AdTIMP-2 Inhibits Tumor Growth, Angiogenesis, and Metastasis, and Prolongs Survival in Mice

Abstract

TIMP-2 is a natural matrix metalloproteinase (MMP) inhibitor that prevents the degradation of extracellular matrix proteins. It abolishes the hydrolytic activity of all activated members of the metalloproteinase family and in particular that of MT1-MMP, MMP-2, and MMP-9, which are selective for type IV collagenolysis. Since MMPs have been implicated in both cancer progression and angiogenesis, we generated a recombinant adenovirus to deliver human TIMP-2 (AdTIMP-2) and evaluated its anticancer efficiency in three murine models. Our results demonstrated that overexpression in vitro of TIMP-2 inhibited the invasion of both tumor and endothelial cells without affecting cell proliferation. Its in vivo efficiency has been evaluated in murine lung cancer LLC, and colon cancer C51 in syngeneic mice as well as in human breast cancer MDA-MB231 in athymic mice. Preinfection of tumor cells by AdTIMP-2 resulted in an inhibition of tumor establishment in more than 50% of mice in LLC and C51 models and in 100% mice in the MDA-MB231 model. A single local injection of AdTIMP-2 into preestablished tumors of these three types significantly reduced tumor growth rates by 60--80% and tumor-associated angiogenesis index by 25--75%. Lung metastasis of LLC tumor was inhibited by >90%. In addition, AdTIMP-2-treated mice showed a significantly prolonged survival in all the cancer models tested. These data demonstrate the potential of adenovirus-mediated TIMP-2 therapy in cancer treatment.

The use of tissue inhibitors of matrix metalloproteinases to increase the efficacy of a tumor necrosis factor/interferonγ antitumor therapy

“…The adenoviral doses reported in the literature on in vivo TIMP gene therapy are higher 25,26,28 than the dose we used, and resulted in higher hTIMP serum levels. 28 Adenoviruses have also been delivered directly into the tumor 24,27 or into normal, non-cancerous tissue to protect it against metastasis. 28 The main purpose of the study was to investigate whether constitutive expression of hTIMP gene products affected the outcome of TNF/IFNg antitumor therapy.…”

“…13 TIMPs have a pronounced antitumor potential, first observed with recombinant TIMP [14][15][16] and later with tumor cells engineered to overexpress TIMPs. [17][18][19][20][21] Recent data have shown that in vivo gene therapy with TIMPs is an excellent treatment for established primary tumors, at least in small animal models, [22][23][24][25][26][27] and also protects normal, non-cancerous target tissue against metastases. 28,29 The primary aim of this research was to examine the effects of constitutive expression of TIMP gene products on the outcome of antitumor therapy with TNF/IFNg.…”

Metalloproteinases and their inhibitors in tumor angiogenesis

“…Retrovirus-mediated expression of TIMP-3 in tumor cells inhib- 145 MMP-2 Various IFN-a 147 EC proliferation/migration Angiostatin 148,149 Various Endostatin 117,150 MT1-MMP, MMP-2 Various Arresten 151 EC proliferation, migration Canstatin 152 EC viability, migration Tumstatin 153,154 EC viability, migration, protein synthesis Platelet factor 4 155 VEGF and bFGF signaling TIMP-1 156 Broad-range Various effects, including MMP inhibition TIMP-2 157 Broad-range Various effects, including MMP inhibition TIMP-3 138 Broad-range Various effects, including MMP inhibition TIMP-4 136 Broad-range Various effects, including MMP inhibition Exogenous/synthetic TNP-470 158 EC proliferation Squalamine 159 EC proliferation Captopril 160 MMP-2, MMP-9 EC migration Combretastatin-A4 161 EC tubulin CP-547, 632 162 VEGFR-2 and bFGF kinases Vitaxin 163 a v b 3 integrin, EC apoptosis EMD121974 164 a v b 3 and a v b 5 integrin COL-3 83 MMP-2, -9, -14 MMPs Neovastat 165,166 MMP-2, -9, -12 MMPs, designed to avoid sheddases BMS-275291 54 MMP-1, -2, -3, -7, -9, -14 Bevacizumab 167 VEGF ited tumor growth and angiogenesis in vivo. Tumors overexpressing TIMP-3 failed to form a functional vasculature and had reduced pericyte recruitment.…”

Replicating retroviral vectors mediating continuous production and secretion of therapeutic gene products from cancer cells

“…Numerous experimental and clinical studies have indicated that the therapeutic efficacies of such treatment modalities essentially require continuous delivery of a large quantity of secretory therapeutic molecules. [13][14][15][16][17] Hence, gene delivery to endothelial cells should be able to allow boosting of the antiangiogenesis effect by their direct and durable transmission of the introduced therapeutic genes.…”

“…On the other hand, the capability of RCRs to integrate into host DNA makes them ideal candidates for long-term genetic modification of tumor cells which may be a considerable advantage for a number of genebased strategies, for example, anti-angiogenesis treatment or protease inhibitor-based therapy. [13][14][15][16][17] However, and despite these properties, the applicability of RCR vectors in cancer gene therapy has been limited, primarily due to the instability of replicating retroviruses carrying additional transgenes. [18][19][20] Previous studies have shown that replicating MLV carrying transgene sequences between 100 to 1200 bp within the U3 region of the 3 0 long terminal repeat (LTR) usually lost their inserts within a few passages.…”