A top-down approach to uncover the hidden ligandome of low-density lipoprotein receptor-related protein 1 in cartilage

Kazuhiro Yamamoto; Carsten Scavenius; Maria M. Meschis; Abdulrahman M.E. Gremida; Emilie H. Mogensen; Ida B. Thøgersen; Simone Bonelli; Simone D. Scilabra; Anders Jensen; Salvatore Santamaria; Josefin Ahnström; George Bou-Gharios; Jan J. Enghild; Hideaki Nagase

doi:10.1016/j.matbio.2022.08.007

A top-down approach to uncover the hidden ligandome of low-density lipoprotein receptor-related protein 1 in cartilage

Kazuhiro Yamamoto^*, Carsten Scavenius, Maria M. Meschis, Abdulrahman M.E. Gremida, Emilie H. Mogensen, Ida B. Thøgersen, Simone Bonelli, Simone D. Scilabra, Anders Jensen, Salvatore Santamaria, Josefin Ahnström, George Bou-Gharios, Jan J. Enghild, Hideaki Nagase

^*Corresponding author af dette arbejde

Institut for Molekylærbiologi og Genetik - Proteinvidenskab

Publikation: Bidrag til tidsskrift/Konferencebidrag i tidsskrift /Bidrag til avis › Tidsskriftartikel › Forskning › peer review

10 Citationer (Scopus)

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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.

Originalsprog	Engelsk
Tidsskrift	Matrix Biology
Vol/bind	112
Sider (fra-til)	190-218
Antal sider	29
ISSN	0945-053X
DOI	https://doi.org/10.1016/j.matbio.2022.08.007
Status	Udgivet - sep. 2022

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10.1016/j.matbio.2022.08.007Licens: CC BY

1-s2.0-S0945053X22001019-mainForlagets udgivne version, 4,43 MBLicens: CC BY

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Yamamoto, K., Scavenius, C., Meschis, M. M., Gremida, A. M. E., Mogensen, E. H., Thøgersen, I. B., Bonelli, S., Scilabra, S. D., Jensen, A., Santamaria, S., Ahnström, J., Bou-Gharios, G., Enghild, J. J., & Nagase, H. (2022). A top-down approach to uncover the hidden ligandome of low-density lipoprotein receptor-related protein 1 in cartilage. Matrix Biology, 112, 190-218. https://doi.org/10.1016/j.matbio.2022.08.007

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title = "A top-down approach to uncover the hidden ligandome of low-density lipoprotein receptor-related protein 1 in cartilage",

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.",

keywords = "Cartilage, Chondrocyte, Endocytosis, Extracellular matrix, Interactome, LRP1, Osteoarthritis",

author = "Kazuhiro Yamamoto and Carsten Scavenius and Meschis, {Maria M.} and Gremida, {Abdulrahman M.E.} and Mogensen, {Emilie H.} and Th{\o}gersen, {Ida B.} and Simone Bonelli and Scilabra, {Simone D.} and Anders Jensen and Salvatore Santamaria and Josefin Ahnstr{\"o}m and George Bou-Gharios and Enghild, {Jan J.} and Hideaki Nagase",

note = "Publisher Copyright: {\textcopyright} 2022",

year = "2022",

month = sep,

doi = "10.1016/j.matbio.2022.08.007",

language = "English",

volume = "112",

pages = "190--218",

journal = "Matrix Biology",

issn = "0945-053X",

publisher = "Elsevier B.V.",

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Yamamoto, K, Scavenius, C, Meschis, MM, Gremida, AME, Mogensen, EH, Thøgersen, IB, Bonelli, S, Scilabra, SD, Jensen, A, Santamaria, S, Ahnström, J, Bou-Gharios, G, Enghild, JJ & Nagase, H 2022, 'A top-down approach to uncover the hidden ligandome of low-density lipoprotein receptor-related protein 1 in cartilage', Matrix Biology, bind 112, s. 190-218. https://doi.org/10.1016/j.matbio.2022.08.007

TY - JOUR

T1 - A top-down approach to uncover the hidden ligandome of low-density lipoprotein receptor-related protein 1 in cartilage

AU - Yamamoto, Kazuhiro

AU - Scavenius, Carsten

AU - Meschis, Maria M.

AU - Gremida, Abdulrahman M.E.

AU - Mogensen, Emilie H.

AU - Thøgersen, Ida B.

AU - Bonelli, Simone

AU - Scilabra, Simone D.

AU - Jensen, Anders

AU - Santamaria, Salvatore

AU - Ahnström, Josefin

AU - Bou-Gharios, George

AU - Enghild, Jan J.

AU - Nagase, Hideaki

PY - 2022/9

Y1 - 2022/9

N2 - 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.

AB - 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.

KW - Cartilage

KW - Chondrocyte

KW - Endocytosis

KW - Extracellular matrix

KW - Interactome

KW - LRP1

KW - Osteoarthritis

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U2 - 10.1016/j.matbio.2022.08.007

DO - 10.1016/j.matbio.2022.08.007

M3 - Journal article

C2 - 36028175

AN - SCOPUS:85137616621

SN - 0945-053X

VL - 112

SP - 190

EP - 218

JO - Matrix Biology

JF - Matrix Biology

ER -

A top-down approach to uncover the hidden ligandome of low-density lipoprotein receptor-related protein 1 in cartilage

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