Yamamoto, Kazuhiro 
ORCID: 0000-0002-8481-775X, Scavenius, Carsten, Meschis, Maria M, Gremida, Abdulrahman ME, Mogensen, Emilie H, Thøgersen, Ida B, Bonelli, Simone, Scilabra, Simone D, Jensen, Anders, Santamaria, Salvatore et al (show 4 more authors)
(2022)
A top-down approach to uncover the hidden ligandome of low-density lipoprotein receptor-related protein 1 in cartilage.
Matrix biology : journal of the International Society for Matrix Biology, 112.
S0945-053X(22)00101-9-S0945-053X(22)00101-9.
Abstract
The low-density lipoprotein receptor-related protein 1 (LRP1) is a cell-surface receptor ubiquitously expressed in various tissues. It plays tissue-specific roles by mediating endocytosis of a diverse range of extracellular molecules. Dysregulation of LRP1 is involved in multiple conditions including osteoarthritis (OA) but little information is available about the specific profile of direct binding partners of LRP1 (ligandome) for each tissue, which would lead to a better understanding of its role in disease states. Here, we investigated adult articular cartilage where impaired LRP1-mediated endocytosis leads to tissue destruction. We used a top-down approach involving proteomic analysis of the LRP1 interactome in human chondrocytes, direct binding assays using purified LRP1 and ligand candidates, and validation in LRP1-deficient fibroblasts and human chondrocytes, as well as a novel Lrp1 conditional knockout (KO) mouse model. We found that inhibition of LRP1 and ligand interaction results in cell death, alteration of the entire secretome and transcriptional modulations in human chondrocytes. We identified a chondrocyte-specific LRP1 ligandome consisting of more than 50 novel ligand candidates. Surprisingly, 23 previously reported LRP1 ligands were not regulated by LRP1-mediated endocytosis in human chondrocytes. We confirmed direct LRP1 binding of HGFAC, HMGB1, HMGB2, CEMIP, SLIT2, ADAMTS1, TSG6, IGFBP7, SPARC and LIF, correlation between their affinity for LRP1 and the rate of endocytosis, and some of their intracellular localization. Moreover, a conditional LRP1 KO mouse model demonstrated a critical role of LRP1 in regulating the high-affinity ligands in cartilage in vivo. This systematic approach revealed the specificity and the extent of the chondrocyte LRP1 ligandome and identified potential novel therapeutic targets for OA.
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | Cartilage, Articular, Animals, Mice, Knockout, Humans, Mice, Osteoarthritis, Lipoproteins, LDL, HMGB1 Protein, HMGB2 Protein, Ligands, Proteomics, Adult, Low Density Lipoprotein Receptor-Related Protein-1 |
| Divisions: | Faculty of Health and Life Sciences Faculty of Health and Life Sciences > Institute of Life Courses and Medical Sciences |
| Depositing User: | Symplectic Admin |
| Date Deposited: | 05 Sep 2022 07:32 |
| Last Modified: | 18 Jan 2023 20:45 |
| DOI: | 10.1016/j.matbio.2022.08.007 |
| Open Access URL: | https://authors.elsevier.com/sd/article/S0945-053X... |
| Related URLs: | |
| URI: | https://livrepository.liverpool.ac.uk/id/eprint/3163276 |
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