Receptor Protein Tyrosine Phosphatase α-Mediated Enhancement of Rheumatoid Synovial Fibroblast Signaling and Promotion of Arthritis in Mice

Stephanie M Stanford; Mattias N D Svensson; Cristiano Sacchetti; Caila A Pilo; Dennis J Wu; William B Kiosses; Annelie Hellvard; Brith Bergum; German R Aleman Muench; Christian Elly; Yun-Cai Liu; Jeroen den Hertog; Ari Elson; Jan Sap; Piotr Mydel; David L Boyle; Maripat Corr; Gary S Firestein; Nunzio Bottini

doi:10.1002/art.39442

Receptor Protein Tyrosine Phosphatase α-Mediated Enhancement of Rheumatoid Synovial Fibroblast Signaling and Promotion of Arthritis in Mice

Stephanie M Stanford
, Mattias N D Svensson
, Cristiano Sacchetti
, Caila A Pilo
, Dennis J Wu
, William B Kiosses
, Annelie Hellvard
, Brith Bergum
, German R Aleman Muench
, Christian Elly
, Yun-Cai Liu
, Jeroen den Hertog
, Ari Elson
, Jan Sap
, Piotr Mydel
, David L Boyle
, Maripat Corr
, Gary S Firestein
, Nunzio Bottini

Cancer

Research output: Contribution to journal › Article › Academic › peer-review

Abstract

OBJECTIVE: During rheumatoid arthritis (RA), fibroblast-like synoviocytes (FLS) critically promote disease pathogenesis by aggressively invading the extracellular matrix of the joint. The focal adhesion kinase (FAK) signaling pathway is emerging as a contributor to the anomalous behavior of RA FLS. The receptor protein tyrosine phosphatase α (RPTPα), which is encoded by the PTPRA gene, is a key promoter of FAK signaling. The aim of this study was to investigate whether RPTPα mediates FLS aggressiveness and RA pathogenesis.

METHODS: Through RPTPα knockdown, we assessed FLS gene expression by quantitative polymerase chain reaction analysis and enzyme-linked immunosorbent assay, invasion and migration by Transwell assays, survival by annexin V and propidium iodide staining, adhesion and spreading by immunofluorescence microscopy, and activation of signaling pathways by Western blotting of FLS lysates. Arthritis development was examined in RPTPα-knockout (KO) mice using the K/BxN serum-transfer model. The contribution of radiosensitive and radioresistant cells to disease was evaluated by reciprocal bone marrow transplantation.

RESULTS: RPTPα was enriched in the RA synovial lining. RPTPα knockdown impaired RA FLS survival, spreading, migration, invasiveness, and responsiveness to platelet-derived growth factor, tumor necrosis factor, and interleukin-1 stimulation. These phenotypes correlated with increased phosphorylation of Src on inhibitory Y(527) and decreased phosphorylation of FAK on stimulatory Y(397) . Treatment of RA FLS with an inhibitor of FAK phenocopied the knockdown of RPTPα. RPTPα-KO mice were protected from arthritis development, which was due to radioresistant cells.

CONCLUSION: By regulating the phosphorylation of Src and FAK, RPTPα mediates proinflammatory and proinvasive signaling in RA FLS, correlating with the promotion of disease in an FLS-dependent model of RA.

Original language	English
Pages (from-to)	359-369
Number of pages	11
Journal	Arthritis & Rheumatology
Volume	68
Issue number	2
DOIs	https://doi.org/10.1002/art.39442
Publication status	Published - Feb 2016

Keywords

Animals
Ankle Joint
Apoptosis
Arthritis, Experimental
Arthritis, Rheumatoid
Blotting, Western
Cell Adhesion
Cell Movement
Cell Survival
Disease Progression
Enzyme-Linked Immunosorbent Assay
Fibroblasts
Focal Adhesion Protein-Tyrosine Kinases
Gene Expression Profiling
Gene Knockdown Techniques
Interleukin-1
Mice
Mice, Knockout
Phosphorylation
Platelet-Derived Growth Factor
Polymerase Chain Reaction
Receptor-Like Protein Tyrosine Phosphatases, Class 4
Signal Transduction
Synovial Membrane
Tumor Necrosis Factor-alpha
src-Family Kinases

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Stanford, S. M., Svensson, M. N. D., Sacchetti, C., Pilo, C. A., Wu, D. J., Kiosses, W. B., Hellvard, A., Bergum, B., Muench, G. R. A., Elly, C., Liu, Y.-C., den Hertog, J., Elson, A., Sap, J., Mydel, P., Boyle, D. L., Corr, M., Firestein, G. S., & Bottini, N. (2016). Receptor Protein Tyrosine Phosphatase α-Mediated Enhancement of Rheumatoid Synovial Fibroblast Signaling and Promotion of Arthritis in Mice. Arthritis & Rheumatology, 68(2), 359-369. https://doi.org/10.1002/art.39442

@article{c178a6ed00a24f2ab79b5cc059d2ae30,

title = "Receptor Protein Tyrosine Phosphatase α-Mediated Enhancement of Rheumatoid Synovial Fibroblast Signaling and Promotion of Arthritis in Mice",

abstract = "OBJECTIVE: During rheumatoid arthritis (RA), fibroblast-like synoviocytes (FLS) critically promote disease pathogenesis by aggressively invading the extracellular matrix of the joint. The focal adhesion kinase (FAK) signaling pathway is emerging as a contributor to the anomalous behavior of RA FLS. The receptor protein tyrosine phosphatase α (RPTPα), which is encoded by the PTPRA gene, is a key promoter of FAK signaling. The aim of this study was to investigate whether RPTPα mediates FLS aggressiveness and RA pathogenesis.METHODS: Through RPTPα knockdown, we assessed FLS gene expression by quantitative polymerase chain reaction analysis and enzyme-linked immunosorbent assay, invasion and migration by Transwell assays, survival by annexin V and propidium iodide staining, adhesion and spreading by immunofluorescence microscopy, and activation of signaling pathways by Western blotting of FLS lysates. Arthritis development was examined in RPTPα-knockout (KO) mice using the K/BxN serum-transfer model. The contribution of radiosensitive and radioresistant cells to disease was evaluated by reciprocal bone marrow transplantation.RESULTS: RPTPα was enriched in the RA synovial lining. RPTPα knockdown impaired RA FLS survival, spreading, migration, invasiveness, and responsiveness to platelet-derived growth factor, tumor necrosis factor, and interleukin-1 stimulation. These phenotypes correlated with increased phosphorylation of Src on inhibitory Y(527) and decreased phosphorylation of FAK on stimulatory Y(397) . Treatment of RA FLS with an inhibitor of FAK phenocopied the knockdown of RPTPα. RPTPα-KO mice were protected from arthritis development, which was due to radioresistant cells.CONCLUSION: By regulating the phosphorylation of Src and FAK, RPTPα mediates proinflammatory and proinvasive signaling in RA FLS, correlating with the promotion of disease in an FLS-dependent model of RA.",

keywords = "Animals, Ankle Joint, Apoptosis, Arthritis, Experimental, Arthritis, Rheumatoid, Blotting, Western, Cell Adhesion, Cell Movement, Cell Survival, Disease Progression, Enzyme-Linked Immunosorbent Assay, Fibroblasts, Focal Adhesion Protein-Tyrosine Kinases, Gene Expression Profiling, Gene Knockdown Techniques, Interleukin-1, Mice, Mice, Knockout, Phosphorylation, Platelet-Derived Growth Factor, Polymerase Chain Reaction, Receptor-Like Protein Tyrosine Phosphatases, Class 4, Signal Transduction, Synovial Membrane, Tumor Necrosis Factor-alpha, src-Family Kinases",

author = "Stanford, \{Stephanie M\} and Svensson, \{Mattias N D\} and Cristiano Sacchetti and Pilo, \{Caila A\} and Wu, \{Dennis J\} and Kiosses, \{William B\} and Annelie Hellvard and Brith Bergum and Muench, \{German R Aleman\} and Christian Elly and Yun-Cai Liu and \{den Hertog\}, Jeroen and Ari Elson and Jan Sap and Piotr Mydel and Boyle, \{David L\} and Maripat Corr and Firestein, \{Gary S\} and Nunzio Bottini",

note = "{\textcopyright} 2016, American College of Rheumatology.",

year = "2016",

month = feb,

doi = "10.1002/art.39442",

language = "English",

volume = "68",

pages = "359--369",

journal = "Arthritis \& Rheumatology",

issn = "2326-5191",

publisher = "John Wiley \& Sons Inc.",

number = "2",

}

Stanford, SM, Svensson, MND, Sacchetti, C, Pilo, CA, Wu, DJ, Kiosses, WB, Hellvard, A, Bergum, B, Muench, GRA, Elly, C, Liu, Y-C, den Hertog, J, Elson, A, Sap, J, Mydel, P, Boyle, DL, Corr, M, Firestein, GS & Bottini, N 2016, 'Receptor Protein Tyrosine Phosphatase α-Mediated Enhancement of Rheumatoid Synovial Fibroblast Signaling and Promotion of Arthritis in Mice', Arthritis & Rheumatology, vol. 68, no. 2, pp. 359-369. https://doi.org/10.1002/art.39442

TY - JOUR

T1 - Receptor Protein Tyrosine Phosphatase α-Mediated Enhancement of Rheumatoid Synovial Fibroblast Signaling and Promotion of Arthritis in Mice

AU - Stanford, Stephanie M

AU - Svensson, Mattias N D

AU - Sacchetti, Cristiano

AU - Pilo, Caila A

AU - Wu, Dennis J

AU - Kiosses, William B

AU - Hellvard, Annelie

AU - Bergum, Brith

AU - Muench, German R Aleman

AU - Elly, Christian

AU - Liu, Yun-Cai

AU - den Hertog, Jeroen

AU - Elson, Ari

AU - Sap, Jan

AU - Mydel, Piotr

AU - Boyle, David L

AU - Corr, Maripat

AU - Firestein, Gary S

AU - Bottini, Nunzio

PY - 2016/2

Y1 - 2016/2

N2 - OBJECTIVE: During rheumatoid arthritis (RA), fibroblast-like synoviocytes (FLS) critically promote disease pathogenesis by aggressively invading the extracellular matrix of the joint. The focal adhesion kinase (FAK) signaling pathway is emerging as a contributor to the anomalous behavior of RA FLS. The receptor protein tyrosine phosphatase α (RPTPα), which is encoded by the PTPRA gene, is a key promoter of FAK signaling. The aim of this study was to investigate whether RPTPα mediates FLS aggressiveness and RA pathogenesis.METHODS: Through RPTPα knockdown, we assessed FLS gene expression by quantitative polymerase chain reaction analysis and enzyme-linked immunosorbent assay, invasion and migration by Transwell assays, survival by annexin V and propidium iodide staining, adhesion and spreading by immunofluorescence microscopy, and activation of signaling pathways by Western blotting of FLS lysates. Arthritis development was examined in RPTPα-knockout (KO) mice using the K/BxN serum-transfer model. The contribution of radiosensitive and radioresistant cells to disease was evaluated by reciprocal bone marrow transplantation.RESULTS: RPTPα was enriched in the RA synovial lining. RPTPα knockdown impaired RA FLS survival, spreading, migration, invasiveness, and responsiveness to platelet-derived growth factor, tumor necrosis factor, and interleukin-1 stimulation. These phenotypes correlated with increased phosphorylation of Src on inhibitory Y(527) and decreased phosphorylation of FAK on stimulatory Y(397) . Treatment of RA FLS with an inhibitor of FAK phenocopied the knockdown of RPTPα. RPTPα-KO mice were protected from arthritis development, which was due to radioresistant cells.CONCLUSION: By regulating the phosphorylation of Src and FAK, RPTPα mediates proinflammatory and proinvasive signaling in RA FLS, correlating with the promotion of disease in an FLS-dependent model of RA.

AB - OBJECTIVE: During rheumatoid arthritis (RA), fibroblast-like synoviocytes (FLS) critically promote disease pathogenesis by aggressively invading the extracellular matrix of the joint. The focal adhesion kinase (FAK) signaling pathway is emerging as a contributor to the anomalous behavior of RA FLS. The receptor protein tyrosine phosphatase α (RPTPα), which is encoded by the PTPRA gene, is a key promoter of FAK signaling. The aim of this study was to investigate whether RPTPα mediates FLS aggressiveness and RA pathogenesis.METHODS: Through RPTPα knockdown, we assessed FLS gene expression by quantitative polymerase chain reaction analysis and enzyme-linked immunosorbent assay, invasion and migration by Transwell assays, survival by annexin V and propidium iodide staining, adhesion and spreading by immunofluorescence microscopy, and activation of signaling pathways by Western blotting of FLS lysates. Arthritis development was examined in RPTPα-knockout (KO) mice using the K/BxN serum-transfer model. The contribution of radiosensitive and radioresistant cells to disease was evaluated by reciprocal bone marrow transplantation.RESULTS: RPTPα was enriched in the RA synovial lining. RPTPα knockdown impaired RA FLS survival, spreading, migration, invasiveness, and responsiveness to platelet-derived growth factor, tumor necrosis factor, and interleukin-1 stimulation. These phenotypes correlated with increased phosphorylation of Src on inhibitory Y(527) and decreased phosphorylation of FAK on stimulatory Y(397) . Treatment of RA FLS with an inhibitor of FAK phenocopied the knockdown of RPTPα. RPTPα-KO mice were protected from arthritis development, which was due to radioresistant cells.CONCLUSION: By regulating the phosphorylation of Src and FAK, RPTPα mediates proinflammatory and proinvasive signaling in RA FLS, correlating with the promotion of disease in an FLS-dependent model of RA.

KW - Animals

KW - Ankle Joint

KW - Apoptosis

KW - Arthritis, Experimental

KW - Arthritis, Rheumatoid

KW - Blotting, Western

KW - Cell Adhesion

KW - Cell Movement

KW - Cell Survival

KW - Disease Progression

KW - Enzyme-Linked Immunosorbent Assay

KW - Fibroblasts

KW - Focal Adhesion Protein-Tyrosine Kinases

KW - Gene Expression Profiling

KW - Gene Knockdown Techniques

KW - Interleukin-1

KW - Mice

KW - Mice, Knockout

KW - Phosphorylation

KW - Platelet-Derived Growth Factor

KW - Polymerase Chain Reaction

KW - Receptor-Like Protein Tyrosine Phosphatases, Class 4

KW - Signal Transduction

KW - Synovial Membrane

KW - Tumor Necrosis Factor-alpha

KW - src-Family Kinases

U2 - 10.1002/art.39442

DO - 10.1002/art.39442

M3 - Article

C2 - 26414708

SN - 2326-5191

VL - 68

SP - 359

EP - 369

JO - Arthritis & Rheumatology

JF - Arthritis & Rheumatology

IS - 2

ER -

Receptor Protein Tyrosine Phosphatase α-Mediated Enhancement of Rheumatoid Synovial Fibroblast Signaling and Promotion of Arthritis in Mice

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Keywords

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