Changes in Vascular Permeability and Expression of Different Angiogenic Factors Following Anti-Angiogenic Treatment in Rat Glioma

Background Anti-angiogenic treatments of malignant tumors targeting vascular endothelial growth factor receptors (VEGFR) tyrosine kinase are being used in different early stages of clinical trials. Very recently, VEGFR tyrosine kinase inhibitor (Vetanalib, PTK787) was used in glioma patient in conjunction with chemotherapy and radiotherapy. However, changes in the tumor size, tumor vascular permeability, vascular density, expression of VEGFR2 and other angiogenic factors in response to PTK787 are not well documented. This study was to determine the changes in tumor size, vascular permeability, fractional plasma volume and expression of VEGFR2 in PTK787 treated U-251 glioma rat model by in vivo magnetic resonance imaging (MRI) and single photon emission computed tomography (SPECT). The findings were validated with histochemical and western blot studies. Methodologies and Principal Findings Seven days after implantation of U251 glioma cells, animals were treated with either PTK787 or vehicle-only for two weeks, and then tumor size, tumor vascular permeability transfer constant (Ktrans), fractional plasma volume (fPV) and expression of VEGFR2 and other relevant angiogenic factors were assessed by in vivo MRI and SPECT (Tc-99-HYNIC-VEGF), and by immunohistochemistry and western blot analysis. Dynamic contrast-enhanced MRI (DCE-MRI) using a high molecular weight contrast agent albumin-(GdDTPA) showed significantly increased Ktrans at the rim of the treated tumors compared to that of the central part of the treated as well as the untreated (vehicle treated) tumors. Size of the tumors was also increased in the treated group. Expression of VEGFR2 detected by Tc-99m-HYNIC-VEGF SPECT also showed significantly increased activity in the treated tumors. In PTK787-treated tumors, histological staining revealed increase in microvessel density in the close proximity to the tumor border. Western blot analysis indicated increased expression of VEGF, SDF-1, HIF-1α, VEGFR2, VEGFR3 and EGFR at the peripheral part of the treated tumors compared to that of central part of the treated tumors. Similar expression patters were not observed in vehicle treated tumors. Conclusion These findings indicate that PTK787 treatment induced over expression of VEGF as well as the Flk-1/VEGFR2 receptor tyrosine kinase, especially at the rim of the tumor, as proven by DCE-MRI, SPECT imaging, immunohistochemistry and western blot.

[1]  H. Dvorak,et al.  Vascular permeability factor/vascular endothelial growth factor and the significance of microvascular hyperpermeability in angiogenesis. , 1999, Current topics in microbiology and immunology.

[2]  Gerald E. York,et al.  Creation of DICOM—Aware Applications Using ImageJ , 2005, Journal of Digital Imaging.

[3]  D. Siemann,et al.  Monitoring the treatment efficacy of the vascular disrupting agent CA4P. , 2007, European journal of cancer.

[4]  A. Bikfalvi,et al.  Tumor angiogenesis , 2020, Advances in cancer research.

[5]  C. Haie-meder,et al.  [Use of the functional imaging modalities in radiation therapy treatment planning in patients with glioblastoma]. , 2005, Bulletin du cancer.

[6]  P. Wen,et al.  Emerging antiangiogenic treatments for gliomas - efficacy and safety issues. , 2008, Current opinion in neurology.

[7]  D. Gianfelice,et al.  MR imaging-guided focused US ablation of breast cancer: histopathologic assessment of effectiveness-- initial experience. , 2003, Radiology.

[8]  A Horsman,et al.  Dynamic MR imaging of invasive breast cancer: correlation with tumour grade and other histological factors. , 1997, The British journal of radiology.

[9]  F. Blankenberg,et al.  SPECT and PET imaging of EGF receptors with site-specifically labeled EGF and dimeric EGF. , 2009, Bioconjugate chemistry.

[10]  Fan Zhang,et al.  Cytokine-mediated deployment of SDF-1 induces revascularization through recruitment of CXCR4+ hemangiocytes , 2006, Nature Medicine.

[11]  P. Wen,et al.  Novel anti-angiogenic therapies for malignant gliomas , 2008, The Lancet Neurology.

[12]  R. Knight,et al.  Detection of migration of locally implanted AC133+ stem cells by cellular magnetic resonance imaging with histological findings , 2008, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[13]  Vimalkumar A. Patel,et al.  In vivo tumor angiogenesis imaging with site-specific labeled 99mTc-HYNIC-VEGF , 2006, European Journal of Nuclear Medicine and Molecular Imaging.

[14]  W E Reddick,et al.  MR imaging of tumor microcirculation: Promise for the new millenium , 1999, Journal of magnetic resonance imaging : JMRI.

[15]  T W Redpath,et al.  Baseline MRI delivery characteristics predict change in invasive ductal breast carcinoma PET metabolism as a result of primary chemotherapy administration. , 2006, Annals of oncology : official journal of the European Society for Medical Oncology.

[16]  P. Wen,et al.  Antiangiogenic therapy in malignant gliomas , 2008, Current opinion in oncology.

[17]  Stephen L. Brown,et al.  Model Selection in Magnetic Resonance Imaging Measurements of Vascular Permeability: Gadomer in a 9L Model of Rat Cerebral Tumor , 2006, Journal of cerebral blood flow and metabolism : official journal of the International Society of Cerebral Blood Flow and Metabolism.

[18]  H. Cheng,et al.  Quantifying angiogenesis in VEGF‐enhanced tissue‐engineered bladder constructs by dynamic contrast‐enhanced MRI using contrast agents of different molecular weights , 2007, Journal of magnetic resonance imaging : JMRI.

[19]  Andrea Sbarbati,et al.  Early Antiangiogenic Activity of SU11248 Evaluated In vivo by Dynamic Contrast-Enhanced Magnetic Resonance Imaging in an Experimental Model of Colon Carcinoma , 2005, Clinical Cancer Research.

[20]  J. Mestan,et al.  PTK787/ZK 222584, a novel and potent inhibitor of vascular endothelial growth factor receptor tyrosine kinases, impairs vascular endothelial growth factor-induced responses and tumor growth after oral administration. , 2000, Cancer research.

[21]  A. Ullrich,et al.  SU5416 is a potent and selective inhibitor of the vascular endothelial growth factor receptor (Flk-1/KDR) that inhibits tyrosine kinase catalysis, tumor vascularization, and growth of multiple tumor types. , 1999, Cancer research.

[22]  M. Mrugala,et al.  Bevacizumab for recurrent malignant gliomas: efficacy, toxicity, and patterns of recurrence. , 2009, Neurology.

[23]  Robert C. Brasch,et al.  MRI monitoring of tumor response following angiogenesis inhibition in an experimental human breast cancer model , 2003, European Journal of Nuclear Medicine and Molecular Imaging.

[24]  Sandra Remer,et al.  The challenges of long-term treatment outcomes in adults with malignant gliomas. , 2004, Clinical journal of oncology nursing.

[25]  N. van Bruggen,et al.  Magnetic resonance imaging detects suppression of tumor vascular permeability after administration of antibody to vascular endothelial growth factor. , 1998, Cancer investigation.

[26]  D M Shames,et al.  Mammary carcinoma model: correlation of macromolecular contrast-enhanced MR imaging characterizations of tumor microvasculature and histologic capillary density. , 1996, Radiology.

[27]  E. Voest,et al.  Target practice: lessons from phase III trials with bevacizumab and vatalanib in the treatment of advanced colorectal cancer. , 2007, The oncologist.

[28]  D M Shames,et al.  Measurement of capillary permeability to macromolecules by dynamic magnetic resonance imaging: A quantitative noninvasive technique , 1993, Magnetic resonance in medicine.

[29]  Marina V Backer,et al.  Molecular imaging of VEGF receptors in angiogenic vasculature with single-chain VEGF-based probes , 2007, Nature Medicine.

[30]  Tracy T Batchelor,et al.  AZD2171, a pan-VEGF receptor tyrosine kinase inhibitor, normalizes tumor vasculature and alleviates edema in glioblastoma patients. , 2007, Cancer cell.

[31]  Meiyappan Solaiyappan,et al.  Reduction of vascular and permeable regions in solid tumors detected by macromolecular contrast magnetic resonance imaging after treatment with antiangiogenic agent TNP-470. , 2003, Clinical cancer research : an official journal of the American Association for Cancer Research.

[32]  R. Brasch,et al.  MRI monitoring of Avastin™ antiangiogenesis therapy using B22956/1, a new blood pool contrast agent, in an experimental model of human cancer , 2004, Journal of magnetic resonance imaging : JMRI.

[33]  Martin Bendszus,et al.  PTK787/ZK222584, an inhibitor of vascular endothelial growth factor receptor tyrosine kinases, decreases glioma growth and vascularization. , 2004, Neurosurgery.

[34]  J. Hennig,et al.  PTK787/ZK 222584, a specific vascular endothelial growth factor-receptor tyrosine kinase inhibitor, affects the anatomy of the tumor vascular bed and the functional vascular properties as detected by dynamic enhanced magnetic resonance imaging. , 2002, Cancer research.

[35]  F. Blankenberg,et al.  Direct site-specific labeling of the Cys-tag moiety in scVEGF with technetium 99m. , 2008, Bioconjugate chemistry.

[36]  G Brix,et al.  Pathophysiologic basis of contrast enhancement in breast tumors , 1999, Journal of magnetic resonance imaging : JMRI.

[37]  Andrea Sbarbati,et al.  In vivo mapping of fractional plasma volume (fpv) and endothelial transfer coefficient (Kps) in solid tumors using a macromolecular contrast agent: Correlation with histology and ultrastructure , 2003, International journal of cancer.

[38]  M. Ogan,et al.  Albumin labeled with Gd-DTPA: an intravascular contrast-enhancing agent for magnetic resonance blood pool imaging: preparation and characterization. , 1987, Investigative radiology.

[39]  A. Arbab,et al.  The role of vascular cell adhesion molecule 1/ very late activation antigen 4 in endothelial progenitor cell recruitment to rheumatoid arthritis synovium. , 2007, Arthritis and rheumatism.

[40]  W. Risau,et al.  Mechanisms of angiogenesis , 1997, Nature.

[41]  M. Westphal,et al.  Inhibition of Glioblastoma Angiogenesis and Invasion by Combined Treatments Directed Against Vascular Endothelial Growth Factor Receptor-2, Epidermal Growth Factor Receptor, and Vascular Endothelial-Cadherin , 2005, Clinical Cancer Research.

[42]  O. Nalcioglu,et al.  Effect of vasodilator hydralazine on tumor microvascular random flow and blood volume as measured by intravoxel incoherent motion (IVIM) weighted MRI in conjunction with Gd-DTPA-Albumin enhanced MRI. , 2001, Magnetic resonance imaging.

[43]  M. Wendland,et al.  Vascular permeability during antiangiogenesis treatment: MR imaging assay results as biomarker for subsequent tumor growth in rats. , 2008, Radiology.

[44]  H. Dvorak,et al.  Vascular permeability factor/vascular endothelial growth factor, microvascular hyperpermeability, and angiogenesis. , 1995, The American journal of pathology.

[45]  A. Fischman,et al.  Technetium-99m-human polyclonal IgG radiolabeled via the hydrazino nicotinamide derivative for imaging focal sites of infection in rats. , 1990, Journal of nuclear medicine : official publication, Society of Nuclear Medicine.

[46]  D M Shames,et al.  Correlation of dynamic contrast-enhanced MR imaging with histologic tumor grade: comparison of macromolecular and small-molecular contrast media. , 1998, AJR. American journal of roentgenology.

[47]  Lian Li,et al.  Patlak plots of Gd‐DTPA MRI data yield blood–brain transfer constants concordant with those of 14C‐sucrose in areas of blood–brain opening , 2003, Magnetic resonance in medicine.

[48]  J. MacFall,et al.  Comparison of three physiologically‐based pharmacokinetic models for the prediction of contrast agent distribution measured by dynamic MR imaging , 2008, Journal of magnetic resonance imaging : JMRI.

[49]  U. Sunar,et al.  The therapeutic mechanisms of ranpirnase-induced enhancement of radiation response on A549 human lung cancer. , 2007, In vivo.

[50]  M. Dewhirst,et al.  Inhibition of tumor growth by targeting tumor endothelium using a soluble vascular endothelial growth factor receptor. , 1998, Cell growth & differentiation : the molecular biology journal of the American Association for Cancer Research.