Tissue Inhibitor of Metalloproteinases-1 Interacts with CD74 to Promote AKT Signaling, Monocyte Recruitment Responses, and Vascular Smooth Muscle Cell Proliferation

Cells. 2023 Jul 20;12(14):1899. doi: 10.3390/cells12141899.

Abstract

Tissue inhibitor of metalloproteinases-1 (TIMP-1), an important regulator of matrix metalloproteinases (MMPs), has recently been shown to interact with CD74, a receptor for macrophage migration inhibitory factor (MIF). However, the biological effects mediated by TIMP-1 through CD74 remain largely unexplored. Using sequence alignment and in silico protein-protein docking analysis, we demonstrated that TIMP-1 shares residues with both MIF and MIF-2, crucial for CD74 binding, but not for CXCR4. Subcellular colocalization, immunoprecipitation, and internalization experiments supported these findings, demonstrating that TIMP-1 interacts with surface-expressed CD74, resulting in its internalization in a dose-dependent manner, as well as with a soluble CD74 ectodomain fragment (sCD74). This prompted us to study the effects of the TIMP-1-CD74 axis on monocytes and vascular smooth muscle cells (VSCMs) to assess its impact on vascular inflammation. A phospho-kinase array revealed the activation of serine/threonine kinases by TIMP-1 in THP-1 pre-monocytes, in particular AKT. Similarly, TIMP-1 dose-dependently triggered the phosphorylation of AKT and ERK1/2 in primary human monocytes. Importantly, Transwell migration, 3D-based Chemotaxis, and flow adhesion assays demonstrated that TIMP-1 engagement of CD74 strongly promotes the recruitment response of primary human monocytes, while live cell imaging studies revealed a profound activating effect on VSMC proliferation. Finally, re-analysis of scRNA-seq data highlighted the expression patterns of TIMP-1 and CD74 in human atherosclerotic lesions, thus, together with our experimental data, indicating a role for the TIMP-1-CD74 axis in vascular inflammation and atherosclerosis.

Keywords: atherogenesis; cell migration/adhesion; chemokine; cytokine; proliferation; vascular inflammation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Atherosclerosis*
  • Cell Proliferation
  • Humans
  • Inflammation
  • Monocytes*
  • Muscle, Smooth, Vascular
  • Proto-Oncogene Proteins c-akt
  • Tissue Inhibitor of Metalloproteinase-1

Substances

  • Proto-Oncogene Proteins c-akt
  • Tissue Inhibitor of Metalloproteinase-1

Grants and funding

This work was supported by Deutsche Forschungsgemeinschaft (DFG) Grant SFB1123-A3 to J.B. and A.K., SFB1123-A1/A10 to C.W., SFB-TRR57-A7 to J.B. and M.-L.B. (Project ID 36842431), DFG INST 409/209-1 FUGG to J.B., and by DFG under Germany’s Excellence Strategy within the framework of the Munich Cluster for Systems Neurology (EXC 2145 SyNergy—ID 390857198) to J.B.; J.M. received funding from the Dansk Kræftforskningsfond.