A BMP7 Variant Inhibits Tumor Angiogenesis In Vitro and In Vivo through Direct Modulation of Endothelial Cell Biology.
Bone morphogenetic proteins (BMPs), members of the TGF-β superfamily, have numerous biological activities including control of growth, differentiation, and vascular development. Using an in vitro co-culture endothelial cord formation assay, we investigated the role of a BMP7 variant (BMP7v) in VEGF, bFGF, and tumor-driven angiogenesis. BMP7v treatment led to disruption of neo-endothelial cord formation and regression of existing VEGF and bFGF cords in vitro. Using a series of tumor cell models capable of driving angiogenesis in vitro, BMP7v treatment completely blocked cord formation. Pre-treatment of endothelial cells with BMP7v significantly reduced their cord forming ability, indicating a direct effect on endothelial cell function. BMP7v activated the canonical SMAD signaling pathway in endothelial cells but targeted gene knockdown using shRNA directed against SMAD4 suggests this pathway is not required to mediate the anti-angiogenic effect. In contrast to SMAD activation, BMP7v selectively decreased ERK and AKT activation, significantly decreased endothelial cell migration and down-regulated expression of critical RTKs involved in VEGF and FGF angiogenic signaling, VEGFR2 and FGFR1 respectively. Importantly, in an in vivo angiogenic plug assay that serves as a measurement of angiogenesis, BMP7v significantly decreased hemoglobin content indicating inhibition of neoangiogenesis. In addition, BMP7v significantly decreased angiogenesis in glioblastoma stem-like cell (GSLC) Matrigel plugs and significantly impaired in vivo growth of a GSLC xenograft with a concomitant reduction in microvessel density. These data support BMP7v as a potent anti-angiogenic molecule that is effective in the context of tumor angiogenesis.
Discovery Research, Eli Lilly and Company, Indianapolis, United States of America.
Courtney M
CM
Tate
Discovery Research, Eli Lilly and Company, Indianapolis, United States of America.
Jacquelyn
J
Mc Entire
Department of Neurosurgery, Università Cattolica del Sacro Cuore, Rome, Italy.
Roberto
R
Pallini
Discovery Research, Eli Lilly and Company, Indianapolis, United States of America.
Eliza
E
Vakana
Discovery Research, Eli Lilly and Company, Indianapolis, United States of America.
Lisa
L
Wyss
Discovery Research, Eli Lilly and Company, Indianapolis, United States of America.
Wayne
W
Blosser
Department of Hematology, Oncology and Molecular Medicine, Istituto Superiore di Sanità, Rome, Italy.
Lucia
L
Ricci-Vitiani
Department of Neurosurgery, Università Cattolica del Sacro Cuore, Rome, Italy.
Quintino Giorgio
QG
D'Alessandris
Institute of Human Anatomy, Università Cattolica del Sacro Cuore, Rome, Italy.
Liliana
L
Morgante
Institute of Human Anatomy, Università Cattolica del Sacro Cuore, Rome, Italy.
Stefano
S
Giannetti
Institute of Pathology, Università Cattolica del Sacro Cuore, Rome, Italy.
Luigi Maria
LM
Larocca
Surgical and Oncological Sciences, University of Palermo, Palermo, Italy.
Matilde
M
Todaro
Surgical and Oncological Sciences, University of Palermo, Palermo, Italy.
Antonina
A
Benfante
Surgical and Oncological Sciences, University of Palermo, Palermo, Italy.
Maria Luisa
ML
Colorito
Surgical and Oncological Sciences, University of Palermo, Palermo, Italy.
Giorgio
G
Stassi
Regina Elena National Cancer Institute, Rome, Italy.
Ruggero
R
De Maria
Discovery Research, Eli Lilly and Company, Indianapolis, United States of America.
Scott
S
Rowlinson
Discovery Research, Eli Lilly and Company, Indianapolis, United States of America.
Louis
L
Stancato
D000273:Adipose Tissue / Q000166:cytology; D000818:Animals; D055419:Bone Morphogenetic Protein 7 / Q000494:pharmacology / Q000627:therapeutic use*; D016923:Cell Death / Q000187:drug effects; D045744:Cell Line, Tumor; D002465:Cell Movement / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D003094:Collagen / Q000494:pharmacology; D004338:Drug Combinations; D042783:Endothelial Cells / Q000187:drug effects / Q000378:metabolism*; D016222:Fibroblast Growth Factor 2 / Q000378:metabolism; D005909:Glioblastoma / Q000098:blood supply*; D061307:Human Umbilical Vein Endothelial Cells / Q000187:drug effects / Q000378:metabolism; D006801:Humans; D007797:Laminin / Q000494:pharmacology; D008297:Male; D008819:Mice, Nude; D014411:Neoplastic Stem Cells / Q000187:drug effects / Q000378:metabolism; D009389:Neovascularization, Pathologic / Q000188:drug therapy* / Q000473:pathology; D018919:Neovascularization, Physiologic / Q000187:drug effects; D011509:Proteoglycans / Q000494:pharmacology; D051496:Receptor, Fibroblast Growth Factor, Type 1 / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects; D051785:Smad Proteins / Q000378:metabolism; D042461:Vascular Endothelial Growth Factor A / Q000378:metabolism; D040301:Vascular Endothelial Growth Factor Receptor-2 / Q000378:metabolism; D023041:Xenograft Model Antitumor Assays
D055419:Bone Morphogenetic Protein 7; D004338:Drug Combinations; D007797:Laminin; D011509:Proteoglycans; D051785:Smad Proteins; D042461:Vascular Endothelial Growth Factor A; D016222:Fibroblast Growth Factor 2; C059485:matrigel; D003094:Collagen; D051496:Receptor, Fibroblast Growth Factor, Type 1; D040301:Vascular Endothelial Growth Factor Receptor-2