Tissue Inhibitor of Metalloproteinases-3 Peptides Inhibit Angiogenesis and Choroidal Neovascularization in Mice

Abstract

Tissue inhibitors of metalloproteinases (TIMPs) while originally characterized as inhibitors of matrix metalloproteinases (MMPs) have recently been shown to have a wide range of functions that are independent of their MMP inhibitory properties. Tissue inhibitor of metalloproteinases-3 (TIMP-3) is a potent inhibitor of VEGF-mediated angiogenesis and neovascularization through its ability to block the binding of VEGF to its receptor VEGFR-2. To identify and characterize the anti-angiogenic domain of TIMP-3, structure function analyses and synthetic peptide studies were performed using VEGF-mediated receptor binding, signaling, migration and proliferation. In addition, the ability of TIMP-3 peptides to inhibit CNV in a mouse model was evaluated. We demonstrate that the anti-angiogenic property resides in the COOH-terminal domain of TIMP-3 protein which can block the binding of VEGF specifically to its receptor VEGFR-2, but not to VEGFR-1 similar to the full-length wild-type protein. Synthetic peptides corresponding to putative loop 6 and tail region of TIMP-3 have anti-angiogenic properties as determined by inhibition of VEGF binding to VEGFR-2, VEGF-induced phosphorylation of VEGFR-2 and downstream signaling pathways as well as endothelial cell proliferation and migration in response to VEGF. In addition, we show that intravitreal administration of TIMP-3 peptide could inhibit the size of laser-induced choroidal neovascularization lesions in mice. Thus, we have identified TIMP-3 peptides to be efficient inhibitors of angiogenesis and have a potential to be used therapeutically in diseases with increased neovascularization.

Calcifying Cystic Odontogenic Tumour: immunohistochemical expression of matrix metalloproteinases, their inhibitors (TIMPs and RECK) and inducer (EMMPRIN)

“…This suggests that TIMP‐3 is a major regulator of MMP activities in vivo . Furthermore, TIMP‐3 plays a key role in innate immunity by regulating the processing of tumour necrosis factor‐α (TNF‐α) by ADAM17 is implicated in vascular inhibition by blockage of VEGF binding to its receptor , by interaction to angiotensin II type 2 , as well as TIMP‐3 knockout mice have been showed neovascularization . Although TIMP‐3 was strongly detected in both neoplastic odontogenic epithelium and adjacent stroma of CCOT, nothing is known in the literature about its expression in odontogenic tumours.…”

Tissue inhibitor of metalloproteinase-3 (TIMP3) promotes endothelial apoptosis via a caspase-independent mechanism

“…In vitro formation of tubular structures was studied with a modified three-dimensional matrix assay as previous described (27, 34, 35). Briefly, the cells pretreated with or without caspase inhibitors for 30 min were subjected to a first layer of growth factor-reduced Matrigel (Becton Dickinson, Bedford, MA) in 24 well plates at 2×10 4 cells/well.…”

Molecular characterization and hormonal regulation of tissue inhibitor of metalloproteinase 1 in goat ovarian granulosa cells

“…TIMP proteins possess a similar domain structure, which is composed of an Nterminal domain and a C-terminal domain [4]. Previous studies have shown that the members of TIMPs have key functions in various physiological processes, including morphogenesis, reproduction, cancer, arthritis, and angiogenesis [5][6][7][8][9][10]. TIMP1 is the first member of the TIMPs family with multiple functions in numerous human and rodent cells [4,11].…”