Proteomic analysis of human osteoarthritis synovial fluid

Lavanya Balakrishnan; Raja Sekhar Nirujogi; Sartaj Ahmad; Mitali Bhattacharjee; Srikanth S. Manda; Santosh Renuse; Dhanashree S. Kelkar; Yashwanth Subbannayya; Rajesh Raju; Renu Goel; Joji Kurian Thomas; Navjyot Kaur; Mukesh Dhillon; Shantal Gupta Tankala; Ramesh Jois; Vivek Vasdev; Y. L. Ramachandra; Nandini A. Sahasrabuddhe; T. S.Keshava Prasad; Sujatha Mohan; Harsha Gowda; Subramanian Shankar; Akhilesh Pandey

doi:10.1186/1559-0275-11-6

Proteomic analysis of human osteoarthritis synovial fluid

Lavanya Balakrishnan, Raja Sekhar Nirujogi, Sartaj Ahmad, Mitali Bhattacharjee, Srikanth S. Manda, Santosh Renuse, Dhanashree S. Kelkar, Yashwanth Subbannayya, Rajesh Raju, Renu Goel, Joji Kurian Thomas, Navjyot Kaur, Mukesh Dhillon, Shantal Gupta Tankala, Ramesh Jois, Vivek Vasdev, Y. L. Ramachandra, Nandini A. Sahasrabuddhe, T. S.Keshava Prasad, Sujatha MohanHarsha Gowda, Subramanian Shankar^*, Akhilesh Pandey

^*Corresponding author for this work

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

112 Scopus citations

Abstract

Background: Osteoarthritis is a chronic musculoskeletal disorder characterized mainly by progressive degradation of the hyaline cartilage. Patients with osteoarthritis often postpone seeking medical help, which results in the diagnosis being made at an advanced stage of cartilage destruction. Sustained efforts are needed to identify specific markers that might help in early diagnosis, monitoring disease progression and in improving therapeutic outcomes. We employed a multipronged proteomic approach, which included multiple fractionation strategies followed by high resolution mass spectrometry analysis to explore the proteome of synovial fluid obtained from osteoarthritis patients. In addition to the total proteome, we also enriched glycoproteins from synovial fluid using lectin affinity chromatography. Results: We identified 677 proteins from synovial fluid of patients with osteoarthritis of which 545 proteins have not been previously reported. These novel proteins included ADAM-like decysin 1 (ADAMDEC1), alanyl (membrane) aminopeptidase (ANPEP), CD84, fibulin 1 (FBLN1), matrix remodelling associated 5 (MXRA5), secreted phosphoprotein 2 (SPP2) and spondin 2 (SPON2). We identified 300 proteins using lectin affinity chromatography, including the glycoproteins afamin (AFM), attractin (ATRN), fibrillin 1 (FBN1), transferrin (TF), tissue inhibitor of metalloproteinase 1 (TIMP1) and vasorin (VSN). Gene ontology analysis confirmed that a majority of the identified proteins were extracellular and are mostly involved in cell communication and signaling. We also confirmed the expression of ANPEP, dickkopf WNT signaling pathway inhibitor 3 (DKK3) and osteoglycin (OGN) by multiple reaction monitoring (MRM) analysis of osteoarthritis synovial fluid samples. Conclusions: We present an in-depth analysis of the synovial fluid proteome from patients with osteoarthritis. We believe that the catalog of proteins generated in this study will further enhance our knowledge regarding the pathophysiology of osteoarthritis and should assist in identifying better biomarkers for early diagnosis.

Original language	English (US)
Article number	6
Journal	Clinical Proteomics
Volume	11
Issue number	1
DOIs	https://doi.org/10.1186/1559-0275-11-6
State	Published - 2014
Externally published	Yes

Funding

We thank the Department of Biotechnology, Government of India for research support to the Institute of Bioinformatics, Bangalore. We thank Agilent Technologies and Thermo Scientific for the instrument support. Raja Sekhar Nirujogi is a recipient of Senior Research Fellowship award from Council of Scientific and Industrial Research (CSIR), Government of India. Sartaj Ahmad is a recipient of Junior Research Fellowship from University Grants Commission (UGC), Government of India. Srikanth Srinivas Manda. Santosh Renuse and Dhanashree S. Kelkar are recipients of Senior Research Fellowship award from the University Grants Commission (UGC), Government of India. Harsha Gowda is a Wellcome Trust/DBT India Alliance Early Career Fellow.

Keywords

Body fluid
Cartilage
Glycosylation
Joint destruction

ASJC Scopus subject areas

Molecular Medicine
Molecular Biology
Clinical Biochemistry

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10.1186/1559-0275-11-6

Cite this

Balakrishnan, L., Nirujogi, R. S., Ahmad, S., Bhattacharjee, M., Manda, S. S., Renuse, S., Kelkar, D. S., Subbannayya, Y., Raju, R., Goel, R., Thomas, J. K., Kaur, N., Dhillon, M., Tankala, S. G., Jois, R., Vasdev, V., Ramachandra, Y. L., Sahasrabuddhe, N. A., Prasad, T. S. K., ... Pandey, A. (2014). Proteomic analysis of human osteoarthritis synovial fluid. Clinical Proteomics, 11(1), Article 6. https://doi.org/10.1186/1559-0275-11-6

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title = "Proteomic analysis of human osteoarthritis synovial fluid",

abstract = "Background: Osteoarthritis is a chronic musculoskeletal disorder characterized mainly by progressive degradation of the hyaline cartilage. Patients with osteoarthritis often postpone seeking medical help, which results in the diagnosis being made at an advanced stage of cartilage destruction. Sustained efforts are needed to identify specific markers that might help in early diagnosis, monitoring disease progression and in improving therapeutic outcomes. We employed a multipronged proteomic approach, which included multiple fractionation strategies followed by high resolution mass spectrometry analysis to explore the proteome of synovial fluid obtained from osteoarthritis patients. In addition to the total proteome, we also enriched glycoproteins from synovial fluid using lectin affinity chromatography. Results: We identified 677 proteins from synovial fluid of patients with osteoarthritis of which 545 proteins have not been previously reported. These novel proteins included ADAM-like decysin 1 (ADAMDEC1), alanyl (membrane) aminopeptidase (ANPEP), CD84, fibulin 1 (FBLN1), matrix remodelling associated 5 (MXRA5), secreted phosphoprotein 2 (SPP2) and spondin 2 (SPON2). We identified 300 proteins using lectin affinity chromatography, including the glycoproteins afamin (AFM), attractin (ATRN), fibrillin 1 (FBN1), transferrin (TF), tissue inhibitor of metalloproteinase 1 (TIMP1) and vasorin (VSN). Gene ontology analysis confirmed that a majority of the identified proteins were extracellular and are mostly involved in cell communication and signaling. We also confirmed the expression of ANPEP, dickkopf WNT signaling pathway inhibitor 3 (DKK3) and osteoglycin (OGN) by multiple reaction monitoring (MRM) analysis of osteoarthritis synovial fluid samples. Conclusions: We present an in-depth analysis of the synovial fluid proteome from patients with osteoarthritis. We believe that the catalog of proteins generated in this study will further enhance our knowledge regarding the pathophysiology of osteoarthritis and should assist in identifying better biomarkers for early diagnosis.",

keywords = "Body fluid, Cartilage, Glycosylation, Joint destruction",

author = "Lavanya Balakrishnan and Nirujogi, {Raja Sekhar} and Sartaj Ahmad and Mitali Bhattacharjee and Manda, {Srikanth S.} and Santosh Renuse and Kelkar, {Dhanashree S.} and Yashwanth Subbannayya and Rajesh Raju and Renu Goel and Thomas, {Joji Kurian} and Navjyot Kaur and Mukesh Dhillon and Tankala, {Shantal Gupta} and Ramesh Jois and Vivek Vasdev and Ramachandra, {Y. L.} and Sahasrabuddhe, {Nandini A.} and Prasad, {T. S.Keshava} and Sujatha Mohan and Harsha Gowda and Subramanian Shankar and Akhilesh Pandey",

note = "Funding Information: We thank the Department of Biotechnology, Government of India for research support to the Institute of Bioinformatics, Bangalore. We thank Agilent Technologies and Thermo Scientific for the instrument support. Raja Sekhar Nirujogi is a recipient of Senior Research Fellowship award from Council of Scientific and Industrial Research (CSIR), Government of India. Sartaj Ahmad is a recipient of Junior Research Fellowship from University Grants Commission (UGC), Government of India. Srikanth Srinivas Manda. Santosh Renuse and Dhanashree S. Kelkar are recipients of Senior Research Fellowship award from the University Grants Commission (UGC), Government of India. Harsha Gowda is a Wellcome Trust/DBT India Alliance Early Career Fellow.",

year = "2014",

doi = "10.1186/1559-0275-11-6",

language = "English (US)",

volume = "11",

journal = "Clinical Proteomics",

issn = "1542-6416",

publisher = "BioMed Central",

number = "1",

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Balakrishnan, L, Nirujogi, RS, Ahmad, S, Bhattacharjee, M, Manda, SS, Renuse, S, Kelkar, DS, Subbannayya, Y, Raju, R, Goel, R, Thomas, JK, Kaur, N, Dhillon, M, Tankala, SG, Jois, R, Vasdev, V, Ramachandra, YL, Sahasrabuddhe, NA, Prasad, TSK, Mohan, S, Gowda, H, Shankar, S & Pandey, A 2014, 'Proteomic analysis of human osteoarthritis synovial fluid', Clinical Proteomics, vol. 11, no. 1, 6. https://doi.org/10.1186/1559-0275-11-6

TY - JOUR

T1 - Proteomic analysis of human osteoarthritis synovial fluid

AU - Balakrishnan, Lavanya

AU - Nirujogi, Raja Sekhar

AU - Ahmad, Sartaj

AU - Bhattacharjee, Mitali

AU - Manda, Srikanth S.

AU - Renuse, Santosh

AU - Kelkar, Dhanashree S.

AU - Subbannayya, Yashwanth

AU - Raju, Rajesh

AU - Goel, Renu

AU - Thomas, Joji Kurian

AU - Kaur, Navjyot

AU - Dhillon, Mukesh

AU - Tankala, Shantal Gupta

AU - Jois, Ramesh

AU - Vasdev, Vivek

AU - Ramachandra, Y. L.

AU - Sahasrabuddhe, Nandini A.

AU - Prasad, T. S.Keshava

AU - Mohan, Sujatha

AU - Gowda, Harsha

AU - Shankar, Subramanian

AU - Pandey, Akhilesh

N1 - Funding Information: We thank the Department of Biotechnology, Government of India for research support to the Institute of Bioinformatics, Bangalore. We thank Agilent Technologies and Thermo Scientific for the instrument support. Raja Sekhar Nirujogi is a recipient of Senior Research Fellowship award from Council of Scientific and Industrial Research (CSIR), Government of India. Sartaj Ahmad is a recipient of Junior Research Fellowship from University Grants Commission (UGC), Government of India. Srikanth Srinivas Manda. Santosh Renuse and Dhanashree S. Kelkar are recipients of Senior Research Fellowship award from the University Grants Commission (UGC), Government of India. Harsha Gowda is a Wellcome Trust/DBT India Alliance Early Career Fellow.

PY - 2014

Y1 - 2014

N2 - Background: Osteoarthritis is a chronic musculoskeletal disorder characterized mainly by progressive degradation of the hyaline cartilage. Patients with osteoarthritis often postpone seeking medical help, which results in the diagnosis being made at an advanced stage of cartilage destruction. Sustained efforts are needed to identify specific markers that might help in early diagnosis, monitoring disease progression and in improving therapeutic outcomes. We employed a multipronged proteomic approach, which included multiple fractionation strategies followed by high resolution mass spectrometry analysis to explore the proteome of synovial fluid obtained from osteoarthritis patients. In addition to the total proteome, we also enriched glycoproteins from synovial fluid using lectin affinity chromatography. Results: We identified 677 proteins from synovial fluid of patients with osteoarthritis of which 545 proteins have not been previously reported. These novel proteins included ADAM-like decysin 1 (ADAMDEC1), alanyl (membrane) aminopeptidase (ANPEP), CD84, fibulin 1 (FBLN1), matrix remodelling associated 5 (MXRA5), secreted phosphoprotein 2 (SPP2) and spondin 2 (SPON2). We identified 300 proteins using lectin affinity chromatography, including the glycoproteins afamin (AFM), attractin (ATRN), fibrillin 1 (FBN1), transferrin (TF), tissue inhibitor of metalloproteinase 1 (TIMP1) and vasorin (VSN). Gene ontology analysis confirmed that a majority of the identified proteins were extracellular and are mostly involved in cell communication and signaling. We also confirmed the expression of ANPEP, dickkopf WNT signaling pathway inhibitor 3 (DKK3) and osteoglycin (OGN) by multiple reaction monitoring (MRM) analysis of osteoarthritis synovial fluid samples. Conclusions: We present an in-depth analysis of the synovial fluid proteome from patients with osteoarthritis. We believe that the catalog of proteins generated in this study will further enhance our knowledge regarding the pathophysiology of osteoarthritis and should assist in identifying better biomarkers for early diagnosis.

AB - Background: Osteoarthritis is a chronic musculoskeletal disorder characterized mainly by progressive degradation of the hyaline cartilage. Patients with osteoarthritis often postpone seeking medical help, which results in the diagnosis being made at an advanced stage of cartilage destruction. Sustained efforts are needed to identify specific markers that might help in early diagnosis, monitoring disease progression and in improving therapeutic outcomes. We employed a multipronged proteomic approach, which included multiple fractionation strategies followed by high resolution mass spectrometry analysis to explore the proteome of synovial fluid obtained from osteoarthritis patients. In addition to the total proteome, we also enriched glycoproteins from synovial fluid using lectin affinity chromatography. Results: We identified 677 proteins from synovial fluid of patients with osteoarthritis of which 545 proteins have not been previously reported. These novel proteins included ADAM-like decysin 1 (ADAMDEC1), alanyl (membrane) aminopeptidase (ANPEP), CD84, fibulin 1 (FBLN1), matrix remodelling associated 5 (MXRA5), secreted phosphoprotein 2 (SPP2) and spondin 2 (SPON2). We identified 300 proteins using lectin affinity chromatography, including the glycoproteins afamin (AFM), attractin (ATRN), fibrillin 1 (FBN1), transferrin (TF), tissue inhibitor of metalloproteinase 1 (TIMP1) and vasorin (VSN). Gene ontology analysis confirmed that a majority of the identified proteins were extracellular and are mostly involved in cell communication and signaling. We also confirmed the expression of ANPEP, dickkopf WNT signaling pathway inhibitor 3 (DKK3) and osteoglycin (OGN) by multiple reaction monitoring (MRM) analysis of osteoarthritis synovial fluid samples. Conclusions: We present an in-depth analysis of the synovial fluid proteome from patients with osteoarthritis. We believe that the catalog of proteins generated in this study will further enhance our knowledge regarding the pathophysiology of osteoarthritis and should assist in identifying better biomarkers for early diagnosis.

KW - Body fluid

KW - Cartilage

KW - Glycosylation

KW - Joint destruction

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JO - Clinical Proteomics

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Proteomic analysis of human osteoarthritis synovial fluid

Abstract

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Keywords

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