Molecular Regulation of Vascular Abnormalization and Its Role in Glioma

Abstract: Glioblastoma, grade IV glioma, is the one of the deadliest cancers, with a median survival of only 12-15 months despite aggressive treatment including surgery, chemotherapy and radiation. One hallmark of glioblastoma is the morphological and functional abnormalization of tumor blood vessels. The molecular mechanisms involved in this process and their functional and pathological implications are not yet fully understood. Indentification of molecular mechanisms that underlie vascular abnormalization in GBM is necessary to develop efficient treatment regimens for normalizing vascular function.By analyzing the RNA-content of laser microdissected vessels from human biobank specimens using affymetrix microarray analysis, we found that the abnormal glioblastoma vessels have a distinct gene expression signature. We found 95 genes which were differentially expressed in grade IV glioma vessels as compared to vessels in low grade tumors and control brain. 78 of which were up-regulated while 17 were down-regulated. Many of these genes are regulated by VEGFA or TGF? signaling. In addition, we show a significant increase in Smad signaling complexes in the vasculature of human glioblastoma in situ, suggesting that TGF? signaling may play important role in vessel abnormalization.CD93 is a single-pass transmembrane glycoprotein, which we found to be up-regulated in high grade glioma. Vascular expression of CD93 correlates to tumor grade in human glioma. Moreover, high grade glioma patients with high CD93 expression in the vasculature are associate with poor prognosis. We found that knocking down CD93 in endothelial cells with siRNA clearly impaired endothelial cell adhesion, migration and tube formation due to defects in cytoskeletal rearrangement. In addition, tumor growth was severely delayed in the CD93-/- mice.Pleiotrophin, a multi-functional heparin-binding growth factor, promotes glioma growth in several ways. Here, we identify pleiotrophin as a driver of vascular abnormalization in glioma. We found that high pleiotrophin expression correlates with poor survival of patients with astrocytomas. Pleiotrophin overexpression in orthotopic GL261 gliomas increases microvessel density, enhances tumour growth and decreases survival. Vessels in pleiotrophin-expressing gliomas are poorly perfused and display a high degree of abnormality, coinciding with elevated levels of vascular endothelial growth factor (VEGF) deposited in direct proximity to the vasculature. In addition to its role in vessel abnormalization, pleiotrophin enhanced PDGF-B-induced gliomagenesis. Taken together, our results indicate that PTN has an important role in glioma initiation and establishment of the characteristic abnormal tumor vasculature in glioblastoma, identifying PTN as a potential target for therapy.