Receptor protein tyrosine phosphatase alpha enhances rheumatoid synovial fibroblast signaling and promotes 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 alpha enhances rheumatoid synovial fibroblast signaling and promotes 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

Hubrecht Institute

Research output: Contribution to journal/periodical › Article › Scientific › peer-review

22 Citations (Scopus)

Abstract

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

METHODS: Through RPTPα knockdown, we assessed FLS gene expression by quantitative polymerase chain reaction and enzyme-linked immunosorbent assay, invasion and migration in 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 Ptpra(-/-) 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 Y527 and decreased phosphorylation of FAK on stimulatory Y397. Treatment of RA FLS with an inhibitor of FAK phenocopied knockdown of RPTPα. Ptpra-deficient mice were protected from arthritis development, which was due to radioresistant cells.

CONCLUSIONS: By regulating phosphorylation of SRC and FAK, RPTPα mediates pro-inflammatory and pro-invasive signaling in RA FLS, correlating with promotion of disease in an FLS-dependent model of RA. This article is protected by copyright. All rights reserved.

Original language	English
Journal	Arthritis and Rheumatology
Early online date	28 Sept 2015
DOIs	https://doi.org/10.1002/art.39442
Publication status	Published - 25 Jan 2016

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10.1002/art.39442

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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., Aleman Muench, G. R., 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 alpha enhances rheumatoid synovial fibroblast signaling and promotes arthritis in mice. Arthritis and Rheumatology. https://doi.org/10.1002/art.39442

@article{214be86e49bb4a9fa215b0ebdfbcada2,

title = "Receptor protein tyrosine phosphatase alpha enhances rheumatoid synovial fibroblast signaling and promotes arthritis in mice",

abstract = "OBJECTIVE: During rheumatoid arthritis (RA), fibroblast-like synoviocytes (FLS) critically promote disease pathogenesis by aggressively invading the joint extracellular matrix. The focal adhesion kinase (FAK) signaling pathway is emerging as a contributor to RA FLS anomalous behavior. The receptor protein tyrosine phosphatase α (RPTPα), encoded by the PTPRA gene, is a key promoter of FAK signaling. Here we investigated whether RPTPα mediates FLS aggressiveness and RA pathogenesis.METHODS: Through RPTPα knockdown, we assessed FLS gene expression by quantitative polymerase chain reaction and enzyme-linked immunosorbent assay, invasion and migration in 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 Ptpra(-/-) 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 Y527 and decreased phosphorylation of FAK on stimulatory Y397. Treatment of RA FLS with an inhibitor of FAK phenocopied knockdown of RPTPα. Ptpra-deficient mice were protected from arthritis development, which was due to radioresistant cells.CONCLUSIONS: By regulating phosphorylation of SRC and FAK, RPTPα mediates pro-inflammatory and pro-invasive signaling in RA FLS, correlating with promotion of disease in an FLS-dependent model of RA. This article is protected by copyright. All rights reserved.",

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 {Aleman Muench}, {German R} 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} 2015, American College of Rheumatology.",

year = "2016",

month = jan,

day = "25",

doi = "10.1002/art.39442",

language = "English",

journal = "Arthritis and Rheumatology",

issn = "2326-5191",

publisher = "John Wiley and Sons Ltd",

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Stanford, SM, Svensson, MND, Sacchetti, C, Pilo, CA, Wu, DJ, Kiosses, WB, Hellvard, A, Bergum, B, Aleman Muench, GR, 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 alpha enhances rheumatoid synovial fibroblast signaling and promotes arthritis in mice', Arthritis and Rheumatology. https://doi.org/10.1002/art.39442

TY - JOUR

T1 - Receptor protein tyrosine phosphatase alpha enhances rheumatoid synovial fibroblast signaling and promotes 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 - Aleman Muench, German R

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 - 2016/1/25

Y1 - 2016/1/25

N2 - OBJECTIVE: During rheumatoid arthritis (RA), fibroblast-like synoviocytes (FLS) critically promote disease pathogenesis by aggressively invading the joint extracellular matrix. The focal adhesion kinase (FAK) signaling pathway is emerging as a contributor to RA FLS anomalous behavior. The receptor protein tyrosine phosphatase α (RPTPα), encoded by the PTPRA gene, is a key promoter of FAK signaling. Here we investigated whether RPTPα mediates FLS aggressiveness and RA pathogenesis.METHODS: Through RPTPα knockdown, we assessed FLS gene expression by quantitative polymerase chain reaction and enzyme-linked immunosorbent assay, invasion and migration in 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 Ptpra(-/-) 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 Y527 and decreased phosphorylation of FAK on stimulatory Y397. Treatment of RA FLS with an inhibitor of FAK phenocopied knockdown of RPTPα. Ptpra-deficient mice were protected from arthritis development, which was due to radioresistant cells.CONCLUSIONS: By regulating phosphorylation of SRC and FAK, RPTPα mediates pro-inflammatory and pro-invasive signaling in RA FLS, correlating with promotion of disease in an FLS-dependent model of RA. This article is protected by copyright. All rights reserved.

AB - OBJECTIVE: During rheumatoid arthritis (RA), fibroblast-like synoviocytes (FLS) critically promote disease pathogenesis by aggressively invading the joint extracellular matrix. The focal adhesion kinase (FAK) signaling pathway is emerging as a contributor to RA FLS anomalous behavior. The receptor protein tyrosine phosphatase α (RPTPα), encoded by the PTPRA gene, is a key promoter of FAK signaling. Here we investigated whether RPTPα mediates FLS aggressiveness and RA pathogenesis.METHODS: Through RPTPα knockdown, we assessed FLS gene expression by quantitative polymerase chain reaction and enzyme-linked immunosorbent assay, invasion and migration in 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 Ptpra(-/-) 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 Y527 and decreased phosphorylation of FAK on stimulatory Y397. Treatment of RA FLS with an inhibitor of FAK phenocopied knockdown of RPTPα. Ptpra-deficient mice were protected from arthritis development, which was due to radioresistant cells.CONCLUSIONS: By regulating phosphorylation of SRC and FAK, RPTPα mediates pro-inflammatory and pro-invasive signaling in RA FLS, correlating with promotion of disease in an FLS-dependent model of RA. This article is protected by copyright. All rights reserved.

U2 - 10.1002/art.39442

DO - 10.1002/art.39442

M3 - Article

C2 - 26414708

SN - 2326-5191

JO - Arthritis and Rheumatology

JF - Arthritis and Rheumatology

ER -

Receptor protein tyrosine phosphatase alpha enhances rheumatoid synovial fibroblast signaling and promotes arthritis in mice

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