The central nervous system (CNS) microvasculature differs morphologically and functionally from that of other organs. The CNS is the dose-limiting tissue in the radiotherapeutic management of a wide variety of tumors. Alleviating or protecting against radiation-induced CNS injury would thus be of obvious advantage in cancer treatment. Among the most significant clinical manifestations or radiation toxicity in the CNS are severe alterations in the structural and functional integrity of the brain microvasculature, which may lead to abnormal glial- and endothelial-cell proliferation, fibrosis, edema, and possibly necrosis. The proposed research is based on the hypothesis that (a) the recovery/repair response of the CNS to radiation dependence on gene induction or reduction of specific proteases and inhibitors, and (b) protease/inhibitor activation regulates the astrocyte/glial-endothelial cell interaction.
Our Specific Aims are: (1) Define the mechanisms responsible for radiation-induced uPA, uPAR, and PN-I by using in vitro solo and co-cultures of human astrocytes, glial, and cerebral microvascular endothelial cells. These investigations will include determinations of transcriptional activity, mRNA stability, and active protein levels. First they will determine in vitro, the gene and protein expression of uPA, uPAR and PN-I, which can be induced by radiation in solo and co-cultures of these cells by ELISA, western blotting and northern blotting. Second, they will determine the effect of various pharmacological inhibitors that downregulate these molecules. Finally in addition to pharmacologic inhibitors that affect these molecules, they will also target these molecules by antisense approach (obtain stable transfectants for uPA/uPAR in these cell lines) or antisense sequences delivered via the recombinant adenoviral system which is already established in their laboratory. The effects of these direct and indirect reagents that regulate these molecules will also be assessed in combination with radiation to determine if they exert a synergistic or additive effect on capillary-like structure formation. (2) Determine the effects of induced or decreased gene expression of uPA, uPAR, and PN-I on the radioresponse in in vivo models, and assess the effect of the antisense stable transfectants and inhibitor specified in Specific Aim 1 on tumor angiogenesis in vivo. These investigations will first focus on determining whether the effects in the brain microvasculature correlate with the results of in vitro experiments. Next they will determine the effect of antisense stable transfectants, AEBSF and recombinant adenoviral antisense constructs for uPA/uPAR with and without irradiation, on tumor angiogenesis in vivo. The proposed studies should generate insight not only into the pathogenesis of radiation-induced CNS injury, but also into the processes necessary for successful recovery. This information in turn should suggest novel targets for therapeutic intervention.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
7R01CA085216-02
Application #
6433698
Study Section
Pathology B Study Section (PTHB)
Project Start
2000-05-01
Project End
2005-01-31
Budget Start
2001-04-10
Budget End
2002-01-31
Support Year
2
Fiscal Year
2000
Total Cost
$245,495
Indirect Cost
Name
University of Illinois at Chicago
Department
Other Basic Sciences
Type
Schools of Medicine
DUNS #
121911077
City
Chicago
State
IL
Country
United States
Zip Code
60612
Lakka, Sajani S; Gondi, Christopher S; Rao, Jasti S (2005) Proteases and glioma angiogenesis. Brain Pathol 15:327-41
Pulukuri, Sai MuraliKrishna; Gondi, Christopher S; Lakka, Sajani S et al. (2005) RNA interference-directed knockdown of urokinase plasminogen activator and urokinase plasminogen activator receptor inhibits prostate cancer cell invasion, survival, and tumorigenicity in vivo. J Biol Chem 280:36529-40
Yanamandra, Niranjan; Kondraganti, Shakuntala; Gondi, Christopher S et al. (2005) Recombinant adeno-associated virus (rAAV) expressing TFPI-2 inhibits invasion, angiogenesis and tumor growth in a human glioblastoma cell line. Int J Cancer 115:998-1005
Pulukuri, Sai Murali Krishna; Rao, Jasti S (2005) Activation of p53/p21Waf1/Cip1 pathway by 5-aza-2'-deoxycytidine inhibits cell proliferation, induces pro-apoptotic genes and mitogen-activated protein kinases in human prostate cancer cells. Int J Oncol 26:863-71
Yanamandra, Niranjan; Gumidyala, Krishna V; Waldron, Kevin G et al. (2004) Blockade of cathepsin B expression in human glioblastoma cells is associated with suppression of angiogenesis. Oncogene 23:2224-30
Gondi, Christopher S; Lakka, Sajani S; Yanamandra, Niranjan et al. (2004) Adenovirus-mediated expression of antisense urokinase plasminogen activator receptor and antisense cathepsin B inhibits tumor growth, invasion, and angiogenesis in gliomas. Cancer Res 64:4069-77
Gondi, Christopher S; Lakka, Sajani S; Dinh, Dzung H et al. (2004) RNAi-mediated inhibition of cathepsin B and uPAR leads to decreased cell invasion, angiogenesis and tumor growth in gliomas. Oncogene 23:8486-96
Lakka, Sajani S; Gondi, Christopher S; Yanamandra, Niranjan et al. (2004) Inhibition of cathepsin B and MMP-9 gene expression in glioblastoma cell line via RNA interference reduces tumor cell invasion, tumor growth and angiogenesis. Oncogene 23:4681-9
Yanamandra, Niranjan; Kondraganti, Shakuntala; Srinivasula, Srinivasa M et al. (2004) Activation of caspase-9 with irradiation inhibits invasion and angiogenesis in SNB19 human glioma cells. Oncogene 23:2339-46
Chandrasekar, Nirmala; Mohanam, Sanjeeva; Gujrati, Meena et al. (2003) Downregulation of uPA inhibits migration and PI3k/Akt signaling in glioblastoma cells. Oncogene 22:392-400

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