ACR Meeting Abstracts

ACR Meeting Abstracts

Abstract Number: 0460

Single-cell Profiling of B and T Cell Repertoire and Gene Expression in the RA Synovium Reveals Tissue Specific Clonal Expansion

Nida Meednu1, Aaron Wagner2, Garett Dunlap3, Fan Zhang4, Anna Helena Jonsson5, Kevin Wei5, Paul Utz6, William Robinson7, Holden Maecker7, Judith James8, Joel Guthridge8, S. Louis Bridges, Jr.9, Vivian Bykerk9, Laura Donlin9, Susan Goodman9, Edward DiCarlo9, Christopher Ritchlin10, Darren Tabechian2, James Lederer11, Ellen Gravallese12, Mandy McGeachy13, Gary Firestein14, David Boyle15, Peter Gregersen16, Diane Horowitz17, Harris Perlman18, Arthur Mandelin18, Joan Bathon19, Laura Geraldino-Pardilla19, Laura Hughes20, V. Michael Holers21, Kevin Deane22, Larry Moreland21, Andrew Filer23, Costantino Pitzalis24, Lindsy Forbess25, Ami Ben-artzi26, Karen Salomon-Escoto27, Soumya Raychaudhuri5, Michael Brenner28, Deepak Rao5, Andrew McDavid2, Jennifer Anolik1 and Accelerating Medicines Partership (AMP) RA/SLE Network29, 1University of Rochester Medical center, Rochester, NY, 2University of Rochester, Rochester, NY, 3Harvard University, Somerville, MA, 4Harvard Medical School, Boston, MA, 5Brigham and Women's Hospital, Boston, MA, 6Stanford University, Stanford, CA, 7Stanford University, Palo Alto, CA, 8Oklahoma Medical Research Foundation, Oklahoma City, OK, 9Hospital for Special Surgery, New York, NY, 10Division of Allergy, Immunology, and Rheumatology, School of Medicine and Dentistry, University of Rochester Medical Center, Rochester, NY, 11Brigham and Women's Hospital and Harvard Medical School, Millis, MA, 12Brigham and Women's Hospital, Harvard Medical School, Chestnut Hill, MA, 13University of Pittsburgh, Pittsburgh, PA, 14University of California San Diego, San Diego, CA, 15University of California San Diego, La Jolla, CA, 16The Feinstein Institute for Medical Research, Larchmont, NY, 17Northwell Health, Jericho, NY, 18Northwestern University, Chicago, IL, 19Columbia University, New York, NY, 20University of Alabama at Birmingham, Birmingham, AL, 21University of Colorado, Denver, CO, 22University of Colorado Denver, Denver, CO, 23University of Birmingham, Birmingham, United Kingdom, 24Queen Mary University of London, London, United Kingdom, 25Cedars-Sinai Medical Center, Los Angeles, CA, 26Cedars-Sinai Medical Center, Beverly Hills, CA, 27University of Massachusetts Medical School, Shrewsbury, MA, 28Brigham and Women's Hospital, Harvard Medical School, Newton, MA, 29Brigham and Women's Hospital, Everett, MA

Meeting: ACR Convergence 2021

Keywords: , , , ,

Session Information

Date: Saturday, November 6, 2021

Title: Abstracts: RA – Etiology & Pathogenesis (0458–0461)

Session Type: Abstract Session

Session Time: 11:00AM-11:15AM

Background/Purpose: B cell and T cell activation pathways in the synovium are an incompletely understood feature of rheumatoid arthritis. In this study, utilizing single cell RNA sequencing coupled with B and T cell repertoire sequencing to characterize B and T cell states that may play a role in immune activation within the synovium, we define potential developmental relationships and identify key signaling pathways that support local immune cell activation and development of autoreactive plasma cells.

Methods: B and T cells were sorted by flow cytometry from RA synovial tissue (n=13) and matched peripheral blood (PBL) (n=10) as part of the AMP Network Phase 2. Single cell RNA sequencing was performed on the 10x Genomics platform with poly-A selected, 5’ initiated expression and BCR or TCR libraries generated from each single cell. Quality control criteria (# of genes detected and % mitochondria gene) were applied to the sequencing data. Clustering analysis using gene expression was performed to identify B cell and T cell subsets. In B cells, we analyzed somatic hypermutation (SHM), immunoglobulin isotype and clonality. Diversity was assessed by looking at V(D)J segment usage frequencies and CDR3 properties. In T cells, we assessed the extent of oligoclonality in different T cell subsets and the overlap in repertoires across T cell subsets.

Results: After applying QC, over 50,000 lymphocytes were available for downstream analysis (36,000 from PBL and 20,600 cells from synovial tissue). Clustering identified various naive and activated B cell populations, plasma cells, and naive CD4 and CD8, Treg and effector T cell states. We observed synovial enrichment of clusters including activated B cells, Tph cells, and granzyme K-expressing CD8 T cells. Activated synovial B cells had significantly higher SHM compared to PBL. In every subject, we observed substantial clonal expansion in both the synovium and peripheral blood, as well as clones shared between these tissues. Naïve B and T cells exhibited fewer expanded clonotypes than non-naïve populations. In both B cells and T cells, there was clonal sharing across synovial sub-populations, including between IgG and IgA plasma cells, activated B cell and plasma cells, and between memory and effector T cells. Together, our findings suggest in situ differentiation of selected B and T cell clones and trafficking of these expanded clones between blood and synovium.

Conclusion: Our data demonstrate the value of integrating gene expression and repertoire data for the study of T and B cell responses in RA synovial tissue and provide evidence of in situ selection.

Disclosures: N. Meednu, None; A. Wagner, None; G. Dunlap, None; F. Zhang, None; A. Jonsson, Amgen, 5; K. Wei, Mestag, 2; P. Utz, None; W. Robinson, None; H. Maecker, None; J. James, Progentec Diagnostics, Inc., 2; J. Guthridge, None; S. Bridges, Jr., None; V. Bykerk, Amgen Inc., 2, 6, Bristol Myers Squibb, 2, 6, Gilead, 2, 6, Pfizer, 2, 6, Regeneron, 2, 6, Sanofi-Genzyme, 2, 6, UCB, 2, 6; L. Donlin, Karius, Inc., 5, Stryker, 2, 6; S. Goodman, UCB, 1, NOvartis, 5; E. DiCarlo, None; C. Ritchlin, UCB, 2, 5, AbbVie, 2, 5, Amgen, 2, 5, Eli Lilly, 2, Pfizer, 2, Novartis, 2, Gilead, 2, Janssen, 2; D. Tabechian, Amgen, 11; J. Lederer, None; E. Gravallese, None; M. McGeachy, None; G. Firestein, Eli Lilly, 5; D. Boyle, Janssen R&D, 5; P. Gregersen, None; D. Horowitz, None; H. Perlman, Kiniksa, 1, 2; A. Mandelin, AbbVie, 6, CVS CareMark, 2, Horizon, 6, Lilly, 6, Pfizer, 6, Sanofi Genzyme / Regeneron, 6, UCB, 6; J. Bathon, None; L. Geraldino-Pardilla, None; L. Hughes, None; V. Holers, Jansson, 5; K. Deane, Inova Diagnostics, Inc, 5, Bristol Meyers Squibb, 1, 5, Janssen Research and Development, LLC, 5, imaware, 2, ThermoFisher, 2, 5, Medscape, 6; L. Moreland, None; A. Filer, GSK, 5, Mestag, 5, Galapagos, 2, 5, Janssen, 5, Roche, 5, Nascient, 5; C. Pitzalis, None; L. Forbess, None; A. Ben-artzi, None; K. Salomon-Escoto, None; S. Raychaudhuri, Mestag Therapeutics, 2, 12, Founder, Johnson & Johnson, 1, 2, Pfizer, 1, 2, Biogen, 5, Gilead Sciences, 2; M. Brenner, GSK, 2, 4FO Ventures, 2, Mestag Therapeutics, 2, 11; D. Rao, Janssen, 5, 6, Bristol-Myers Squibb, 1, 5, Scipher Medicine, 2, Pfizer, 6, Merck, 6; A. McDavid, None; J. Anolik, None; A. (AMP) RA/SLE Network, None.

To cite this abstract in AMA style:

Meednu N, Wagner A, Dunlap G, Zhang F, Jonsson A, Wei K, Utz P, Robinson W, Maecker H, James J, Guthridge J, Bridges, Jr. S, Bykerk V, Donlin L, Goodman S, DiCarlo E, Ritchlin C, Tabechian D, Lederer J, Gravallese E, McGeachy M, Firestein G, Boyle D, Gregersen P, Horowitz D, Perlman H, Mandelin A, Bathon J, Geraldino-Pardilla L, Hughes L, Holers V, Deane K, Moreland L, Filer A, Pitzalis C, Forbess L, Ben-artzi A, Salomon-Escoto K, Raychaudhuri S, Brenner M, Rao D, McDavid A, Anolik J, (AMP) RA/SLE Network A. Single-cell Profiling of B and T Cell Repertoire and Gene Expression in the RA Synovium Reveals Tissue Specific Clonal Expansion [abstract]. Arthritis Rheumatol. 2021; 73 (suppl 9). https://acrabstracts.org/abstract/single-cell-profiling-of-b-and-t-cell-repertoire-and-gene-expression-in-the-ra-synovium-reveals-tissue-specific-clonal-expansion/. Accessed .

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