Despite the combined use of surgery, radiotherapy, and chemotherapy to treat glioblastoma multiforme, the survival rate for patients with this aggressive, highly resistant cerebral malignancy remains poor. Recent compelling evidence from cellular and molecular studies of the mechanisms underlying the invasiveness of human gliomas has implicated the matrix metalloproteinase (MMP-9) in the invasion process. We propose here to identify the mechanisms that lead to the overexpression of the gene that encodes this enzyme in human glioblastoma and to inhibit the growth of these tumors by using adenoviral constructs carrying antisense message to MMP-9.
The Specific Aims are as follows:
Specific Aim 1 : Determine the effect of the Ad-MMP-9 antisense construct on glioma cell growth, attachment, migration and invasion in in vitro models. (la): Determine the effect of the Ad-MMP-9 antisense construct on the levels of MMP-9 and other proteases in glioma cells. (lb): Determine and compare the effect of the Ad-MMP-9 antisense construct on glioma cell growth, adhesion, and migration with those of mock and Ad-CMV. (lc): Investigate the effects of the Ad-MMP- 9 antisense construct on the invasive behavior of human glioma cells in in vitro models (Boyden chamber/spheroid assays).
Specific Aim 2 : Determine the in vivo efficacy and toxicity of the Ad-MMP-9 antisense construct. (2a): Determine and compare the effects of the Ad-MMP-9 antisense construct with those of mock or the Ad-CMV construct in inhibiting the invasion and growth of human glioma cell lines injected subcutaneously and intracerebrally in nude mice. (2b): Determine MMP-9 activity during glioma invasion and tumor growth in in vitro and in vivo imaging models using a near-infrared probe specific for MMP-9, and correlate these results with traditional immunohistochemical analysis, in situ hybridization and in vitro enzyme assays. (2c): Evaluate the toxicity of the Ad-MMP-9 antisense construct with that of the Ad-CMV construct given as intracerebral injections in Fischer/Wistar rats.
Specific Aim 3 : Determine the molecular mechanisms that regulate cerebral angiogenesis in relation to the inhibition of MMP-9 in co-cultures of endothelial and glioma cells in both in vitro and in vivo models. (3a): Determine the effect of stable antisense MMP-9 transfectants and infection of Ad-MMP-9 antisense constructs during the formation of capillary-like structures in co-cultures of microvascular endothelial cells with antisense MMP-9 stable cells or glioblastoma cells infected with Ad-MMP-9 antisense construct. (3b): Determine the effects of antisense MMP-9 stable transfectants and Ad-MMP-9 infection on tumor angiogenesis in vivo. We anticipate that these results will substantially augment our understanding of how this overexpressed molecule is inhibited; thus, the information gained should help in developing new therapeutic approaches to treat glioblastomas.

Agency
National Institute of Health (NIH)
Institute
National Institute of Neurological Disorders and Stroke (NINDS)
Type
Research Project (R01)
Project #
5R01NS047699-02
Application #
6790562
Study Section
Cancer Molecular Pathobiology Study Section (CAMP)
Program Officer
Tagle, Danilo A
Project Start
2003-08-15
Project End
2006-04-30
Budget Start
2004-05-01
Budget End
2005-04-30
Support Year
2
Fiscal Year
2004
Total Cost
$259,134
Indirect Cost
Name
University of Illinois at Chicago
Department
Other Basic Sciences
Type
Schools of Medicine
DUNS #
098987217
City
Chicago
State
IL
Country
United States
Zip Code
60612
Zhuang, Thompson; Chelluboina, Bharath; Ponnala, Shivani et al. (2013) Involvement of nitric oxide synthase in matrix metalloproteinase-9- and/or urokinase plasminogen activator receptor-mediated glioma cell migration. BMC Cancer 13:590
Veeravalli, Krishna Kumar; Rao, Jasti S (2012) MMP-9 and uPAR regulated glioma cell migration. Cell Adh Migr 6:509-12
Veeravalli, Krishna Kumar; Ponnala, Shivani; Chetty, Chandramu et al. (2012) Integrin ?9?1-mediated cell migration in glioblastoma via SSAT and Kir4.2 potassium channel pathway. Cell Signal 24:272-81
Ponnala, Shivani; Chetty, Chandramu; Veeravalli, Krishna Kumar et al. (2012) Metabolic remodeling precedes mitochondrial outer membrane permeabilization in human glioma xenograft cells. Int J Oncol 40:509-18
Asuthkar, Swapna; Velpula, Kiran Kumar; Chetty, Chandramu et al. (2012) Epigenetic regulation of miRNA-211 by MMP-9 governs glioma cell apoptosis, chemosensitivity and radiosensitivity. Oncotarget 3:1439-54
Chetty, Chandramu; Vanamala, Sravan K; Gondi, Christopher S et al. (2012) MMP-9 induces CD44 cleavage and CD44 mediated cell migration in glioblastoma xenograft cells. Cell Signal 24:549-59
Ponnala, Shivani; Chetty, Chandramu; Veeravalli, Krishna Kumar et al. (2011) MMP-9 silencing regulates hTERT expression via ?1 integrin-mediated FAK signaling and induces senescence in glioma xenograft cells. Cell Signal 23:2065-75
Ponnala, Shivani; Veeravalli, Krishna Kumar; Chetty, Chandramu et al. (2011) Regulation of DNA repair mechanism in human glioma xenograft cells both in vitro and in vivo in nude mice. PLoS One 6:e26191
Veeravalli, Krishna Kumar; Chetty, Chandramu; Ponnala, Shivani et al. (2010) MMP-9, uPAR and cathepsin B silencing downregulate integrins in human glioma xenograft cells in vitro and in vivo in nude mice. PLoS One 5:e11583
Chetty, Chandramu; Lakka, Sajani S; Bhoopathi, Praveen et al. (2010) Urokinase plasminogen activator receptor and/or matrix metalloproteinase-9 inhibition induces apoptosis signaling through lipid rafts in glioblastoma xenograft cells. Mol Cancer Ther 9:2605-17

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