Targeting a Designer TIMP-1 to the Cell Surface for Effective MT1-MMP Inhibition: A Potential Role for the Prion Protein in Renal Carcinoma Therapy

Jiang, Bingjie, Liu, Jian and Lee, Meng Huee (2019) Targeting a Designer TIMP-1 to the Cell Surface for Effective MT1-MMP Inhibition: A Potential Role for the Prion Protein in Renal Carcinoma Therapy. MOLECULES, 24 (2). E255-.

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Abstract

Renal carcinoma cells express Membrane Type 1-Matrix Metalloproteinase (MT1-MMP, MMP-14) to degrade extracellular matrix components and a range of bioactive molecules to allow metastasis and cell proliferation. The activity of MT1-MMP is modulated by the endogenous inhibitors, Tissue Inhibitor of Metalloproteinases (TIMPs). In this study, we describe a novel strategy that would enable a "designer" TIMP-1 tailored specifically for MT1-MMP inhibition (V4A/P6V/T98L; <i>K</i>_i^app 1.66 nM) to be targeted to the plasma membrane for more effective MT1-MMP inhibition. To achieve this, we fuse the designer TIMP-1 to the glycosyl-phosphatidyl inositol (GPI) anchor of the prion protein to create a membrane-tethered, high-affinity TIMP variant named "T1^{Pr αMT1}" that is predominantly located on the cell surface and co-localised with MT1-MMP. Confocal microscopy shows that T1^{Pr αMT1} is found throughout the cell surface in particular the membrane ruffles where MT1-MMP is most abundant. Expression of T1^{Pr αMT1} brings about a complete abrogation of the gelatinolytic activity of cellular MT1-MMP in HT1080 fibrosarcoma cells whilst in renal carcinoma cells CaKi-1, the GPI-TIMP causes a disruption in MMP-mediated proteolysis of ECM components such as fibronectin, collagen I and laminin that consequently triggers a downstream senescence response. Moreover, the transduced cells also suffer from an impairment in proliferation and survival in vitro as well as in NOD/SCID mouse xenograft. Taken together, our findings demonstrate that the GPI anchor of prion could be exploited as a targeting device in TIMP engineering for MT1-MMP inhibition with a potential in renal carcinoma therapy.

Item Type:	Article
Uncontrolled Keywords:	MT1-MMP, TIMP, renal carcinoma, cancer therapy, prion, GPI anchor, protein engineering
Depositing User:	Symplectic Admin
Date Deposited:	21 Mar 2019 12:01
Last Modified:	07 Feb 2024 17:57
DOI:	10.3390/molecules24020255
Related URLs:	Author Publisher
URI:	https://livrepository.liverpool.ac.uk/id/eprint/3034601