Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Article
  • Published:

Anti-cancer activity of targeted pro-apoptotic peptides

Nature Medicine volume 5, pages 1032–1038 (1999)Cite this article

Abstract

We have designed short peptides composed of two functional domains, one a tumor blood vessel 'homing' motif and the other a programmed cell death-inducing sequence, and synthesized them by simple peptide chemistry. The 'homing' domain was designed to guide the peptide to targeted cells and allow its internalization. The pro-apoptotic domain was designed to be nontoxic outside cells, but toxic when internalized into targeted cells by the disruption of mitochondrial membranes. Although our prototypes contain only 21 and 26 residues, they were selectively toxic to angiogenic endothelial cells and showed anti-cancer activity in mice. This approach may yield new therapeutic agents.

Access through your institution
Buy or subscribe

This is a preview of subscription content, access via your institution

Access options

Access through your institution

Buy this article

  • Purchase on SpringerLink
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1: Computer-generated model and amino-acid sequence of CNGRC-GGD(KLAKLAK)2.
Figure 2: D(KLAKLAK)2 disrupts mitochondrial membranes.
Figure 3: CNGRC-GG-D(KLAKLAK)2 and (RGD-4C)-GG-D(KLAKLAK)2 induce apoptosis.
Figure 4: CNGRC-GG-D(KLAKLAK)2 induces apoptosis and mitochondrial swelling in DMECs.
Figure 5: Electron microscopic studies of cultured cells.
Figure 6: Treatment of nude mice bearing MDA-MB-435-derived human breast carcinoma xenografts with CNGRC-GG-D(KLAKLAK)2.
Figure 7: Treatment of nude mice bearing MDA-MB-435-derived human breast carcinoma xenografts with (RGD-4C)-GG-D(KLAKLAK)2.

Similar content being viewed by others

References

  1. Risau, W. Mechanisms of angiogenesis. Nature 386, 671–674 (1997).

    Article  CAS  Google Scholar 

  2. Zetter, B.R. Angiogenesis and tumor metastasis. Annu. Rev. Med. 49, 407–424 (1998).

    Article  CAS  Google Scholar 

  3. Bicknell, R. in Tumour Angiogenesis. (eds. Bicknell, R., Lewis, C.E. & Ferrara, N.) 19–28 (Oxford University Press, Oxford 1997).

    Google Scholar 

  4. Folkman, J. in Cancer: Principles and Practice of Oncology. (eds. DeVita, V.T., Hellman, S. & Rosenberg, S.A.) 3075–3087 (Lippincott-Raven, New York, 1997).

    Google Scholar 

  5. Pasqualini, R., Koivunen, E. & Ruoslahti, E. αv integrins as receptors for tumor targeting by circulating ligands. Nature Biotechnol. 15, 542–546 (1997).

    Article  CAS  Google Scholar 

  6. Arap, W., Pasqualini, R. & Ruoslahti, E. Chemotherapy targeted to tumor vasculature. Curr. Opin. Oncol. 10, 560–565 (1998).

    Article  CAS  Google Scholar 

  7. Pasqualini, R., Arap, W., Rajotte, D. & Ruoslahti, E. in I>Phage Display of Proteins and Peptides (eds. Barbas, C., Burton, D., Silverman, G. & Scott, J.) (Cold Spring Harbor, New York, in the press).

  8. Arap, W., Pasqualini, R. & Ruoslahti, E. Cancer treatment by targeted drug delivery to tumor vasculature in a mouse model. Science 279, 377–380 (1998).

    Article  CAS  Google Scholar 

  9. Bredesen D.E. et al. P75(NTR) and the concept of cellular dependence – seeing how the other half die. Cell Death Differ. 5 , 365–371 (1998).

    Article  CAS  Google Scholar 

  10. Ellerby, H.M. et al. Establishment of a cell-free system of neuronal apoptosis: comparison of premitochondrial, mitochondrial, and postmitochondrial phases. J. Neurosci. 17, 6165– 6178 (1997).

    Article  CAS  Google Scholar 

  11. Mehlen, P. et al. The DCC gene product induces apoptosis by a mechanism requiring receptor proteolysis. Nature 395, 801– 804 (1998).

    Article  CAS  Google Scholar 

  12. Bessalle, R., Kapitkovsky, A., Gorea, A., Shalit, I. & Fridkin, M. All-D-magainin: chirality, antimicrobial activity and proteolytic resistance. FEBS Lett. 274 , 151–155 (1990).

    Article  CAS  Google Scholar 

  13. Javadpour, M.M. et al. De novo antimicrobial peptides with low mammalian cell toxicity. J. Med. Chem. 39, 3107– 3113 (1996).

    Article  CAS  Google Scholar 

  14. Blondelle, S.E. & Houghten, R.A. Design of model amphipathic peptides having potent antimicrobial activities. Biochemistry 31, 12688–12694 ( 1992).

    Article  CAS  Google Scholar 

  15. Epand, R.M. in The Amphipathic Helix (CRC, Boca Raton, Florida, 1993 ).

    Google Scholar 

  16. de Kroon, A., Dolis, D., Mayer, A., Lill, R. & de Kruijff, B. Phospholipid composition of highly purified mitochondrial outer membranes of rat liver and Neurospora crassa. Is cardiolipin present in the mitochondrial outer membrane? Biochim. Biophys. Acta 1325, 108–116 (1997).

    Article  CAS  Google Scholar 

  17. Matsuzaki, K., Murase, O., Fujii, N. & Miyajima, K. Translocation of a channel-forming antimicrobial peptide, magainin 2, across lipid bilayers by forming a pore. Biochemistry 34, 6521 –6526 (1995).

    Article  CAS  Google Scholar 

  18. Hovius, R., Thijssen, J., van der Linden, P., Nicolay, K. & de Kruijff, B. Phospholipid asymmetry of the outer membrane of rat liver mitochondria: Evidence for the presence of cardiolipin on the outside of the outer membrane. FEBS Lett. 330 , 71–76 (1993).

    Article  CAS  Google Scholar 

  19. Baltcheffsky, H., & Baltcheffsky, M. in Mitochondria and Microsomes (eds. Lee, C.P., Schatz, G., Dallner, G.) 519– 540 (Addison-Wesley, Reading, Massachusetts, 1981).

    Google Scholar 

  20. Daum, G. Lipids of Mitochondria. Biochim. Biophys. Acta 882, 1–42 (1985).

    Article  Google Scholar 

  21. Hart, S.L. et al. Cell binding and internalization by filamentous phage displaying a cyclic Arg-Gly-Asp-containing peptide. J. Biol. Chem. 269, 12468–12474 (1994).

    CAS  PubMed  Google Scholar 

  22. Bretscher, M.S. Endocytosis and recycling of the fibronectin receptor in CHO cells. EMBO J. 8, 1341–1348 ( 1989).

    Article  CAS  Google Scholar 

  23. Dathe, M. et al. Hydrophobicity, hydrophobic moment, and angle subtended by charged residues modulate antibacterial and haemolytic activity of amphipathic helical peptides. FEBS Lett. 403, 208– 212 (1997).

    Article  CAS  Google Scholar 

  24. Alvarez-Bravo, J., Kurata, S. & Natori, S. Novel synthetic antimicrobial peptides effective against methicillin-resistant Staphylococcus aureus. Biochem. J. 302, 535–538 ( 1994).

    Article  CAS  Google Scholar 

  25. Alnemri, E.S. et al. ICE/CED-3 protease nomenclature. Cell 87, 171 (1996).

    Article  CAS  Google Scholar 

  26. Hernier, B.G. et al. Characterization of a human Kaposi's sarcoma cell line that induces angiogenic tumors in animals. AIDS 8, 575–581 (1994).

    Article  Google Scholar 

  27. Samaniego, F. et al. Vascular endothelial growth factor and basic fibroblast growth factor present in Kaposi's sarcoma (KS) are induced by inflammatory cytokines and synergize to promote vascular permeability and KS lesion development. Amer. J. Path. 152, 1433– 1443 (1998).

    CAS  PubMed  Google Scholar 

  28. Goto, F., Goto, K., Weindel, K. & Folkman, J. Synergistic effects of vascular endothelial growth factor and basic fibroblast growth factor on the proliferation and cord formation of bovine capillary endothelial cells within collagen gels. Lab. Invest. 69, 508 –517 (1993).

    CAS  PubMed  Google Scholar 

  29. Wade, D. et al. All-D amino acid-containing channel-forming antibiotic peptides. Proc. Natl Acad. Sci. USA 87, 4761– 4765 (1990).

    Article  CAS  Google Scholar 

  30. Mancheno, J.M., Martinez del Pozo, A., Albar, J.P., Onaderra, M. & Gavilanes, J.G. A peptide of nine amino acid residues from α*-sarcin cytotoxin is a membrane-perturbing structure. J. Peptide Res. 51, 142– 148 (1998).

    Article  CAS  Google Scholar 

  31. Pasqualini, R. & Ruoslahti, E. Organ targeting in vivo using phage display peptide libraries. Nature 380, 36–366 (1996).

    Article  Google Scholar 

Download references

Acknowledgements

We thank W.K. Cavenee and G. Salvesen for comments and critical reading of the manuscript. This work was supported by grants CA74238, CA28896 (to ER) NS33376 and Cancer Center support grant CA30199 (to R.P., D.B. and E.R.) from the National Cancer Institute (USA), and DAMD17-98-1-8581 (to D.B. and R.P.) from the DOD-PCRP. H.M.E. is the recipient of a NS10050 NRSA senior fellowship grant. W.A. is the recipient of a CaP CURE award.

Author information

Author notes

  1. H. Michael Ellerby, Lisa M. Ellerby, Gabriel Del Rio, Rammohan Rao & Dale E. Bredesen

    Present address: The Buck Center for Research in Aging, 8001 Redwood Blvd, Novato, California, 94945, USA

  2. Renate Kain

    Present address: Clinical Institute for Clinical Pathology, Dept. Ultrastructural Pathology and Cell Biology, University of Vienna/AKH Wien, Währinger Gürtel 18-20, A-1090, Wien, Austria

  3. Wadih Arap & Renata Pasqualini

    Present address: The University of Texas M.D. Anderson Cancer Center, 1515 Holcombe Boulevard, Houston, Texas, 77030, USA

Authors and Affiliations

  1. Program on Aging and Cancer and Program on Cell Adhesion, The Burnham Institute, 10901 North Torrey Pines Rd., La Jolla, 92037, California, USA

    H. Michael Ellerby, Wadih Arap, Lisa M. Ellerby, Renate Kain, Rebecca Andrusiak, Gabriel Del Rio, Stanislaw Krajewski, Christian R. Lombardo, Rammohan Rao, Erkki Ruoslahti, Dale E. Bredesen & Renata Pasqualini

Authors
  1. H. Michael Ellerby
    View author publications

    You can also search for this author inPubMed Google Scholar

  2. Wadih Arap
    View author publications

    You can also search for this author inPubMed Google Scholar

  3. Lisa M. Ellerby
    View author publications

    You can also search for this author inPubMed Google Scholar

  4. Renate Kain
    View author publications

    You can also search for this author inPubMed Google Scholar

  5. Rebecca Andrusiak
    View author publications

    You can also search for this author inPubMed Google Scholar

  6. Gabriel Del Rio
    View author publications

    You can also search for this author inPubMed Google Scholar

  7. Stanislaw Krajewski
    View author publications

    You can also search for this author inPubMed Google Scholar

  8. Christian R. Lombardo
    View author publications

    You can also search for this author inPubMed Google Scholar

  9. Rammohan Rao
    View author publications

    You can also search for this author inPubMed Google Scholar

  10. Erkki Ruoslahti
    View author publications

    You can also search for this author inPubMed Google Scholar

  11. Dale E. Bredesen
    View author publications

    You can also search for this author inPubMed Google Scholar

  12. Renata Pasqualini
    View author publications

    You can also search for this author inPubMed Google Scholar

Corresponding authors

Correspondence to Erkki Ruoslahti, Dale E. Bredesen or Renata Pasqualini.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Ellerby, H., Arap, W., Ellerby, L. et al. Anti-cancer activity of targeted pro-apoptotic peptides. Nat Med 5, 1032–1038 (1999). https://doi.org/10.1038/12469

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1038/12469

Search

Advanced search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing