Investigation of the vesicle transport of MT1-MMP

Project No: KTPS-NC-5
Intake: 2021 KTPS-NC

PROJECT OVERVIEW

When cells migrate in tissue, the surrounding extracellular matrix (ECM) is a crucial scaffolding. However, it is also a physical barrier to prevent cell migration, which needs to be degraded locally at the leading edge to make a path for migration. Cells use a proteolytic enzyme called membrane-type 1 matrix metalloproteinase (MT1-MMP) to degrade ECM. MT1-MMP is a type I transmembrane proteinase expressed in various cell types with invasive character, including cancer cells, endothelial cells, fibroblasts, and myeloid cells. MT1-MMP directly degrades various ECM components on the cell surface, cleaves cell surface molecules, including cell adhesion molecules and receptors. It also exerts non-proteolytic functions such as the Warburg effect through the cytoplasmic tail. All of these activities have been shown to enhance cellular motility and invasion. One of the crucial regulatory mechanisms of MT1-MMP in invading cells is the dynamic localisation of MT1-MMP at the leading edges where it functions. It has been shown that this is achieved by a targeted transport of MT1-MMP-containing vesicles along microtubules. We have recently identified three kinesin motor proteins that are involved in MT1-MMP-containing vesicle transport. Among them, two of them collaborate to carry the same vesicle to the leading edge, while one of them carries the vesicle to other areas of the plasma membrane. It is not expected that these kinesin motor proteins directly interact with the enzyme to recognise MT1-MMP-containing vesicles, but adaptor molecules should mediate their interactions. In order to understand the mechanism of MT1-MMP vesicle transport, it is crucial to identify these adaptor molecules and signalling molecules that regulate vesicle transport. In this DPhil project, we will identify the adaptor molecules and its regulatory molecules that are the vicinity of MT1-MMP and the kinesin motor protein using proximity labelling followed by proteomics analyses. Upon identifying candidate molecules, we will investigate their function in regulating MT1-MMP vesicle transport by live-cell imaging using our state of art microscopies. Achieving this project will reveal a novel cellular invasion mechanism, contributing to cancer progression and various inflammatory diseases.

KEYWORDS

MT1-MMP, Kinesin motor proteins, proximity labelling, invasion

The Kennedy Institute is a world-renowned research centre and is housed in a state-of-the-art research facility. Full training will be provided in a range of cell and molecular biology techniques. A core curriculum of 20 lectures will be taken in the first term of year 1 to provide a solid foundation in musculoskeletal sciences, immunology and data analysis. Students will attend weekly departmental meetings and will be expected to attend seminars within the department and those relevant in the wider University. Subject-specific training will be received through our group's weekly supervision meetings. Students will also attend external scientific conferences where they will be expected to present the research findings. Achieving this project will provide a significant information

KEY PUBLICATIONS

Gifford V and Itoh Y (2019) MT1-MMP-dependent cell migration: proteolytic and non-proteolytic mechanisms. Biochem Soc Trans, DOI: 10.1042/BST20180363
Søgaard PP, Ito N, Sato N, Fujita, Matter K, and Itoh Y. (2019) Epithelial polarization in 3D matrix requires DDR1 signaling to regulate actomyosin contractility. Life Science Alliance, 2, e201800276. DOI: 10.26508/lsa.201800276
Majkowska I, Shitomi Y, Ito N, Gray NS, Itoh Y (2017) Discoidin Domain Receptor 2 Mediates Collagen-Induced Activation of Membrane-Type 1 Matrix Metalloproteinase in Human Fibroblasts. J Biol Chem, 292(16):6633-6643
Weaver SA, Wolters B, Ito N, Woskowicz AM, Kaneko K, Shitomi Y, Seiki M, Itoh Y. (2014) Basal localization of MT1-MMP is essential for epithelial cell morphogenesis in 3D collagen matrix. J Cell Sci. 127, 1203-13.
Woskowicz AM, Weaver SA, Shitomi Y, Ito N, Itoh Y. (2013) MT-LOOP-dependent localization of MT1-MMP to the cell adhesion complexes promotes cancer cell invasion. J Biol Chem. 288, 35126-37.