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.

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; Kiapp 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 "T1Pr αMT1" that is predominantly located on the cell surface and co-localised with MT1-MMP. Confocal microscopy shows that T1Pr αMT1 is found throughout the cell surface in particular the membrane ruffles where MT1-MMP is most abundant. Expression of T1Pr 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.

• Authors: Bingjie Jiang, Jian Liu, Meng Huee Lee
• Journal: Molecules (Basel, Switzerland) (Molecules) Vol. 24 Issue 2 (Jan 11 2019) ISSN: 1420-3049 [Electronic] Switzerland
• PMID: 30641935 (Publication Type: Journal Article)

Chemical References

• Matrix Metalloproteinase Inhibitors
• Prion Proteins
• Tissue Inhibitor of Metalloproteinase-1
• Matrix Metalloproteinase 14

Topics

• Animals
• Carcinoma, Renal Cell (drug therapy, genetics, metabolism, pathology)
• Cell Line, Tumor
• Cell Membrane (metabolism)
• Cellular Senescence
• Disease Models, Animal
• Gene Expression
• Humans
• Matrix Metalloproteinase 14 (chemistry, genetics, metabolism)
• Matrix Metalloproteinase Inhibitors (chemistry, pharmacology)
• Mice
• Prion Proteins (antagonists & inhibitors)
• Tissue Inhibitor of Metalloproteinase-1 (antagonists & inhibitors, chemistry, metabolism)
• Xenograft Model Antitumor Assays