The activity of a designer tissue inhibitor of metalloproteinases (TIMP)-1 against native membrane type 1 matrix metalloproteinase (MT1-MMP) in a cell-based environment

被引:20
作者
Lee, Meng-Huee [1 ]
Atkinson, Susan [1 ]
Rapti, Magdalini [1 ]
Handsley, Madeleine [2 ]
Curry, Valerie [1 ]
Edwards, Dylan [2 ]
Murphy, Gillian [1 ]
机构
[1] Univ Cambridge, Dept Oncol, Canc Res Inst, Li Ka Shing Ctr, Cambridge CB2 ORE, England
[2] Univ E Anglia, Sch Biol Sci, Norwich NR4 7TJ, Norfolk, England
基金
英国生物技术与生命科学研究理事会; 英国医学研究理事会;
关键词
TIMP-1; MT1-MMP; Site directed mutagenesis; Cell-based assays; Cancer therapeutics; ACTIVATION; PROGELATINASE; INVASION; CLEAVES; DOMAIN; TIMP-2; CD44; GELS;
D O I
10.1016/j.canlet.2009.08.029
中图分类号
R73 [肿瘤学];
学科分类号
100214 ;
摘要
The surface-anchored membrane type 1 matrix metalloproteinase (MT1-MMP) degrades a wide range of extracellular matrix components that includes collagens, laminins, fibronectin and the structural proteoglycan aggrecan. The enzyme modulates cell motility and plays an important role in tumour invasion and proliferation. We have previously designed a variant of tissue inhibitor of metalloproteinase (TIMP)-1 bearing a triple mutation (V4A + P6V + T98L, or N-TIMP-1(mt1)) that forms tight binary complex with the soluble catalytic domain of MT1-MMP [M.H. Lee, M. Rapti, G. Murphy, J. Biol. Chem. 278 (2003) 40224-40230]. Here, we report our latest findings on the cellular potency of this mutant against native MT1-MMP in cell-based environment. We show that N-TIMP-1(mt1) is a highly potent inhibitor against the ectodomain form of MT1-MMP (K-i 9.53 nM) with potential for further development as a therapeutic agent. The mutant is devoid of pro-MMP-2-activating capability but is highly effective in blocking MT1-MMP-mediated FITC-labelled collagen and gelatin film degradation in HTC75 fibrosarcoma and MCF7 breast cancer models. Most encouragingly, N-TIMP-1(mt1) is also effective against CD44 shedding in HTC75 cells and able to prevent tubule formation in human umbilical vascular endothelial cells (HUVEC) in a 3D fibrin gel model. We are interested in the development of the TIMPs as therapeutic agents against MT1-MMP related disorders such as cancers. Our findings here indicate the potential for the design of selective TIMPs with refined specificity and possibility for future therapeutic application. (C) 2009 Elsevier Ireland Ltd. All rights reserved.
引用
收藏
页码:114 / 122
页数:9
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