Defense Date

2010

Document Type

Dissertation

Degree Name

Doctor of Philosophy

Department

Anatomy & Neurobiology

First Advisor

Helen Fillmore

Second Advisor

Linda Phillips

Third Advisor

John Povlishock

Fourth Advisor

Roland Pittman

Fifth Advisor

Zendra Zehner

Abstract

Glioblastoma multiforme (GBM) is an aggressive central nervous system (CNS) cancer characterized by enhanced tumor cell motility, pernicious invasion into the normal brain, extensive tumor-induced angiogenesis, and adaptive resistance to current therapeutic paradigms. One of the difficulties associated with GBM is the ability of the tumor cells to infiltrate normal CNS tissue. Neurosurgeons can remove the primary tumor mass, but peripheral cells that are inaccessible will ultimately result in a secondary lesion that can lead to death. The matrix metalloproteinases (MMP) are well known for their abilities to facilitate processes of cellular motility and invasion through their clearance of extracellular matrix (ECM). A specific member of this family, MMP-1, is not observed in normal brain, yet its expression is a common characteristic of GBM. The various causes of MMP-1 expression, and its consequences in GBM cells are unknown. As such, functional studies were conducted related to the induction of MMP-1 expression via another molecule intrinsic to GBM, nitric oxide (NO). The exposure of GBM cell lines to nanomolar concentrations of NO produced significant inductions of MMP-1 expression and GBM cell motility. The specific removal of MMP-1 with siRNA elicited an abrogation of NO-stimulated motility, suggesting a pathological contribution by this enzyme. Furthermore, recent accumulating evidence suggests that MMP-1 contributes to tumor cell survival and related angiogenesis in other cancer settings. To investigate these capabilities in GBM, cell lines were stably engineered to have either MMP-1 over-expression or knock-down. Both tumor formation and size were significantly reduced with MMP-1 knock-down and significantly increased with over-expression. In a model of GBM cell-induced angiogenesis, the presence of MMP-1 contributed to an angiogenic phenotype. Further angiogenesis studies revealed a significant recruitment of host endothelium to the tumor interstitium in vivo. Proteomic studies suggest that one mechanism by which MMP-1 could influence angiogenesis is through the easement of the anti-angiogenic tissue inhibitor of metalloproteinases-4 (TIMP-4), since the removal of MMP-1 elicits a significant increase in TIMP-4 detection. Altogether, these functional data present MMP-1 as a promising target for future therapeutic investigation, because it is unique to the GBM environment and contributes to key overlapping GBM pathologies.

Rights

© The Author

Is Part Of

VCU University Archives

Is Part Of

VCU Theses and Dissertations

Date of Submission

April 2010

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