Targeting tumor vasculature with novel Listeria-based vaccines directed against CD105

  • Laurence M. Wood
  • Zhen-Kun Pan
  • Patrick Guirnalda
  • Peter Tsai
  • Matthew Seavey
  • Yvonne Paterson
Original article

Abstract

The FDA approval of bevacizumab (Avastin®, Genentech/Roche), a monoclonal antibody raised against human VEGF-A, as second-line therapy for colon and lung carcinoma validated the approach of targeting human tumors with angiogenesis inhibitors. While the VEGF/VEGFR pathway is a viable target for anti-angiogenesis tumor therapy, additional targets involved in tumor neovascularization have been identified. One promising target present specifically on tumor vasculature is endoglin (CD105), a member of the TGF-β receptor complex expressed on vascular endothelium and believed to play a role in angiogenesis. Monoclonal antibody therapy and preventive vaccination against CD105 has met with some success in controlling tumor growth. This report describes the in vivo proof-of-concept studies for two novel therapeutic vaccines, Lm-LLO-CD105A and Lm-LLO-CD105B, directed against CD105 as a strategy to target neovascularization of established tumors. Listeria-based vaccines directed against CD105 lead to therapeutic responses against primary and metastatic tumors in the 4T1-Luc and NT-2 mouse models of breast cancer. In a mouse model for autochthonous Her-2/neu-driven breast cancer, Lm-LLO-CD105A vaccination prevented tumor incidence in 20% of mice by week 58 after birth while all control mice developed tumors by week 40. In comparison with previous Listeria-based vaccines targeting tumor vasculature, Lm-LLO-CD105A and Lm-LLO-CD105B demonstrated equivalent or superior efficacy against two transplantable mouse models of breast cancer. Support is provided for epitope spreading to endogenous tumor antigens and reduction in tumor vascularity after vaccination with Listeria-based CD105 vaccines. Reported here, these CD105 therapeutic vaccines are highly effective in stimulating anti-angiogenesis and anti-tumor immune responses leading to therapeutic efficacy against primary and metastatic breast cancer.

Keywords

Endoglin CD105 Breast cancer Vasculature Listeria monocytogenes Vaccine 

Notes

Conflict of interest

Yvonne Paterson wishes to disclose that she has a financial interest in Advaxis, a vaccine and therapeutic company that has licensed or has an option to license all patents from the University of Pennsylvania that concern the use of Listeria monocytogenes or Listerial products as vaccines.

Supplementary material

262_2011_1002_MOESM1_ESM.eps (447 kb)
Characterization of CD105-specific responses. To characterize the CD105-specific immune response generated by the attenuated Listeria-based CD105 vaccine, Lm-LLO-CD105, 20-mer peptides were generated from the CD105A fragment and assayed for their ability to stimulate IFN-γ secretion by CD8+ T cell enriched splenocytes. a Depiction of 20-mer peptides from the CD105 sequence present in Lm-LLO-CD105A containing possible MHC Class I epitopes. b IFN-γ ELISpot analysis of CD105-specific CD8+ T cell responses in unvaccinated or Lm-LLO-CD105A-vaccinated mice was performed. Briefly, CD8+ T cell enriched splenocytes were co-cultured overnight with irradiated splenocytes loaded with overlapping peptides depicted in Fig. 4A along with positive control H-2kD LLO peptide and either IL-2 or media alone. IFN-γ-producing splenocytes after overnight stimulation were counted by on an ELISpot reader. Supplementary material 1 (EPS 447 kb)

References

  1. 1.
    Algire GH, Chalkley HW, Legallais FY, Park HD (1945) Vascular reactions of normal and malignant tissues in vivo. I. Vascular reactions of mice to wounds and to normal and neoplastic transplants. J Natl Cancer Inst 11(3):555–580Google Scholar
  2. 2.
    Youngner JS, Algire GH (1949) The effect of vascular occlusion on transplanted tumors. J Natl Cancer Inst 10(3):565–579PubMedGoogle Scholar
  3. 3.
    Folkman J (1971) Tumor angiogenesis: therapeutic implications. N Engl J Med 285(21):1182–1186PubMedCrossRefGoogle Scholar
  4. 4.
    Folkman J, Merler E, Abernathy C, Williams G (1971) Isolation of a tumor factor responsible for angiogenesis. J Exp Med 133(2):275–288PubMedCrossRefGoogle Scholar
  5. 5.
    New treatments for colorectal cancer (2004) FDA Consum 38(3):17Google Scholar
  6. 6.
    Seavey MM, Maciag PC, Al-Rawi N, Sewell D, Paterson Y (2009) An anti-vascular endothelial growth factor receptor 2/fetal liver kinase-1 Listeria monocytogenes anti-angiogenesis cancer vaccine for the treatment of primary and metastatic Her-2/neu + breast tumors in a mouse model. J Immunol 182(9):5537–5546PubMedCrossRefGoogle Scholar
  7. 7.
    Maciag PC, Seavey MM, Pan ZK, Ferrone S, Paterson Y (2008) Cancer immunotherapy targeting the high molecular weight melanoma-associated antigen protein results in a broad antitumor response and reduction of pericytes in the tumor vasculature. Cancer Res 68(19):8066–8075PubMedCrossRefGoogle Scholar
  8. 8.
    Niethammer AG, Xiang R, Becker JC, Wodrich H, Pertl U, Karsten G et al (2002) A DNA vaccine against VEGF receptor 2 prevents effective angiogenesis and inhibits tumor growth. Nat Med 8(12):1369–1375PubMedCrossRefGoogle Scholar
  9. 9.
    Mizutani N, Luo Y, Mizutani M, Reisfeld RA, Xiang R (2004) DNA vaccines suppress angiogenesis and protect against growth of breast cancer metastases. Breast Dis 20:81–91PubMedGoogle Scholar
  10. 10.
    Lee SH, Mizutani N, Mizutani M, Luo Y, Zhou H, Kaplan C et al (2006) Endoglin (CD105) is a target for an oral DNA vaccine against breast cancer. Cancer Immunol Immunother 55(12):1565–1574PubMedCrossRefGoogle Scholar
  11. 11.
    Lastres P, Letamendia A, Zhang H, Rius C, Almendro N, Raab U et al (1996) Endoglin modulates cellular responses to TGF-beta 1. J Cell Biol 133(5):1109–1121PubMedCrossRefGoogle Scholar
  12. 12.
    Li DY, Sorensen LK, Brooke BS, Urness LD, Davis EC, Taylor DG et al (1999) Defective angiogenesis in mice lacking endoglin. Science 284(5419):1534–1537PubMedCrossRefGoogle Scholar
  13. 13.
    Bourdeau A, Faughnan ME, Letarte M (2000) Endoglin-deficient mice, a unique model to study hereditary hemorrhagic telangiectasia. Trends Cardiovasc Med 10(7):279–285PubMedCrossRefGoogle Scholar
  14. 14.
    Li C, Hampson IN, Hampson L, Kumar P, Bernabeu C, Kumar S (2000) CD105 antagonizes the inhibitory signaling of transforming growth factor beta1 on human vascular endothelial cells. FASEB J 14(1):55–64PubMedGoogle Scholar
  15. 15.
    Li C, Guo B, Bernabeu C, Kumar S (2001) Angiogenesis in breast cancer: the role of transforming growth factor beta and CD105. Microsc Res Tech 52(4):437–449PubMedCrossRefGoogle Scholar
  16. 16.
    Cheifetz S, Bellon T, Cales C, Vera S, Bernabeu C, Massague J et al (1992) Endoglin is a component of the transforming growth factor-beta receptor system in human endothelial cells. J Biol Chem 267(27):19027–19030PubMedGoogle Scholar
  17. 17.
    Mokrosinski J, Krajewska WM (2008) TGF beta signalling accessory receptors. Postepy Biochem 54(3):264–273PubMedGoogle Scholar
  18. 18.
    Bernabeu C, Lopez-Novoa JM, Quintanilla M (2009) The emerging role of TGF-beta superfamily coreceptors in cancer. Biochim Biophys Acta 1792(10):954–973PubMedGoogle Scholar
  19. 19.
    Sanchez-Elsner T, Botella LM, Velasco B, Langa C, Bernabeu C (2002) Endoglin expression is regulated by transcriptional cooperation between the hypoxia and transforming growth factor-beta pathways. J Biol Chem 277(46):43799–43808PubMedCrossRefGoogle Scholar
  20. 20.
    Sorensen LK, Brooke BS, Li DY, Urness LD (2003) Loss of distinct arterial and venous boundaries in mice lacking endoglin, a vascular-specific TGFbeta coreceptor. Dev Biol 261(1):235–250PubMedCrossRefGoogle Scholar
  21. 21.
    Arthur HM, Ure J, Smith AJ, Renforth G, Wilson DI, Torsney E et al (2000) Endoglin, an ancillary TGFbeta receptor, is required for extraembryonic angiogenesis and plays a key role in heart development. Dev Biol 217(1):42–53PubMedCrossRefGoogle Scholar
  22. 22.
    Pardali E, van der Schaft DW, Wiercinska E, Gorter A, Hogendoorn PC, Griffioen AW et al (2011) Critical role of endoglin in tumor cell plasticity of Ewing sarcoma and melanoma. Oncogene 30(3):334–345Google Scholar
  23. 23.
    Duwel A, Eleno N, Jerkic M, Arevalo M, Bolanos JP, Bernabeu C et al (2007) Reduced tumor growth and angiogenesis in endoglin-haploinsufficient mice. Tumour Biol 28(1):1–8PubMedGoogle Scholar
  24. 24.
    Perez-Gomez E, Eleno N, Lopez-Novoa JM, Ramirez JR, Velasco B, Letarte M et al (2005) Characterization of murine S-endoglin isoform and its effects on tumor development. Oncogene 24(27):4450–4461PubMedCrossRefGoogle Scholar
  25. 25.
    Perez-Gomez E, Villa-Morales M, Santos J, Fernandez-Piqueras J, Gamallo C, Dotor J et al (2007) A role for endoglin as a suppressor of malignancy during mouse skin carcinogenesis. Cancer Res 67(21):10268–10277PubMedCrossRefGoogle Scholar
  26. 26.
    Chung AS, Lee J, Ferrara N (2010) Targeting the tumour vasculature: insights from physiological angiogenesis. Nat Rev Cancer 10(7):505–514Google Scholar
  27. 27.
    Li C, Guo B, Wilson PB, Stewart A, Byrne G, Bundred N et al (2000) Plasma levels of soluble CD105 correlate with metastasis in patients with breast cancer. Int J Cancer 89(2):122–126PubMedCrossRefGoogle Scholar
  28. 28.
    Takahashi N, Kawanishi-Tabata R, Haba A, Tabata M, Haruta Y, Tsai H et al (2001) Association of serum endoglin with metastasis in patients with colorectal, breast, and other solid tumors, and suppressive effect of chemotherapy on the serum endoglin. Clin Cancer Res 7(3):524–532PubMedGoogle Scholar
  29. 29.
    Wang JM, Kumar S, Pye D, van Agthoven AJ, Krupinski J, Hunter RD (1993) A monoclonal antibody detects heterogeneity in vascular endothelium of tumours and normal tissues. Int J Cancer 54(3):363–370PubMedCrossRefGoogle Scholar
  30. 30.
    Wang JM, Kumar S, Pye D, Haboubi N, al-Nakib L (1994) Breast carcinoma: comparative study of tumor vasculature using two endothelial cell markers. J Natl Cancer Inst 86(5):386–388PubMedCrossRefGoogle Scholar
  31. 31.
    Burrows FJ, Derbyshire EJ, Tazzari PL, Amlot P, Gazdar AF, King SW et al (1995) Up-regulation of endoglin on vascular endothelial cells in human solid tumors: implications for diagnosis and therapy. Clin Cancer Res 1(12):1623–1634PubMedGoogle Scholar
  32. 32.
    Tabata M, Kondo M, Haruta Y, Seon BK (1999) Antiangiogenic radioimmunotherapy of human solid tumors in SCID mice using (125)I-labeled anti-endoglin monoclonal antibodies. Int J Cancer 82(5):737–742PubMedCrossRefGoogle Scholar
  33. 33.
    Benitez J, Ferreras JM, Munoz R, Arias Y, Iglesias R, Cordoba-Diaz M et al (2005) Cytotoxicity of an ebulin l-anti-human CD105 immunotoxin on mouse fibroblasts (L929) and rat myoblasts (L6E9) cells expressing human CD105. Med Chem 1(1):65–70PubMedCrossRefGoogle Scholar
  34. 34.
    Matsuno F, Haruta Y, Kondo M, Tsai H, Barcos M, Seon BK (1999) Induction of lasting complete regression of preformed distinct solid tumors by targeting the tumor vasculature using two new anti-endoglin monoclonal antibodies. Clin Cancer Res 5(2):371–382PubMedGoogle Scholar
  35. 35.
    Guy CT, Webster MA, Schaller M, Parsons TJ, Cardiff RD, Muller WJ (1992) Expression of the neu protooncogene in the mammary epithelium of transgenic mice induces metastatic disease. Proc Natl Acad Sci USA 89(22):10578–10582PubMedCrossRefGoogle Scholar
  36. 36.
    Reilly RT, Gottlieb MB, Ercolini AM, Machiels JP, Kane CE, Okoye FI et al (2000) HER-2/neu is a tumor rejection target in tolerized HER-2/neu transgenic mice. Cancer Res 60(13):3569–3576PubMedGoogle Scholar
  37. 37.
    Aslakson CJ, Miller FR (1992) Selective events in the metastatic process defined by analysis of the sequential dissemination of subpopulations of a mouse mammary tumor. Cancer Res 52(6):1399–1405PubMedGoogle Scholar
  38. 38.
    Tao K, Fang M, Alroy J, Sahagian GG (2008) Imagable 4T1 model for the study of late stage breast cancer. BMC Cancer 8:228PubMedCrossRefGoogle Scholar
  39. 39.
    Singh R, Paterson Y (2007) In the FVB/N HER-2/neu transgenic mouse both peripheral and central tolerance limit the immune response targeting HER-2/neu induced by Listeria monocytogenes-based vaccines. Cancer Immunol Immunother 56(6):927–938PubMedCrossRefGoogle Scholar
  40. 40.
    Muller WJ (1991) Expression of activated oncogenes in the murine mammary gland: transgenic models for human breast cancer. Cancer Metastasis Rev 10(3):217–227PubMedCrossRefGoogle Scholar
  41. 41.
    Paterson Y, Maciag PC (2005) Listeria-based vaccines for cancer treatment. Curr Opin Mol Ther 7(5):454–460PubMedGoogle Scholar
  42. 42.
    Kim SH, Castro F, Paterson Y, Gravekamp C (2009) High efficacy of a Listeria-based vaccine against metastatic breast cancer reveals a dual mode of action. Cancer Res 69(14):5860–5866PubMedCrossRefGoogle Scholar
  43. 43.
    Huang AY, Gulden PH, Woods AS, Thomas MC, Tong CD, Wang W et al (1996) The immunodominant major histocompatibility complex class I- restricted antigen of a murine colon tumor derives from an endogenous retroviral gene product. Proc Natl Acad Sci USA 93(18):9730–9735PubMedCrossRefGoogle Scholar
  44. 44.
    Albelda SM, Muller WA, Buck CA, Newman PJ (1991) Molecular and cellular properties of PECAM-1 (endoCAM/CD31): a novel vascular cell-cell adhesion molecule. J Cell Biol 114(5):1059–1068PubMedCrossRefGoogle Scholar
  45. 45.
    Miller DW, Graulich W, Karges B, Stahl S, Ernst M, Ramaswamy A et al (1999) Elevated expression of endoglin, a component of the TGF-beta-receptor complex, correlates with proliferation of tumor endothelial cells. Int J Cancer 81(4):568–572PubMedCrossRefGoogle Scholar
  46. 46.
    Fonsatti E, Jekunen AP, Kairemo KJ, Coral S, Snellman M, Nicotra MR et al (2000) Endoglin is a suitable target for efficient imaging of solid tumors: in vivo evidence in a canine mammary carcinoma model. Clin Cancer Res 6(5):2037–2043PubMedGoogle Scholar
  47. 47.
    Abe F, Dafferner AJ, Donkor M, Westphal SN, Scholar EM, Solheim JC et al (2010) Myeloid-derived suppressor cells in mammary tumor progression in FVB Neu transgenic mice. Cancer Immunol Immunother 59(1):47–62Google Scholar
  48. 48.
    Donkor MK, Lahue E, Hoke TA, Shafer LR, Coskun U, Solheim JC et al (2009) Mammary tumor heterogeneity in the expansion of myeloid-derived suppressor cells. Int Immunopharmacol 9(7–8):937–948PubMedCrossRefGoogle Scholar
  49. 49.
    Souders NC, Sewell DA, Pan ZK, Hussain SF, Rodriguez A, Wallecha A et al (2007) Listeria-based vaccines can overcome tolerance by expanding low avidity CD8 + T cells capable of eradicating a solid tumor in a transgenic mouse model of cancer. Cancer Immun 7:2PubMedGoogle Scholar
  50. 50.
    Kamba T, Tam BY, Hashizume H, Haskell A, Sennino B, Mancuso MR et al (2006) VEGF-dependent plasticity of fenestrated capillaries in the normal adult microvasculature. Am J Physiol Heart Circ Physiol 290(2):H560–H576PubMedCrossRefGoogle Scholar
  51. 51.
    Kamba T, McDonald DM (2007) Mechanisms of adverse effects of anti-VEGF therapy for cancer. Br J Cancer 96(12):1788–1795PubMedCrossRefGoogle Scholar
  52. 52.
    Cao Y (2009) Tumor angiogenesis and molecular targets for therapy. Front Biosci 14:3962–3973PubMedCrossRefGoogle Scholar
  53. 53.
    Shiozaki K, Harada N, Greco WR, Haba A, Uneda S, Tsai H et al (2006) Antiangiogenic chimeric anti-endoglin (CD105) antibody: pharmacokinetics and immunogenicity in nonhuman primates and effects of doxorubicin. Cancer Immunol Immunother 55(2):140–150PubMedCrossRefGoogle Scholar
  54. 54.
    Peters C, Peng X, Douven D, Pan ZK, Paterson Y (2003) The induction of HIV Gag-specific CD8 + T cells in the spleen and gut-associated lymphoid tissue by parenteral or mucosal immunization with recombinant Listeria monocytogenes HIV Gag. J Immunol 170(10):5176–5187PubMedGoogle Scholar
  55. 55.
    Maciag PC, Radulovic S, Rothman J (2009) The first clinical use of a live-attenuated Listeria monocytogenes vaccine: a Phase I safety study of Lm-LLO-E7 in patients with advanced carcinoma of the cervix. Vaccine 27(30):3975–3983PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2011

Authors and Affiliations

  • Laurence M. Wood
    • 1
  • Zhen-Kun Pan
    • 1
  • Patrick Guirnalda
    • 1
  • Peter Tsai
    • 1
  • Matthew Seavey
    • 1
    • 2
  • Yvonne Paterson
    • 1
  1. 1.Department of MicrobiologyUniversity of PennsylvaniaPhiladelphiaUSA
  2. 2.Cephalon, Inc.West ChesterUSA

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