K. Klintworth, Comeal Angiogenesis: A Comprehensive Critical Review (Springer-Verlag, New York, 1991); G. M. Kaminska and J. Y. Niederkorn, Investig. Ophthalmol. Vis. Sci. 34, 222 (1993).
8. G. K. Klintworth, Comeal Angiogenesis: A Comprehensive Critical Review (Springer-Verlag, New York, 1991); G. M. Kaminska and J. Y. Niederkorn, Investig. Ophthalmol. Vis. Sci. 34, 222 (1993).
9. A. P. Schachat and R. B. Murphy, Eds., Retina (Mosby, Baltimore, MD, ed. 2, 1994).
10. H.-J. Xu et al., Cancer Res. 51, 4481 (1991).
11. PEDF was purified from WERI-Rb-27R (6) serum-free conditioned media by sequential steps consisting of dialysis (molecular mass cutoff, 30 kD) against distilled water, 60 to 95% ammonium sulfate precipitation, step elution from lentil lectin Sepharose 4B (Pharmacia) with 0.5 M α-methyl-D-mannopyranoside, and elution from a HiTrap heparin Sepharose column (Pharmacia) with increasing NaCl gradient. Purification was monitored by an endothelial cell migration assay (26), and the yield was 17.5%. Edman degradation of proteolytically derived internal peptides of the protein yielded two unambiguous sequences (TSLEDFYLDEERTVRVPMMXD and IAQL-PLTGXM) (27). A BLAST protein homology search revealed that PEDF contains identical sequences.
12. S. P. Beccerra, in Chemistry and Biology of Serpins, F. C. Church et al., Eds. (Plenum, New York, 1997), pp. 223-237.
13. J. Tombran-Tink, G. J. Chader, L. V. Johnson, Exp. Eye Res. 53, 411 (1991); F. R. Steele et al., Proc. Natl. Acad. Sci. U.S.A. 90, 1526 (1993).
14. J. Tombran-Tink, G. J. Chader, L. V. Johnson, Exp. Eye Res. 53, 411 (1991); F. R. Steele et al., Proc. Natl. Acad. Sci. U.S.A. 90, 1526 (1993).
15. T. Tanawaki, S. P. Becerra, G. J. Chader, J. P. Schwartz, J. Neurochem. 64, 2509 (1995); Y. Sugita et al., J. Neurosci. Res. 49, 710 (1997).
16. T. Tanawaki, S. P. Becerra, G. J. Chader, J. P. Schwartz, J. Neurochem. 64, 2509 (1995); Y. Sugita et al., J. Neurosci. Res. 49, 710 (1997).
17. R. J. Pignolo, V. J. Cristofalo, M. O. Rotenberg, J. Biol. Chem. 268, 8949 (1993); J. Tombran-Tink et al., J. Neurosci. 15, 4992 (1995).
18. R. J. Pignolo, V. J. Cristofalo, M. O. Rotenberg, J. Biol. Chem. 268, 8949 (1993); J. Tombran-Tink et al., J. Neurosci. 15, 4992 (1995).
19. S. P. Becerra et al., J. Biol. Chem. 268, 23148 (1993); S. P. Becerra et al., ibid. 270, 25992 (1995).
20. S. P. Becerra et al., J. Biol. Chem. 268, 23148 (1993); S. P. Becerra et al., ibid. 270, 25992 (1995).
21. D. W. Dawson, O. V. Volpert, P. Gillis, unpublished data.
22. Human PEDF cDNA was engineered by polymerase chain reaction to encode a COOH-terminal hexa-histidine tag, cloned into pCEP4 (Invitrogen), and transfected into human embryonic kidney cells. Recombinant PEDF was purified from the conditioned media with the Xpress Protein Purification System (Invitrogen).
23. See supplemental figures, available at www.sciencemag. org/feature/data/1040070
24. For preparation of stromal extract, corneas were freed of associated epithelium and as much of the endothelium as possible, washed extensively in ice-cold phosphate-buffered saline (PBS, pH 7.4), and minced into small fragments that were incubated for 24 hours in PBS containing 0.5 mM phenylmethane-sulfonyl fluoride. The extract was filter sterilized, stored at -80°C, and tested in migration assays at a final concentration of 10 μg of protein per milliliter.
25. Y.-Q. Wu and S. P. Becerra, Investig. Ophthalmol. Vis. Sci. 37, 1984 (1996).
26. L. P. Aiello et al., N. Engl. J. Med. 331, 1480 (1994); A. P. Adamis et al., Am. J. Ophthalmol. 118, 445 (1994).
27. L. P. Aiello et al., N. Engl. J. Med. 331, 1480 (1994); A. P. Adamis et al., Am. J. Ophthalmol. 118, 445 (1994).
28. N. Ogata et al., Curr. Eye Res. 16, 9 (1997); K. Hayasaka et al., Life Sci. 63, 1089 (1998); S. A. Vinores et al., J. Neuroimmunol. 89, 43 (1998).
29. N. Ogata et al., Curr. Eye Res. 16, 9 (1997); K. Hayasaka et al., Life Sci. 63, 1089 (1998); S. A. Vinores et al., J. Neuroimmunol. 89, 43 (1998).
30. N. Ogata et al., Curr. Eye Res. 16, 9 (1997); K. Hayasaka et al., Life Sci. 63, 1089 (1998); S. A. Vinores et al., J. Neuroimmunol. 89, 43 (1998).
31. L. P. Aiello et al., Proc. Natl. Acad. Sci. U.S.A. 92, 10457 (1995); A. P. Adamis et al., Arch. Ophthalmol. 114, 66 (1996); J. M. Provis et al., Exp. Eye Res. 65, 555 (1997).
32. L. P. Aiello et al., Proc. Natl. Acad. Sci. U.S.A. 92, 10457 (1995); A. P. Adamis et al., Arch. Ophthalmol. 114, 66 (1996); J. M. Provis et al., Exp. Eye Res. 65, 555 (1997).
33. L. P. Aiello et al., Proc. Natl. Acad. Sci. U.S.A. 92, 10457 (1995); A. P. Adamis et al., Arch. Ophthalmol. 114, 66 (1996); J. M. Provis et al., Exp. Eye Res. 65, 555 (1997).
34. L. E. H. Smith et al., Invest. Ophthalmol. Vis. Sci. 35, 101 (1994).
35. E. A. Pierce, E. D. Foley, L. E. Smith, Arch. Ophthalmol. 114, 1219 (1996).
36. S. E. Connolly et al., Microvasc. Res. 36, 275 (1988).
37. M. A. Goldberg, S. P. Dunning, H. F. Bunn, Science 242, 1412 (1988).
38. C. J. Gulledge and M. W. Dewhirst, Anticancer Res. 16, 741 (1996).
39. Migration assays were performed in quadruplicate for each sample with bovine adrenal capillary endothelial cells or human dermal microvascular endothelial cells (Clonetics, San Diego, CA) as described (28). To combine multiple experiments, we first subtracted background migration (Bkgd) toward vehicle (0.1% bovine serum albumin) and then normalized data by setting maximum migration toward inducer alone to 100%. All experiments were repeated two to five times. Statistics were performed on raw data before normalization with the Student's t test. Standard errors were converted to percentages.
40. Single-letter abbreviations for the amino acid residues are as follows: A, Ala; D, Asp; E, Glu; F, Phe; G, Gly; I, Ile; L, Leu; M, Met; P, Pro; Q, Gln; R, Arg; S, Ser; T. Thr; V, Val; X, any amino acid; and Y, Tyr.
41. P. J. Polverini, N. P. Bouck, F. Rastinejad, Methods Enzymol. 198, 440 (1991).
42. PEDF antipeptide antibody (anti-PEDF) was raised in rabbits against a peptide containing PEDF amino acids 327 to 343, conjugated to Keyhole-limpet hemocyanin, and affinity-purified on a peptide column. Polyclonal antisera against bacterial recombinant PEDF/EPC-1 (anti-EPC-1) [B. R. DiPaolo, R. J. Pignolo, V. J. Cristofalo, Exp. Cell Res. 220, 178 (1995)] and the antiangiogenic protein angiostatin [M. S. O'Reilly et al., Cell 79, 315 (1994)] were gifts. Purchased reagents included neutralizing anti-VEGF (Genzyme, Cambridge, MA), pan antibodies to TGFβ, and all angiogenic inducers (R & D Systems, Minneapolis, MN) except lysophosphatidic acid (Sigma). All proteins and antibodies were extensively dialyzed against PBS before use in biological assays.
43. PEDF antipeptide antibody (anti-PEDF) was raised in rabbits against a peptide containing PEDF amino acids 327 to 343, conjugated to Keyhole-limpet hemocyanin, and affinity-purified on a peptide column. Polyclonal antisera against bacterial recombinant PEDF/EPC-1 (anti-EPC-1) [B. R. DiPaolo, R. J. Pignolo, V. J. Cristofalo, Exp. Cell Res. 220, 178 (1995)] and the antiangiogenic protein angiostatin [M. S. O'Reilly et al., Cell 79, 315 (1994)] were gifts. Purchased reagents included neutralizing anti-VEGF (Genzyme, Cambridge, MA), pan antibodies to TGFβ, and all angiogenic inducers (R & D Systems, Minneapolis, MN) except lysophosphatidic acid (Sigma). All proteins and antibodies were extensively dialyzed against PBS before use in biological assays.
44. Human vitreous fluid was withdrawn from three cadaveric eyes (refrigerated within 1.4 to 4.5 hours of death) obtained from individuals without ocular disease. Fluid was frozen until used. Fresh vitreous fluid was obtained from bovine and mouse eyes.
45. We thank A. Mountz for VEGF measurements; B. Kennedy and the Midwest Eye Banks and Transplantation Center for human eye tissue; M. K. Francis and V. Cristofalo for anti-EPC-1; M. O'Reilly and J. Folkman for bovine capillary endothelial cells and angiostatin; and C Hawkins, R. O'Grady, and Y. Mu for assistance with retinoblastomas. Supported by the National Eye Institute, the Retina Research Foundation, the National Cancer Institute, and the Chicago Baseball Charities.