Retroviral vector-producer cell-mediated in vivo gene transfer of TIMP-3 restricts angiogenesis and neuroblastoma growth in mice

Abstract

Destruction and remodeling of the extracellular matrix occurs during the formation of new blood vessels that are required for tumor growth. We sought to determine whether gene-therapy mediated in vivo delivery of tissue inhibitor of matrix metalloproteinase-3 (TIMP-3), using retroviral vector-producer cells, could suppress angiogenesis and subsequent tumor growth in a murine neuroblastoma model. Tumor volume 28 days after coinjection of tumor cells with producer cells generating TIMP-3-encoding retroviral vectors was 21% that of controls, as was the mean tumor vascular index, a measure of blood vessel maturity. When tumors were allowed to reach a mean volume of 0.05 cm 3 before treatment, their size 2 weeks later was 47% relative to controls; larger tumors were not significantly affected. When producer cells were injected at surgical sites following excision of subcutaneous tumors, local recurrence 14 days later was only 22% in TIMP-3 producer cell treated mice as compared to 71% in controls. Unsuccessful transduction of melanoma cells in situ, another tumor of neural crest origin, resulted in unimpaired tumor growth, despite the fact that these tumors are susceptible to TIMP-3 overexpression, demonstrating the importance of tumor cell transduction in this approach. Thus, retroviral vector-producer cell-mediated in vivo gene transfer of TIMP-3 to tumor cells can significantly restrict tumor-induced angiogenesis and tumor growth. This approach may be an effective adjuvant in the treatment of neuroblastoma and other solid tumors refractory to traditional therapy, although it appears to be most effective in smaller tumors or in the setting of minimal residual disease, and the tumor cells must be susceptible to retroviral vector-mediated transduction.

Inhibition of tumor growth and induction of apoptosis in prostate cancer cell lines by overexpression of tissue inhibitor of matrix metalloproteinase-3

“…Western blot and immunohistochemical assays showed that TIMP-3 is expressed at low levels both in human primary prostate cancer tissues and in prostate cancer cell lines. These data are consistent with other studies 7,11,21 that have shown loss or low expression of TIMP-3 in a variety of human tumors including prostate cancer. RT-PCR and western blot analyses showed that TIMP-3 is expressed at low levels in prostate cancer cell line PC-3M.…”

“…The mechanism may be mediated through inhibition of functional capillary morphogenesis. 21 Measurements of tumorigenicity in vivo study showed that TIMP-3 overexpressing cells completely lost the ability to form tumors. This finding is similar with the result of Bian et al 25 which showed that although TIMP-3 overexpressing cells can grow in soft agar, they failed to grow in nude mice, and Lam et al 44 who showed growth inhibition of human glioma tumor xenografts in immunodeficient mice.…”

Membrane type-matrix metalloproteinases and tumor progression

“…TIMP-1 has no effect on the VEGF levels in both MT1 -MMP transfected cells, however, TIMP-1 can up-regulate VEGF in MCF7 [169]. Overexpression of TIMP-3 in neuroblastoma and melanoma by retroviral delivery decreases tumor angiogenesis in severe combined immunodeficient (SCID) mice by inhibition of functional capillary morphogenesis [170,171]. Human tissue factor pathway inhibitor-2 (TFPI-2) that inhibits the plasmin and trypsin-mediated activation of proMMPs has been reported to inhibit in vivo growth and metastasis of highly metastatic fibrosarcoma cells (HT1080) through a downregulation of VEGF expression in cells overexpressing TFPI-2 cells [172].…”

Tissue Inhibitors of Metalloproteinase 2 Inhibits Endothelial Cell Migration through Increased Expression of RECK

“…TIMP-3 inhibits capillary morphogenesis in vivo and endothelial cell migration in vitro, both effects reportedly mediated by inhibition of MMP activity. However, TIMP-3 has been shown recently to function as an antagonist for the vascular endothelial growth factor receptor (VEGFR)-2, inhibiting vascular endothelial growth factor (VEGF)-A binding to this receptor (17,18). It has also been reported that TIMP-2 inhibits cell migration after TIMP-2 transfection of human microvascular endothelial cells (hMVECs; ref.…”