Rheumatoid arthritis (RA) is an autoimmune synovitis that affects 0.5% of the world population, yet the key autoantigen targets remain unknown. Production of autoantibodies, such as the anti-citrullinated protein antibodies (ACPAs), is a hallmark of RA. However, which antigens the critical ACPAs and other RA- associated autoantibodies target remains largely unknown. Also unknown is how these ACPAs develop, how similar the ACPA repertoires are between different individuals with RA, and whether and how they contribute to the pathogenesis of RA. So far, the research field has lacked the means to comprehensively characterize the autoantibodies associated with a given disease and to then rationally winnow them to those that are important-that is, those that either drive the disease or serve as identifiers of the key antigens that trigger the pathogenic T-cell response. We have now developed "antibody repertoire capture" technology, a high- throughput method that allows us to do just that. Harnessing the power of next-generation sequencing, we have developed a novel method for barcoding all the cDNAs generated from individual antibody-expressing cells, thereby enabling high-throughput analysis of the paired heavy- and light-chain immunoglobulin genes expressed by single B cells, plasmablasts, or plasma cells. We hypothesize that we can elucidate the pathogenic autoantibody responses associated with RA by defining the antibody repertoire of plasmablasts and antigen-sorted memory B cells in the blood, and of plasmablasts and plasma cells in the synovium, of individuals with RA. We will then bioinformatically analyze the antibody sequences we obtain to generate evolutionary trees of the antibody repertoires and thereby identify and clone the affinity-matured antibodies that are likely the key autoantibodies. We will identify the antigens targeted by these recombinant, affinity-matured autoantibodies, investigate how their specific sequences develop, dissect their binding and immunostimulatory properties, and assess their pathogenicity. Success of this proposal would shed light on the development of the autoantibody response and identify the key autoantigens targeted in RA, findings that could lead to the development of new diagnostics and therapies for RA.

Public Health Relevance

Rheumatoid arthritis (RA) affects 0.5%, of the U.S. population. The autoantigen targets are unknown for RA and many other autoimmune diseases. Success of the proposed studies would provide insights into pathogenesis of, and could lead to development of new diagnostic and therapeutic approaches for, RA and other autoimmune diseases.

Agency
National Institute of Health (NIH)
Institute
National Institute of Arthritis and Musculoskeletal and Skin Diseases (NIAMS)
Type
Research Project (R01)
Project #
1R01AR063676-01A1
Application #
8579836
Study Section
Arthritis, Connective Tissue and Skin Study Section (ACTS)
Program Officer
Mao, Su-Yau
Project Start
2013-09-01
Project End
2018-08-31
Budget Start
2013-09-01
Budget End
2014-08-31
Support Year
1
Fiscal Year
2013
Total Cost
$334,365
Indirect Cost
$121,865
Name
Stanford University
Department
Internal Medicine/Medicine
Type
Schools of Medicine
DUNS #
009214214
City
Stanford
State
CA
Country
United States
Zip Code
94305
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Kim, Yong-Gil; Sohn, Dong Hyun; Zhao, Xiaoyan et al. (2014) Role of protein phosphatase magnesium-dependent 1A and anti-protein phosphatase magnesium-dependent 1A autoantibodies in ankylosing spondylitis. Arthritis Rheumatol 66:2793-803
Tan, Yann-Chong; Blum, Lisa K; Kongpachith, Sarah et al. (2014) High-throughput sequencing of natively paired antibody chains provides evidence for original antigenic sin shaping the antibody response to influenza vaccination. Clin Immunol 151:55-65
Sokolove, Jeremy; Johnson, Dannette S; Lahey, Lauren J et al. (2014) Rheumatoid factor as a potentiator of anti-citrullinated protein antibody-mediated inflammation in rheumatoid arthritis. Arthritis Rheumatol 66:813-21
Lu, Daniel R; Tan, Yann-Chong; Kongpachith, Sarah et al. (2014) Identifying functional anti-Staphylococcus aureus antibodies by sequencing antibody repertoires of patient plasmablasts. Clin Immunol 152:77-89