A VEGF-dependent autocrine loop mediates proliferation and capillarogenesis in bone marrow endothelial cells of patients with multiple myeloma

Roberto Ria; Angelo Vacca; Francesca Russo; Teresa Cirulli; Massimo Massaia; Patrizia Tosi; Michele Cavo; Diego Guidolin; Domenico Ribatti; Franco Dammacco

doi:10.1160/TH04-06-0334

Thrombosis and Haemostasis, Table of Contents

Thromb Haemost 2004; 92(06): 1438-1445
DOI: 10.1160/TH04-06-0334

Endothelium and Vascular Development

Schattauer GmbH

A VEGF-dependent autocrine loop mediates proliferation and capillarogenesis in bone marrow endothelial cells of patients with multiple myeloma

Authors

Roberto Ria

¹Department of Internal Medicine and Clinical Oncology
Angelo Vacca

¹Department of Internal Medicine and Clinical Oncology
Francesca Russo

¹Department of Internal Medicine and Clinical Oncology
Teresa Cirulli

¹Department of Internal Medicine and Clinical Oncology
Massimo Massaia

²Division of Hematology, Department of Medicine and Clinical Oncology, San Giovanni Battista Hospital, Torino
Patrizia Tosi

³Institute of Hematology and Clinical Oncology «Lorenzo & Ariosto Seràgnoli», Policlinico S. Orsola, Bologna
Michele Cavo

³Institute of Hematology and Clinical Oncology «Lorenzo & Ariosto Seràgnoli», Policlinico S. Orsola, Bologna
Diego Guidolin

⁴Department of Human Anatomy and Physiology, University of Padua Medical School, Padua, Italy
Domenico Ribatti

⁵Department of Human Anatomy and Histology, University of Bari Medical School, Bari
Franco Dammacco

¹Department of Internal Medicine and Clinical Oncology

Abstract

Summary

The expression/function of vascular endothelial growth factor (VEGF) and its receptor 2 (VEGFR-2/KDR) in multiple myeloma (MM)-associated angiogenesis is under scrutiny. We show here that bone marrow endothelial cells (EC) from 16 patients with MM (MMEC) highly expressed VEGF-A (the main VEGF isoform) and VEGFR-2 at both mRNA and protein level, whereas EC from 14 patients with monoclonal gammopathy unassociated/unattributable (MG[u]) (MG[u]EC) and 12 human umbilical veins (HUVEC) expressed very low mRNAs and/or proteins. MMEC showed constitutive autophosphorylation in both VEGFR-2 and the associated extracellular signal-regulated kinase-2 (ERK-2), whereas this was marginal in MG[u]EC and HUVEC. MMEC proliferated rapidly and formed a closely-knit capillary meshwork on Matrigel. These cell functions were reduced in the other EC. Autophosphorylation, proliferation and capillarogenesis were prevented by a neutralizing antiVEGF-A antibody, and more efficaciously by an anti-VEGFR-2 antibody. Both antibodies had no effect or were poorly effective on the other EC. These findings as a whole suggest the existence of an autocrine loop of VEGF in MMEC. Since this is very likely a mechanism for the amplification of VEGF activity in neovascularization, it would constitute an appropriate target for antiangiogenic management in MM.

Keywords

Angiogenesis - autocrine growth - endothelial cells - multiple myeloma - VEGF

Full Text

References

References
1 Vacca A, Ribatti D, Roncali L. et al. Bone marrow angiogenesis and progression in multiple myeloma. Br J Haematol 1994; 87: 503-8.
2 Rajkumar SV, Mesa RA, Fonseca R. et al. Bone marrow angiogenesis in 400 patients with monoclonal gammopathy of undetermined significance, multiple myeloma, and primary amyloidosis. Clin Cancer Res 2002; 08: 2210-16.
3 Vacca A, Ria R, Semeraro F. et al. Endothelial cells in the bone marrow of patients with multiple myeloma. Blood 2003; 102: 3340-8.
4 Bellamy WT, Richter L, Frutiger Y. et al. Expression of vascular endothelial growth factor and its receptors in hematopoietic malignancies. Cancer Res 1999; 59: 728-33.
5 Vacca A, Ribatti D, Presta M. et al. Bone marrow neovascularization, plasma cell angiogenic potential, and matrix metalloproteinase-2 secretion parallel progression of human multiple myeloma. Blood 1999; 93: 3064-73.
6 Borset M, Hjorth-Hansen H, Seidel C. et al. Hepatocyte growth factor and its receptor cmet in multiple myeloma. Blood 1996; 88: 3998-4004.
7 Asosingh K, De Raeve H, Menu E. et al. Angiogenic switch during 5T2MM murine myeloma tumorigenesis: role of CD45 heterogeneity. Blood 2004; 103: 3131-7.
8 Dankbar B, Padro T, Leo R. et al. Vascular endothelial growth factor and interleukin-6 in paracrine tumor-stromal cell interactions in multiple myeloma. Blood 2000; 95: 2630-6.
9 Ribatti D, Vacca A, Nico B. et al. The role of mast cells in tumour angiogenesis. Br J Haematol 2001; 115: 514-21.
10 Ria R, Roccaro AM, Merchionne F. et al. Vascular endothelial growth factor and its receptors in multiple myeloma. Leukemia 2003; 17: 1961-6.
11 Gualandris A, Rusnati M, Belleri M. et al. Basic fibroblast growth factor overexpression in endothelial cells: an autocrine mechanism for angiogenesis and angioproliferative diseases. Cell Growth Differ 1996; 07: 147-60.
12 Masood R, Cai J, Zheng T. et al. Vascular endothelial growth factor/vascular permeability factor is an autocrine growth factor for AIDS-Kaposi sarcoma. Proc Natl Acad Sci U S A 1997; 94: 979-84.
13 International Myeloma Working Group. Criteria for the classification of monoclonal gammopathies, multiple myeloma and related disorders: a report of the International Myeloma Working Group. Br J Haematol 2003; 121: 749-57.
14 Durie BGM, Salmon SE. Multiple myeloma, macroglobulinemia and monoclonal gammapathies. In: Hoffbrand AV, Brain MC, Hirsh J. eds. Recent Advances in Hematology. New York, NY: Churchill Livingstone; 1977: 243-61.
15 Vacca A, Ria R, Presta M. et al. alpha(v) beta(3) integrin engagement modulates cell adhesion, proliferation, and protease secretion in human lymphoid tumor cells. Exp Hematol 2001; 29: 993-1003.
16 Guidolin D, Vacca A, Nussdorfer GG. et al. A new image analysis method based on topological and fractal parameters to evaluate the angiostatic activity of docetaxel by using the Matrigel assay in vitro. Microvasc Res 2004; 67: 117-24.
17 Carmeliet P. Mechanisms of angiogenesis and arteriogenesis. Nat Med 2000; 06: 389-95.
18 Streubel B, Chott A, Huber D. et al. Lymphoma-specific genetic aberrations in microvascular endothelial cells in B-cell lymphomas. N Engl J Med 2004; 351: 250-9.
19 Podar K, Tai YT, Lin BK. et al. Vascular endothelial growth factor-induced migration of multiple myeloma cells is associated with beta 1 integrin- and phosphatidylinositol 3-kinasedependent PKC alpha activation. J Biol Chem 2002; 277: 7875-81.
20 Podar K, Tai YT, Davies FE. et al. Vascular endothelial growth factor triggers signaling cascades mediating multiple myeloma cell growth and migration. Blood 2001; 98: 428-35.
21 Vacca A, Ria R, Ribatti D. et al. A paracrine loop in the vascular endothelial growth factor pathway triggers tumor angiogenesis and growth in multiple myeloma. Haematologica 2003; 88: 176-85.
22 Dias S, Hattori K, Zhu Z. et al. Autocrine stimulation of VEGFR-2 activates human leukemic cell growth and migration. J Clin Invest 2000; 106: 511-21.
23 Dias S, Choy M, Alitalo K. et al. Vascular endothelial growth factor (VEGF)-C signaling through FLT-4 (VEGFR-3) mediates leukemic cell proliferation, survival, and resistance to chemotherapy. Blood 2002; 99: 2179-84.
24 Wu W, Shu X, Hovsepyan H. et al. VEGF receptor expression and signaling in human bladder tumors. Oncogene 2003; 22: 3361-70.
25 Podar K, Catley LP, Tai YT. et al. GW654652, the pan-inhibitor of VEGF receptors, blocks growth and migration of multiple myeloma cells in the bone marrow microenvironment. Blood 2004; 103: 3474-9.
26 Lin B, Podar K, Gupta D. et al. The vascular endothelial growth factor receptor tyrosine kinase inhibitor PTK787/ZK222584 inhibits growth and migration of multiple myeloma cells in the bone marrow microenvironment. Cancer Res 2002; 62: 5019-26.
27 Gerber HP, Malik AK, Solar GP. et al. VEGF regulates haematopoietic stem cell survival by an internal autocrine loop mechanism. Nature 2002; 417: 954-8.
28 Waltenberger J, Claesson-Welsh L, Siegbahn A. et al. Different signal transduction properties of KDR and Flt1, two receptors for vascular endothelial growth factor. J Biol Chem 1994; 269: 26988-95.
29 Keyt BA, Nguyen HV, Berleau LT. et al. Identification of vascular endothelial growth factor determinants for binding KDR and FLT-1 receptors. Generation of receptor-selective VEGF variants by site-directed mutagenesis. J Biol Chem 1996; 271: 5638-46.
30 Hideshima T, Richardson P, Anderson KC. Novel therapeutic approaches for multiple myeloma. Immunol Rev 2003; 194: 164-76.