title: Cancer antineovascular therapy with liposome drug delivery systems targeted to BiP/GRP78.
abstract: Angiogenesis is crucial for tumor growth and hematogenous metastasis. Specifically expressed and functional protein molecules in angiogenic endothelial cells, especially on the plasma membrane, may be molecular targets for antiangiogenic drugs and drug delivery systems (DDS) in cancer therapy. To discover such target molecules, we performed subcellular proteome analysis of human umbilical vein endothelial cells (HUVECs) treated with or without vascular endothelial growth factor (VEGF) using 2-dimensional difference in-gel electrophoresis (2D-DIGE) and matrix-assisted laser desorption/ionization tandem time-of-flight mass spectrometry (MALDI-TOF/TOF-MS). Among the identified proteins, BiP/GRP78, a molecular chaperone, was highly expressed in the membrane/organelle fraction of HUVECs after VEGF treatment. The involvement of BiP in VEGF-induced angiogenesis was examined by RNA interference. BiP knockdown significantly suppressed VEGF-induced endothelial cell proliferation and VEGF-induced phosphorylation of extracellular-regulated kinase 1/2, phospholipase C-γ, and VEGF receptor-2 in HUVECs. Cell surface biotinylation analysis revealed that the cell surface expression of BiP was elevated in VEGF-activated HUVECs. Aiming to apply BiP to a target molecule in liposomal DDS, we developed liposomes modified with the WIFPWIQL peptide, which has been shown to bind to BiP, and investigated its potential for cancer therapy. The WIFPWIQL-modified liposomes (WIFPWIQL liposomes) were significantly taken up by VEGF-activated HUVECs as compared to peptide-unmodified liposomes. WIFPWIQL liposomes appeared to accumulate in tumor endothelial cells in vivo. WIFPWIQL liposomes containing doxorubicin significantly suppressed tumor growth and prolonged the survival of colon26 NL-17 carcinoma cell-bearing mice. In summary, BiP may regulate VEGF-induced endothelial cell proliferation through VEGFR-2-mediated signaling and be an effective target molecule for cancer antineovascular therapy.
authors.affiliation: Department of Medical Biochemistry, Graduate School of Pharmaceutical Sciences, University of Shizuoka, Suruga-ku, Shizuoka, Japan.
authors.forename: Yasufumi
authors.initials: Y
authors.lastname: Katanasaka
authors.forename: Takayuki
authors.initials: T
authors.lastname: Ishii
authors.forename: Tomohiro
authors.initials: T
authors.lastname: Asai
authors.forename: Hirotaka
authors.initials: H
authors.lastname: Naitou
authors.forename: Noriyuki
authors.initials: N
authors.lastname: Maeda
authors.forename: Fumiaki
authors.initials: F
authors.lastname: Koizumi
authors.forename: Shinichi
authors.initials: S
authors.lastname: Miyagawa
authors.forename: Norio
authors.initials: N
authors.lastname: Ohashi
authors.forename: Naoto
authors.initials: N
authors.lastname: Oku
mesh_terms: D020533:Angiogenesis Inhibitors / Q000008:administration & dosage*; D000818:Animals; D003110:Colonic Neoplasms / Q000098:blood supply* / Q000188:drug therapy* / Q000378:metabolism; D004317:Doxorubicin / Q000008:administration & dosage; D016503:Drug Delivery Systems / Q000379:methods; D015180:Electrophoresis, Gel, Two-Dimensional; D000091342:Endoplasmic Reticulum Chaperone BiP; D042783:Endothelial Cells / Q000187:drug effects / Q000378:metabolism; D055785:Gene Knockdown Techniques; D006360:Heat-Shock Proteins / Q000172:deficiency / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D008081:Liposomes / Q000008:administration & dosage / Q000493:pharmacokinetics; D020935:MAP Kinase Signaling System; D008297:Male; D051379:Mice; D009389:Neovascularization, Pathologic / Q000188:drug therapy; D009842:Oligopeptides / Q000008:administration & dosage; D011471:Prostatic Neoplasms / Q000098:blood supply* / Q000188:drug therapy* / Q000378:metabolism; D040901:Proteomics; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D042461:Vascular Endothelial Growth Factor A / Q000494:pharmacology
chemical_list: D020533:Angiogenesis Inhibitors; D000091342:Endoplasmic Reticulum Chaperone BiP; C000717247:HSPA5 protein, human; D006360:Heat-Shock Proteins; C000717248:Hspa5 protein, mouse; D008081:Liposomes; D009842:Oligopeptides; D042461:Vascular Endothelial Growth Factor A; D004317:Doxorubicin