Publications by authors named "Peter Gregersen"

286 Publications

Arg206Cys substitution in causes a defect in DNASE1L3 protein secretion that confers risk of systemic lupus erythematosus.

Authors:
Latanya N Coke Hongxiu Wen Mary Comeau Mustafa H Ghanem Andrew Shih Christine N Metz Wentian Li Carl D Langefeld Peter K Gregersen Kim R Simpfendorfer

Ann Rheum Dis 2021 Jan 17. Epub 2021 Jan 17.

The Institute of Molecular Medicine, Northwell Health Feinstein Institutes for Medical Research, Manhasset, New York, USA

Objectives: To determine if the polymorphism encoding the Arg206Cys substitution in DNASE1L3 explains the association of the / gene locus with systemic lupus erythematosus (SLE) and to examine the effect of the Arg206Cys sequence change on DNASE1L3 protein function.

Methods: Conditional analysis for rs35677470 was performed on cases and controls with European ancestry from the SLE Immunochip study, and genotype and haplotype frequencies were compared. DNASE1L3 protein levels were measured in cells and supernatants of HEK293 cells and monocyte-derived dendritic cells expressing recombinant and endogenous 206Arg and 206Cys protein variants.

Results: Conditional analysis on rs35677470 eliminated the SLE risk association signal for lead single-nucleotide polymorphisms (SNPs) rs180977001 and rs73081554, which are found to tag the same risk haplotype as rs35677470. The modest effect sizes of the SLE risk genotypes (heterozygous risk OR=1.14 and homozygous risk allele OR=1.68) suggest some DNASE1L3 endonuclease enzyme function is retained. An SLE protective signal in (lead SNP rs11130643) remained following conditioning on rs35677470. The DNASE1L3 206Cys risk variant maintained enzymatic activity, but secretion of the artificial and endogenous DNASE1L3 206Cys protein was substantially reduced.

Conclusions: SLE risk association in the locus is dependent on the missense SNP rs35677470, which confers a reduction in DNASE1L3 protein secretion but does not eliminate its DNase enzyme function.
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http://dx.doi.org/10.1136/annrheumdis-2020-218810DOI Listing
January 2021

Diminished cytokine-induced Jak/STAT signaling is associated with rheumatoid arthritis and disease activity.

Authors:
Jason Ptacek Rachael E Hawtin Dongmei Sun Brent Louie Erik Evensen Barbara B Mittleman Alessandra Cesano Guy Cavet Clifton O Bingham Stacey S Cofield Jeffrey R Curtis Maria I Danila Chander Raman Richard A Furie Mark C Genovese William H Robinson Marc C Levesque Larry W Moreland Peter A Nigrovic Nancy A Shadick James R O'Dell Geoffrey M Thiele E William St Clair Christopher C Striebich Matthew B Hale Houman Khalili Franak Batliwalla Cynthia Aranow Meggan Mackay Betty Diamond Garry P Nolan Peter K Gregersen S Louis Bridges

PLoS One 2021 14;16(1):e0244187. Epub 2021 Jan 14.

Hospital for Special Surgery and Weill Cornell Medical College, New York, New York, United States of America.

Rheumatoid arthritis (RA) is a systemic and incurable autoimmune disease characterized by chronic inflammation in synovial lining of joints. To identify the signaling pathways involved in RA, its disease activity, and treatment response, we adapted a systems immunology approach to simultaneously quantify 42 signaling nodes in 21 immune cell subsets (e.g., IFNα→p-STAT5 in B cells) in peripheral blood mononuclear cells (PBMC) from 194 patients with longstanding RA (including 98 patients before and after treatment), and 41 healthy controls (HC). We found multiple differences between patients with RA compared to HC, predominantly in cytokine-induced Jak/STAT signaling in many immune cell subsets, suggesting pathways that may be associated with susceptibility to RA. We also found that high RA disease activity, compared to low disease activity, was associated with decreased (e.g., IFNα→p-STAT5, IL-10→p-STAT1) or increased (e.g., IL-6→STAT3) response to stimuli in multiple cell subsets. Finally, we compared signaling in patients with established, refractory RA before and six months after initiation of methotrexate (MTX) or TNF inhibitors (TNFi). We noted significant changes from pre-treatment to post-treatment in IFNα→p-STAT5 signaling and IL-10→p-STAT1 signaling in multiple cell subsets; these changes brought the aberrant RA signaling profiles toward those of HC. This large, comprehensive functional signaling pathway study provides novel insights into the pathogenesis of RA and shows the potential of quantification of cytokine-induced signaling as a biomarker of disease activity or treatment response.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0244187PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7808603PMC
January 2021

Association of lipid mediators and development of future incident inflammatory arthritis in an anti-citrullinated protein antibody positive population.

Authors:
Kristen J Polinski Elizabeth A Bemis Fan Yang Tessa Crume M Kristen Demoruelle Marie Feser Jennifer Seifert James R O'Dell Ted R Mikuls Michael H Weisman Peter K Gregersen Richard M Keating Jane Buckner Nichole Reisdorph Kevin D Deane Michael Clare-Salzler V Michael Holers Jill M Norris

Arthritis Rheumatol 2020 Dec 31. Epub 2020 Dec 31.

Department of Epidemiology, Colorado School of Public Health, Aurora, CO, United States.

Objective: To determine the association of polyunsaturated fatty acid (PUFA)-derived lipid mediators with progression from rheumatoid arthritis (RA)-related autoimmunity to inflammatory arthritis (IA).

Methods: We conducted a prospective cohort study using data from the Studies of the Etiologies of Rheumatoid Arthritis (SERA). SERA enrolled first-degree relatives (FDRs) of individuals with RA (FDR cohort) and individuals who screened positive for RA-related autoantibodies at health fairs (screened cohort). We followed 135 anti-CCP3.1 positive participants, of which 29 developed IA. Lipid mediators selected a priori were quantified from stored plasma samples using liquid chromatography tandem mass spectrometry. We fit multivariable Cox proportional hazard models for each lipid mediator as a time-varying variable. For lipid mediators found to be significantly associated with IA, we then examined IL-1β, IL-6, IL-8 and TNF-α as potential statistical mediators.

Results: For every one natural log pg/ml increase in the circulating plasma levels of pro-inflammatory 5-HETE, the risk of developing IA increases by 241% (HR: 2.41; 95% CI: 1.43, 4.07) after adjusting for age at baseline, cohort (FDR or screened), and shared epitope status. The models examining 15-HETE and 17-HDHA had the same trend but did not reach significance. We did not find evidence that the association between 5-HETE and IA risk was influenced by the tested pro-inflammatory cytokines.

Conclusion: In a prospective cohort of anti-CCP positive individuals, higher levels of 5-HETE, an important precursor to pro-inflammatory leukotrienes, was associated with subsequent IA. Our findings highlight the potential significance of these PUFA metabolites in pre-RA populations.
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http://dx.doi.org/10.1002/art.41631DOI Listing
December 2020

Deep sequencing reveals a DAP1 regulatory haplotype that potentiates autoimmunity in systemic lupus erythematosus.

Authors:
Prithvi Raj Ran Song Honglin Zhu Linley Riediger Dong-Jae Jun Chaoying Liang Carlos Arana Bo Zhang Yajing Gao Benjamin E Wakeland Igor Dozmorov Jinchun Zhou Jennifer A Kelly Bernard R Lauwerys Joel M Guthridge Nancy J Olsen Swapan K Nath Chandrashekhar Pasare Nicolai van Oers Gary Gilkeson Betty P Tsao Patrick M Gaffney Peter K Gregersen Judith A James Xiaoxia Zuo David R Karp Quan-Zhen Li Edward K Wakeland

Genome Biol 2020 Nov 19;21(1):281. Epub 2020 Nov 19.

Department of Immunology, University of Texas Southwestern Medical Center, Dallas, TX, 75390, USA.

Background: Systemic lupus erythematosus (SLE) is a clinically heterogeneous autoimmune disease characterized by the development of anti-nuclear antibodies. Susceptibility to SLE is multifactorial, with a combination of genetic and environmental risk factors contributing to disease development. Like other polygenic diseases, a significant proportion of estimated SLE heritability is not accounted for by common disease alleles analyzed by SNP array-based GWASs. Death-associated protein 1 (DAP1) was implicated as a candidate gene in a previous familial linkage study of SLE and rheumatoid arthritis, but the association has not been explored further.

Results: We perform deep sequencing across the DAP1 genomic segment in 2032 SLE patients, and healthy controls, and discover a low-frequency functional haplotype strongly associated with SLE risk in multiple ethnicities. We find multiple cis-eQTLs embedded in a risk haplotype that progressively downregulates DAP1 transcription in immune cells. Decreased DAP1 transcription results in reduced DAP1 protein in peripheral blood mononuclear cells, monocytes, and lymphoblastoid cell lines, leading to enhanced autophagic flux in immune cells expressing the DAP1 risk haplotype. Patients with DAP1 risk allele exhibit significantly higher autoantibody titers and altered expression of the immune system, autophagy, and apoptosis pathway transcripts, indicating that the DAP1 risk allele mediates enhanced autophagy, leading to the survival of autoreactive lymphocytes and increased autoantibody.

Conclusions: We demonstrate how targeted sequencing captures low-frequency functional risk alleles that are missed by SNP array-based studies. SLE patients with the DAP1 genotype have distinct autoantibody and transcription profiles, supporting the dissection of SLE heterogeneity by genetic analysis.
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http://dx.doi.org/10.1186/s13059-020-02184-zDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7677828PMC
November 2020

Factors associated with progression to inflammatory arthritis in first-degree relatives of individuals with RA following autoantibody positive screening in a non-clinical setting.

Authors:
Elizabeth A Bemis M Kristen Demoruelle Jennifer A Seifert Kristen J Polinski Michael H Weisman Jane H Buckner Peter K Gregersen Ted R Mikuls James R ODell Richard M Keating Kevin D Deane V Michael Holers Jill M Norris

Ann Rheum Dis 2021 02 14;80(2):154-161. Epub 2020 Sep 14.

Department of Epidemiology, Colorado School of Public Health, Aurora, Colorado, USA

Objectives: Little is known about the likelihood of developing inflammatory arthritis (IA) in individuals who screen autoantibody positive (aAb+) in a non-clinical research setting.

Methods: We screened for serum cyclic citrullinated peptide antibody (anti-CCP) and rheumatoid factor isotype aAbs in subjects who were at increased risk for rheumatoid arthritis (RA) because they are a first-degree relative of an individual with classified RA (n=1780). We evaluated combinations of aAbs and high titre aAbs, as defined by 2-times (2 x) the standard cut-off and an optimal cut-off, as predictors of our two outcomes, aAb+ persistence and incident IA.

Results: 304 subjects (17.1%) tested aAb+; of those, 131 were IA-free and had at least one follow-up visit. Sixty-four per cent of these tested aAb+ again on their next visit. Anti-CCP+ at levels ≥2 x the standard cut-off was associated with 13-fold higher likelihood of aAb +persistence. During a median of 4.4 years (IQR: 2.2-7.2), 20 subjects (15.3%) developed IA. Among subjects that screened anti-CCP+ at ≥ 2 x or ≥an optimal cut-off, 32% and 26% had developed IA within 5 years, respectively. Both anti-CCP cut-offs conferred an approximate fourfold increased risk of future IA (HR 4.09 and HR 3.95, p<0.01).

Conclusions: These findings support that aAb screening in a non-clinical setting can identify RA-related aAb+ individuals, as well as levels and combinations of aAbs that are associated with higher risk for future IA. Monitoring for the development of IA in aAb+ individuals and similar aAb testing approaches in at-risk populations may identify candidates for prevention studies in RA.
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http://dx.doi.org/10.1136/annrheumdis-2020-217066DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7855648PMC
February 2021

Menstruation: science and society.

Authors:
Hilary O D Critchley Elnur Babayev Serdar E Bulun Sandy Clark Iolanda Garcia-Grau Peter K Gregersen Aoife Kilcoyne Ji-Yong Julie Kim Missy Lavender Erica E Marsh Kristen A Matteson Jacqueline A Maybin Christine N Metz Inmaculada Moreno Kami Silk Marni Sommer Carlos Simon Ridhi Tariyal Hugh S Taylor Günter P Wagner Linda G Griffith

Am J Obstet Gynecol 2020 11 21;223(5):624-664. Epub 2020 Jul 21.

Center for Gynepathology Research, Massachusetts Institute of Technology, Cambridge, MA.

Women's health concerns are generally underrepresented in basic and translational research, but reproductive health in particular has been hampered by a lack of understanding of basic uterine and menstrual physiology. Menstrual health is an integral part of overall health because between menarche and menopause, most women menstruate. Yet for tens of millions of women around the world, menstruation regularly and often catastrophically disrupts their physical, mental, and social well-being. Enhancing our understanding of the underlying phenomena involved in menstruation, abnormal uterine bleeding, and other menstruation-related disorders will move us closer to the goal of personalized care. Furthermore, a deeper mechanistic understanding of menstruation-a fast, scarless healing process in healthy individuals-will likely yield insights into a myriad of other diseases involving regulation of vascular function locally and systemically. We also recognize that many women now delay pregnancy and that there is an increasing desire for fertility and uterine preservation. In September 2018, the Gynecologic Health and Disease Branch of the Eunice Kennedy Shriver National Institute of Child Health and Human Development convened a 2-day meeting, "Menstruation: Science and Society" with an aim to "identify gaps and opportunities in menstruation science and to raise awareness of the need for more research in this field." Experts in fields ranging from the evolutionary role of menstruation to basic endometrial biology (including omic analysis of the endometrium, stem cells and tissue engineering of the endometrium, endometrial microbiome, and abnormal uterine bleeding and fibroids) and translational medicine (imaging and sampling modalities, patient-focused analysis of menstrual disorders including abnormal uterine bleeding, smart technologies or applications and mobile health platforms) to societal challenges in health literacy and dissemination frameworks across different economic and cultural landscapes shared current state-of-the-art and future vision, incorporating the patient voice at the launch of the meeting. Here, we provide an enhanced meeting report with extensive up-to-date (as of submission) context, capturing the spectrum from how the basic processes of menstruation commence in response to progesterone withdrawal, through the role of tissue-resident and circulating stem and progenitor cells in monthly regeneration-and current gaps in knowledge on how dysregulation leads to abnormal uterine bleeding and other menstruation-related disorders such as adenomyosis, endometriosis, and fibroids-to the clinical challenges in diagnostics, treatment, and patient and societal education. We conclude with an overview of how the global agenda concerning menstruation, and specifically menstrual health and hygiene, are gaining momentum, ranging from increasing investment in addressing menstruation-related barriers facing girls in schools in low- to middle-income countries to the more recent "menstrual equity" and "period poverty" movements spreading across high-income countries.
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http://dx.doi.org/10.1016/j.ajog.2020.06.004DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7661839PMC
November 2020

Circulating TNF-like protein 1A (TL1A) is elevated early in rheumatoid arthritis and depends on TNF.

Authors:
Yun-Jeong Song In Ah Choi Françoise Meylan M Kristen Demoruelle Taylor Farley Arianne C Richard Eric Hawley John Botson Yoo Jin Hong Eun Young Lee Sabina R Mian Bartlett C Hamilton Geoffrey M Thiele Ted R Mikuls Naveen Gara Chris D Ward Sarah Lamberth Kevin D Deane Theo Heller Michael M Ward David M Lee Thi-Sau Migone William Stohl James R O'Dell Jill M Norris V Michael Holers Peter Gregersen Yeong-Wook Song Richard M Siegel

Arthritis Res Ther 2020 05 7;22(1):106. Epub 2020 May 7.

Immunoregulation Section, Autoimmunity Branch, NIAMS, NIH, Bethesda, MD, USA.

Background: The tumor necrosis factor (TNF) superfamily cytokine TNF-like protein 1A (TL1A) and its receptor DR3 are essential for diverse animal models of autoimmune disease and may be pathogenic in rheumatoid arthritis (RA). However, the relationship of TL1A to disease duration, activity, and response to anti-TNF and other therapies in RA is not clear.

Methods: We measured soluble TL1A in synovial fluid (SF), serum, or plasma from RA first-degree relatives (FDRs) and in early RA and established disease. We measured the effects of anti-TNF and methotrexate (MTX) therapy on circulating TL1A from multiple independent RA treatment trials. We also determined the ability of a blocking anti-TL1A antibody to inhibit clinical disease and articular bone destruction in the murine collagen-induced arthritis (CIA) model of human RA.

Results: Soluble TL1A was specifically elevated in the blood and SF of patients with RA compared to patients with other diseases and was elevated early in disease and in at-risk anti-cyclic citrullinated peptide (CCP) (+) first-degree relatives (FDRs). Therapeutic TNF inhibition reduced serum TL1A in both responders and non-responders, whereas TL1A declined following MTX treatment only in responders. In murine CIA, TL1A blockade was clinically efficacious and reduced bone erosions.

Conclusions: TL1A is specifically elevated in RA from early in the disease course and in at-risk FDRs. The decline in TL1A after TNF blockade suggests that TL1A levels may be a useful biomarker for TNF activity in RA. These results support the further investigation of the relationship between TL1A and TNF and TL1A blockade as a potential therapeutic strategy in RA.
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http://dx.doi.org/10.1186/s13075-020-02198-9DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7204024PMC
May 2020

Allele-specific expression changes dynamically during T cell activation in HLA and other autoimmune loci.

Authors:
Maria Gutierrez-Arcelus Yuriy Baglaenko Jatin Arora Susan Hannes Yang Luo Tiffany Amariuta Nikola Teslovich Deepak A Rao Joerg Ermann A Helena Jonsson Cristina Navarrete Stephen S Rich Kent D Taylor Jerome I Rotter Peter K Gregersen Tonu Esko Michael B Brenner Soumya Raychaudhuri

Nat Genet 2020 03 17;52(3):247-253. Epub 2020 Feb 17.

Center for Data Sciences, Brigham and Women's Hospital, Harvard Medical School, Boston, MA, USA.

Genetic studies have revealed that autoimmune susceptibility variants are over-represented in memory CD4 T cell regulatory elements. Understanding how genetic variation affects gene expression in different T cell physiological states is essential for deciphering genetic mechanisms of autoimmunity. Here, we characterized the dynamics of genetic regulatory effects at eight time points during memory CD4 T cell activation with high-depth RNA-seq in healthy individuals. We discovered widespread, dynamic allele-specific expression across the genome, where the balance of alleles changes over time. These genes were enriched fourfold within autoimmune loci. We found pervasive dynamic regulatory effects within six HLA genes. HLA-DQB1 alleles had one of three distinct transcriptional regulatory programs. Using CRISPR-Cas9 genomic editing we demonstrated that a promoter variant is causal for T cell-specific control of HLA-DQB1 expression. Our study shows that genetic variation in cis-regulatory elements affects gene expression in a manner dependent on lymphocyte activation status, contributing to the interindividual complexity of immune responses.
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http://dx.doi.org/10.1038/s41588-020-0579-4DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7135372PMC
March 2020

IRF5 genetic risk variants drive myeloid-specific IRF5 hyperactivation and presymptomatic SLE.

Authors:
Dan Li Bharati Matta Su Song Victoria Nelson Kirsten Diggins Kim R Simpfendorfer Peter K Gregersen Peter Linsley Betsy J Barnes

JCI Insight 2020 01 30;5(2). Epub 2020 Jan 30.

Center for Autoimmune, Musculoskeletal and Hematopoietic Diseases, The Feinstein Institutes for Medical Research, Manhasset, New York, USA.

Genetic variants within or near the interferon regulatory factor 5 (IRF5) locus associate with systemic lupus erythematosus (SLE) across ancestral groups. The major IRF5-SLE risk haplotype is common across populations, yet immune functions for the risk haplotype are undefined. We characterized the global immune phenotype of healthy donors homozygous for the major risk and nonrisk haplotypes and identified cell lineage-specific alterations that mimic presymptomatic SLE. Contrary to previous studies in B lymphoblastoid cell lines and SLE immune cells, IRF5 genetic variants had little effect on IRF5 protein levels in healthy donors. Instead, we detected basal IRF5 hyperactivation in the myeloid compartment of risk donors that drives the SLE immune phenotype. Risk donors were anti-nuclear antibody positive with anti-Ro and -MPO specificity, had increased circulating plasma cells and plasmacytoid dendritic cells, and had enhanced spontaneous NETosis. The IRF5-SLE immune phenotype was conserved over time and probed mechanistically by ex vivo coculture, indicating that risk neutrophils are drivers of the global immune phenotype. RNA-Seq of risk neutrophils revealed increased IRF5 transcript expression, IFN pathway enrichment, and decreased expression of ROS pathway genes. Altogether, the data support that individuals carrying the IRF5-SLE risk haplotype are more susceptible to environmental/stochastic influences that trigger chronic immune activation, predisposing to the development of clinical SLE.
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http://dx.doi.org/10.1172/jci.insight.124020DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7098722PMC
January 2020

Perceived Stress and Inflammatory Arthritis: A Prospective Investigation in the Studies of the Etiologies of Rheumatoid Arthritis Cohort.

Authors:
Kristen J Polinski Elizabeth A Bemis Marie Feser Jennifer Seifert M Kristen Demoruelle Christopher C Striebich Stacey Brake James R O'Dell Ted R Mikuls Michael H Weisman Peter K Gregersen Richard M Keating Jane Buckner Perry Nicassio V Michael Holers Kevin D Deane Jill M Norris

Arthritis Care Res (Hoboken) 2020 12 6;72(12):1766-1771. Epub 2020 Nov 6.

Colorado School of Public Health, Aurora.

Objective: The aim of this study was to determine the association of perceived stress with incident inflammatory arthritis (IA) defined as having at least 1 joint consistent with rheumatoid arthritis (RA)-like synovitis based on examination.

Methods: We conducted a prospective cohort study in the Studies of the Etiologies of Rheumatoid Arthritis cohort. Participants without IA were recruited if they were a first-degree relative of an RA proband or screened positive for anti-citrullinated protein antibody. Perceived stress was measured using the Perceived Stress Scale-14 (PSS-14), in which scores can range from 0 to 56, and a higher score indicates greater perceived stress. The total PSS-14 score, as well as 2 subscores indicative of perceived distress and self-efficacy, were averaged across all study visits until development of IA or the last follow-up. Hazard ratios (HRs) and 95% confidence intervals (95% CIs) of IA associated with average PSS-14 scores were obtained using Cox proportional hazards models.

Results: The mean total PSS-14 score was 20.4. We found that a 1-point increase in the perceived distress score was significantly associated with a 10-percent increase in the risk of IA (adjusted HR 1.10 [95% CI 1.02-1.19]). Total PSS-14 and self-efficacy were not associated with IA risk (adjusted HR 1.05 [95% CI 0.99-1.10] and 1.04 [95% CI 0.91-1.18], respectively).

Conclusion: An association between perceived distress and incident IA was observed in this at-risk cohort. Replication of this finding in other preclinical and at-risk RA populations is needed.
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http://dx.doi.org/10.1002/acr.24085DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7145743PMC
December 2020

Genetic associations with radiological damage in rheumatoid arthritis: Meta-analysis of seven genome-wide association studies of 2,775 cases.

Authors:
Matthew Traylor Rachel Knevel Jing Cui John Taylor Westra Harm-Jan Philip G Conaghan Andrew P Cope Charles Curtis Paul Emery Stephen Newhouse Hamel Patel Sophia Steer Peter Gregersen Nancy A Shadick Michael E Weinblatt Annette Van Der Helm-van Mil Jennifer H Barrett Ann W Morgan Cathryn M Lewis Ian C Scott

PLoS One 2019 9;14(10):e0223246. Epub 2019 Oct 9.

Primary Care Centre Versus Arthritis, Research Institute for Primary Care and Health Sciences, Primary Care Sciences, Keele University, Keele, United Kingdom.

Background: Previous studies of radiological damage in rheumatoid arthritis (RA) have used candidate-gene approaches, or evaluated single genome-wide association studies (GWAS). We undertook the first meta-analysis of GWAS of RA radiological damage to: (1) identify novel genetic loci for this trait; and (2) test previously validated variants.

Methods: Seven GWAS (2,775 RA cases, of a range of ancestries) were combined in a meta-analysis. Radiological damage was assessed using modified Larsen scores, Sharp van Der Heijde scores, and erosive status. Single nucleotide polymophsim (SNP) associations with radiological damage were tested at a single time-point using regression models. Primary analyses included age and disease duration as covariates. Secondary analyses also included rheumatoid factor (RF). Meta-analyses were undertaken in trans-ethnic and European-only cases.

Results: In the trans-ethnic primary meta-analysis, one SNP (rs112112734) in close proximity to HLA-DRB1, and strong linkage disequilibrium with the shared-epitope, attained genome-wide significance (P = 4.2x10-8). In the secondary analysis (adjusting for RF) the association was less significant (P = 1.7x10-6). In both trans-ethnic primary and secondary meta-analyses 14 regions contained SNPs with associations reaching P<5x10-6; in the European primary and secondary analyses 13 and 10 regions contained SNPs reaching P<5x10-6, respectively. Of the previously validated SNPs for radiological progression, only rs660895 (tagging HLA-DRB1*04:01) attained significance (P = 1.6x10-5) and had a consistent direction of effect across GWAS.

Conclusions: Our meta-analysis confirms the known association between the HLA-DRB1 shared epitope and RA radiological damage. The lack of replication of previously validated non-HLA markers highlights a requirement for further research to deliver clinically-useful prognostic genetic markers.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0223246PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6785117PMC
March 2020

Distinct HLA Associations with Rheumatoid Arthritis Subsets Defined by Serological Subphenotype.

Authors:
Chikashi Terao Boel Brynedal Zuomei Chen Xia Jiang Helga Westerlind Monika Hansson Per-Johan Jakobsson Karin Lundberg Karl Skriner Guy Serre Johan Rönnelid Linda Mathsson-Alm Mikael Brink Solbritt Rantapää Dahlqvist Leonid Padyukov Peter K Gregersen Anne Barton Lars Alfredsson Lars Klareskog Soumya Raychaudhuri

Am J Hum Genet 2019 Oct;105(4):880

Division of Rheumatology, Immunology, and Allergy Brigham and Women's Hospital, Harvard Medical School, Boston, MA 02115, USA; Division of Genetics, Brigham and Women's Hospital, Harvard Medical School, Boston, MA 02115, USA; Program in Medical and Population Genetics, Broad Institute, Cambridge, MA 02142, USA; Arthritis Research UK Centre for Genetics and Genomics, Centre for Musculoskeletal Research, Faculty of Biology, Medicine and Health, Manchester Academic Health Science Centre, The University of Manchester, Manchester, Oxford Road, Manchester M13 9PT, UK; Center for Data Sciences, Brigham and Women's Hospital, Harvard Medical School, Boston, MA 02115, USA. Electronic address:

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http://dx.doi.org/10.1016/j.ajhg.2019.09.018DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6817550PMC
October 2019

Distinct HLA Associations with Rheumatoid Arthritis Subsets Defined by Serological Subphenotype.

Authors:
Chikashi Terao Boel Brynedal Zuomei Chen Xia Jiang Helga Westerlind Monika Hansson Per-Johan Jakobsson Karin Lundberg Karl Skriner Guy Serre Johan Rönnelid Linda Mathsson-Alm Mikael Brink Solbritt Rantapää Dahlqvist Leonid Padyukov Peter K Gregersen Anne Barton Lars Alfredsson Lars Klareskog Soumya Raychaudhuri

Am J Hum Genet 2019 09 29;105(3):616-624. Epub 2019 Aug 29.

Division of Rheumatology, Inflammation, and Immunity, Brigham and Women's Hospital, Harvard Medical School, Boston, MA 02115, USA; Division of Genetics, Brigham and Women's Hospital, Harvard Medical School, Boston, MA 02115, USA; Program in Medical and Population Genetics, Broad Institute, Cambridge, MA 02142, USA; Versus Arthritis Centre for Genetics and Genomics, Centre for Musculoskeletal Research, Faculty of Biology, Medicine and Health, Manchester Academic Health Science Centre, The University of Manchester, Manchester, Oxford Road, Manchester M13 9PT, UK; Center for Data Sciences, Brigham and Women's Hospital, Harvard Medical School, Boston, MA 02115, USA. Electronic address:

Rheumatoid arthritis (RA) is the most common immune-mediated arthritis. Anti-citrullinated peptide antibodies (ACPA) are highly specific to RA and assayed with the commercial CCP2 assay. Genetic drivers of RA within the MHC are different for CCP2-positive and -negative subsets of RA, particularly at HLA-DRB1. However, aspartic acid at amino acid position 9 in HLA-B (B) increases risk to both RA subsets. Here we explore how individual serologies associated with RA drive associations within the MHC. To define MHC differences for specific ACPA serologies, we quantified a total of 19 separate ACPAs in RA-affected case subjects from four cohorts (n = 6,805). We found a cluster of tightly co-occurring antibodies (canonical serologies, containing CCP2), along with several independently expressed antibodies (non-canonical serologies). After imputing HLA variants into 6,805 case subjects and 13,467 control subjects, we tested associations between the HLA region and RA subgroups based on the presence of canonical and/or non-canonical serologies. We examined CCP2(+) and CCP2(-) RA-affected case subjects separately. In CCP2(-) RA, we observed that the association between CCP2(-) RA and B was derived from individuals who were positive for non-canonical serologies (omnibus_p = 9.2 × 10). Similarly, we observed in CCP2(+) RA that associations between subsets of CCP2(+) RA and B were negatively correlated with the number of positive canonical serologies (p = 0.0096). These findings suggest unique genetic characteristics underlying fine-specific ACPAs, suggesting that RA may be further subdivided beyond simply seropositive and seronegative.
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http://dx.doi.org/10.1016/j.ajhg.2019.08.002DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6731376PMC
September 2019

Focused HLA analysis in Caucasians with myositis identifies significant associations with autoantibody subgroups.

Authors:
Simon Rothwell Hector Chinoy Janine A Lamb Frederick W Miller Lisa G Rider Lucy R Wedderburn Neil J McHugh Andrew L Mammen Zoe E Betteridge Sarah L Tansley John Bowes Jiří Vencovský Claire T Deakin Katalin Dankó Limaye Vidya Albert Selva-O'Callaghan Lauren M Pachman Ann M Reed Øyvind Molberg Olivier Benveniste Pernille R Mathiesen Timothy R D J Radstake Andrea Doria Jan de Bleecker Annette T Lee Michael G Hanna Pedro M Machado William E Ollier Peter K Gregersen Leonid Padyukov Terrance P O'Hanlon Robert G Cooper Ingrid E Lundberg

Ann Rheum Dis 2019 07 28;78(7):996-1002. Epub 2019 May 28.

Division of Rheumatology, Department of Medicine, Karolinska University Hospital, Stockholm, Sweden.

Objectives: Idiopathic inflammatory myopathies (IIM) are a spectrum of rare autoimmune diseases characterised clinically by muscle weakness and heterogeneous systemic organ involvement. The strongest genetic risk is within the major histocompatibility complex (MHC). Since autoantibody presence defines specific clinical subgroups of IIM, we aimed to correlate serotype and genotype, to identify novel risk variants in the MHC region that co-occur with IIM autoantibodies.

Methods: We collected available autoantibody data in our cohort of 2582 Caucasian patients with IIM. High resolution human leucocyte antigen (HLA) alleles and corresponding amino acid sequences were imputed using SNP2HLA from existing genotyping data and tested for association with 12 autoantibody subgroups.

Results: We report associations with eight autoantibodies reaching our study-wide significance level of p<2.9×10. Associations with the 8.1 ancestral haplotype were found with anti-Jo-1 (HLA-B*08:01, p=2.28×10 and HLA-DRB1*03:01, p=3.25×10), anti-PM/Scl (HLA-DQB1*02:01, p=1.47×10) and anti-cN1A autoantibodies (HLA-DRB1*03:01, p=1.40×10). Associations independent of this haplotype were found with anti-Mi-2 (HLA-DRB1*07:01, p=4.92×10) and anti-HMGCR autoantibodies (HLA-DRB1*11, p=5.09×10). Amino acid positions may be more strongly associated than classical HLA associations; for example with anti-Jo-1 autoantibodies and position 74 of HLA-DRB1 (p=3.47×10) and position 9 of HLA-B (p=7.03×10). We report novel genetic associations with HLA-DQB1 anti-TIF1 autoantibodies and identify haplotypes that may differ between adult-onset and juvenile-onset patients with these autoantibodies.

Conclusions: These findings provide new insights regarding the functional consequences of genetic polymorphisms within the MHC. As autoantibodies in IIM correlate with specific clinical features of disease, understanding genetic risk underlying development of autoantibody profiles has implications for future research.
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http://dx.doi.org/10.1136/annrheumdis-2019-215046DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6585280PMC
July 2019

Defining inflammatory cell states in rheumatoid arthritis joint synovial tissues by integrating single-cell transcriptomics and mass cytometry.

Authors:
Fan Zhang Kevin Wei Kamil Slowikowski Chamith Y Fonseka Deepak A Rao Stephen Kelly Susan M Goodman Darren Tabechian Laura B Hughes Karen Salomon-Escoto Gerald F M Watts A Helena Jonsson Javier Rangel-Moreno Nida Meednu Cristina Rozo William Apruzzese Thomas M Eisenhaure David J Lieb David L Boyle Arthur M Mandelin Brendan F Boyce Edward DiCarlo Ellen M Gravallese Peter K Gregersen Larry Moreland Gary S Firestein Nir Hacohen Chad Nusbaum James A Lederer Harris Perlman Costantino Pitzalis Andrew Filer V Michael Holers Vivian P Bykerk Laura T Donlin Jennifer H Anolik Michael B Brenner Soumya Raychaudhuri

Nat Immunol 2019 07 6;20(7):928-942. Epub 2019 May 6.

Center for Data Sciences, Brigham and Women's Hospital, Boston, MA, USA.

To define the cell populations that drive joint inflammation in rheumatoid arthritis (RA), we applied single-cell RNA sequencing (scRNA-seq), mass cytometry, bulk RNA sequencing (RNA-seq) and flow cytometry to T cells, B cells, monocytes, and fibroblasts from 51 samples of synovial tissue from patients with RA or osteoarthritis (OA). Utilizing an integrated strategy based on canonical correlation analysis of 5,265 scRNA-seq profiles, we identified 18 unique cell populations. Combining mass cytometry and transcriptomics revealed cell states expanded in RA synovia: THY1(CD90)HLA-DRA sublining fibroblasts, IL1B pro-inflammatory monocytes, ITGAXTBX21 autoimmune-associated B cells and PDCD1 peripheral helper T (T) cells and follicular helper T (T) cells. We defined distinct subsets of CD8 T cells characterized by GZMK, GZMB, and GNLY phenotypes. We mapped inflammatory mediators to their source cell populations; for example, we attributed IL6 expression to THY1HLA-DRA fibroblasts and IL1B production to pro-inflammatory monocytes. These populations are potentially key mediators of RA pathogenesis.
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http://dx.doi.org/10.1038/s41590-019-0378-1DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6602051PMC
July 2019

Influence of genetic copy number variants of the human GLUT3 glucose transporter gene on protein expression, glycolysis and rheumatoid arthritis risk: A genetic replication study.

Authors:
Kim R Simpfendorfer Wentian Li Andrew Shih Hongxiu Wen Harini P Kothari Edward A Einsidler Arthur Wuster Julie Hunkapiller Timothy W Behrens Robert R Graham Michael J Townsend Doron M Behar Rui Hu Elliott Greenspan Peter K Gregersen

Mol Genet Metab Rep 2019 Jun 6;19:100470. Epub 2019 Apr 6.

Robert S. Boas Center for Genomics and Human Genetics, the Feinstein Institute for Medical Research, 350 Community Drive, Manhasset, New York, USA.

Objectives: The gene encoding glucose transporter 3 (GLUT3, ) is present in the human population at variable copy number. An overt disease phenotype of copy number variants has not been reported; however, deletion of has been previously reported to protect carriers from rheumatoid arthritis, implicating GLUT3 as a therapeutic target in rheumatoid arthritis. Here we aim to perform functional analysis of GLUT3 copy number variants in immune cells, and test the reported protective association of the GLUT3 copy number variants for rheumatoid arthritis in a genetic replication study.

Methods: Cells from genotyped healthy controls were analyzed for /GLUT3 expression and glycolysis capacity. We genotyped the copy number variant in four independent cohorts of rheumatoid arthritis and controls and one cohort of multiple sclerosis and controls.

Results: Heterozygous deletion of correlates directly with expression levels of GLUT3 and influences glycolysis rates in the human immune system. The frequency of the copy number variant is not different between rheumatoid arthritis, multiple sclerosis and control groups.

Conclusions: Despite a robust gene copy number dependent phenotype, our study of large groups of rheumatoid arthritis cases and controls provides no evidence for rheumatoid arthritis disease protection in deletion carriers. These data emphasize the importance of well powered replication studies to confirm or refute genetic associations, particularly for relatively rare variants.
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http://dx.doi.org/10.1016/j.ymgmr.2019.100470DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6453668PMC
June 2019

Anticyclic Citrullinated Peptide Antibodies 3.1 and Anti-CCP-IgA Are Associated with Increasing Age in Individuals Without Rheumatoid Arthritis.

Authors:
Heather M Berens Kristen J Polinski Ted R Mikuls Sonia Khatter Justin August Ashley Visser Michael Mahler Michael H Weisman James R O'Dell Richard M Keating Jane H Buckner Peter K Gregersen Jill M Norris V Michael Holers Kevin D Deane M Kristen Demoruelle

J Rheumatol 2019 12 15;46(12):1556-1559. Epub 2019 Apr 15.

From the Division of Rheumatology, University of Colorado Anschutz Medical Campus, Aurora; Department of Epidemiology, Colorado School of Public Health, Aurora, Colorado; Division of Rheumatology, University of Nebraska Medical Center, Omaha; Division of Rheumatology, Veterans Affairs (VA) Nebraska Western Iowa Health Care System, Omaha, Nebraska; Department of Research, Inova Diagnostics, San Diego; Division of Rheumatology, Cedars-Sinai Medical Center, Los Angeles; Division of Rheumatology, Scripps Green Hospital, La Jolla, California; Department of Immunology, Benaroya Research Institute at Virginia Mason, Seattle, Washington; Center for Genomics and Human Genetics, Feinstein Institute for Medical Research, Manhasset, New York, USA.

Objective: We investigated the association of age and anticyclic citrullinated peptide antibodies (anti-CCP) in subjects without rheumatoid arthritis (RA).

Methods: Serum was tested for anti-CCP3.1 (IgG/IgA) in 678 first-degree relatives (FDR) of patients with RA and 330 patients with osteoarthritis (OA). Individual isotypes (anti-CCP-IgA and anti-CCP-IgG) were also tested in all FDR.

Results: In FDR, increasing age was significantly associated with positivity for anti-CCP3.1 (per year, OR 1.03) and anti-CCP-IgA (per year, OR 1.05) but not anti-CCP-IgG. In FDR and OA subjects, anti-CCP3.1 prevalence was significantly increased after age 50 years.

Conclusion: Increasing age in individuals without RA should be considered in the interpretation of anti-CCP3.1 positivity.
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http://dx.doi.org/10.3899/jrheum.180897DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6800826PMC
December 2019

Type 1 Diabetes Risk in African-Ancestry Participants and Utility of an Ancestry-Specific Genetic Risk Score.

Authors:
Suna Onengut-Gumuscu Wei-Min Chen Catherine C Robertson Jessica K Bonnie Emily Farber Zhennan Zhu Jorge R Oksenberg Steven R Brant S Louis Bridges Jeffrey C Edberg Robert P Kimberly Peter K Gregersen Marian J Rewers Andrea K Steck Mary H Black Dana Dabelea Catherine Pihoker Mark A Atkinson Lynne E Wagenknecht Jasmin Divers Ronny A Bell Henry A Erlich Patrick Concannon Stephen S Rich

Diabetes Care 2019 03 18;42(3):406-415. Epub 2019 Jan 18.

Center for Public Health Genomics, University of Virginia, Charlottesville, VA

Objective: Genetic risk scores (GRS) have been developed that differentiate individuals with type 1 diabetes from those with other forms of diabetes and are starting to be used for population screening; however, most studies were conducted in European-ancestry populations. This study identifies novel genetic variants associated with type 1 diabetes risk in African-ancestry participants and develops an African-specific GRS.

Research Design And Methods: We generated single nucleotide polymorphism (SNP) data with the ImmunoChip on 1,021 African-ancestry participants with type 1 diabetes and 2,928 control participants. HLA class I and class II alleles were imputed using SNP2HLA. Logistic regression models were used to identify genome-wide significant ( < 5.0 × 10) SNPs associated with type 1 diabetes in the African-ancestry samples and validate SNPs associated with risk in known European-ancestry loci ( < 2.79 × 10).

Results: African-specific (HLA-*03:01-HLA-*02:01) and known European-ancestry HLA haplotypes (HLA-*03:01-HLA-*05:01-HLA-*02:01, HLA-*04:01-HLA-*03:01-HLA-*03:02) were significantly associated with type 1 diabetes risk. Among European-ancestry defined non-HLA risk loci, six risk loci were significantly associated with type 1 diabetes in subjects of African ancestry. An African-specific GRS provided strong prediction of type 1 diabetes risk (area under the curve 0.871), performing significantly better than a European-based GRS and two polygenic risk scores in independent discovery and validation cohorts.

Conclusions: Genetic risk of type 1 diabetes includes ancestry-specific, disease-associated variants. The GRS developed here provides improved prediction of type 1 diabetes in African-ancestry subjects and a means to identify groups of individuals who would benefit from immune monitoring for early detection of islet autoimmunity.
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http://dx.doi.org/10.2337/dc18-1727DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6385701PMC
March 2019

Genome-wide meta-analysis reveals shared new in systemic seropositive rheumatic diseases.

Authors:
Marialbert Acosta-Herrera Martin Kerick David González-Serna Cisca Wijmenga Andre Franke Peter K Gregersen Leonid Padyukov Jane Worthington Timothy James Vyse Marta Eugenia Alarcón-Riquelme Maureen D Mayes Javier Martin

Ann Rheum Dis 2019 03 20;78(3):311-319. Epub 2018 Dec 20.

Institute of Parasitology and Biomedicine López-Neyra, IPBLN-CSIC, PTS Granada, Granada, Spain

Objective: Immune-mediated inflammatory diseases (IMIDs) are heterogeneous and complex conditions with overlapping clinical symptoms and elevated familial aggregation, which suggests the existence of a shared genetic component. In order to identify this genetic background in a systematic fashion, we performed the first cross-disease genome-wide meta-analysis in systemic seropositive rheumatic diseases, namely, systemic sclerosis, systemic lupus erythematosus, rheumatoid arthritis and idiopathic inflammatory myopathies.

Methods: We meta-analysed ~6.5 million single nucleotide polymorphisms in 11 678 cases and 19 704 non-affected controls of European descent populations. The functional roles of the associated variants were interrogated using publicly available databases.

Results: Our analysis revealed five shared genome-wide significant independent that had not been previously associated with these diseases: , , , and . All of these are related with immune processes such as interferon and epidermal growth factor signalling, response to methotrexate, cytoskeleton dynamics and coagulation cascade. Remarkably, several of the associated are known key players in autoimmunity, which supports the validity of our results. All the associated variants showed significant functional enrichment in DNase hypersensitivity sites, chromatin states and histone marks in relevant immune cells, including shared expression quantitative trait . Additionally, our results were significantly enriched in drugs that are being tested for the treatment of the diseases under study.

Conclusions: We have identified shared new risk with functional value across diseases and pinpoint new potential candidate that could be further investigated. Our results highlight the potential of drug repositioning among related systemic seropositive rheumatic IMIDs.
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http://dx.doi.org/10.1136/annrheumdis-2018-214127DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6800208PMC
March 2019

Correction: Identification of grade and origin specific cell populations in serous epithelial ovarian cancer by single cell RNA-seq.

Authors:
Andrew J Shih Andrew Menzin Jill Whyte John Lovecchio Anthony Liew Houman Khalili Tawfiqul Bhuiya Peter K Gregersen Annette T Lee

PLoS One 2018 4;13(12):e0208778. Epub 2018 Dec 4.

[This corrects the article DOI: 10.1371/journal.pone.0206785.].
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0208778PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6279024PMC
December 2018

Genetic influences on susceptibility to rheumatoid arthritis in African-Americans.

Authors:
Vincent A Laufer Hemant K Tiwari Richard J Reynolds Maria I Danila Jelai Wang Jeffrey C Edberg Robert P Kimberly Leah C Kottyan John B Harley Ted R Mikuls Peter K Gregersen Devin M Absher Carl D Langefeld Donna K Arnett S Louis Bridges

Hum Mol Genet 2019 03;28(5):858-874

Department of Medicine, University of Alabama at Birmingham, Birmingham, AL, USA.

Large meta-analyses of rheumatoid arthritis (RA) susceptibility in European (EUR) and East Asian (EAS) populations have identified >100 RA risk loci, but genome-wide studies of RA in African-Americans (AAs) are absent. To address this disparity, we performed an analysis of 916 AA RA patients and 1392 controls and aggregated our data with genotyping data from >100 000 EUR and Asian RA patients and controls. We identified two novel risk loci that appear to be specific to AAs: GPC5 and RBFOX1 (PAA < 5 × 10-9). Most RA risk loci are shared across different ethnicities, but among discordant loci, we observed strong enrichment of variants having large effect sizes. We found strong evidence of effect concordance for only 3 of the 21 largest effect index variants in EURs. We used the trans-ethnic fine-mapping algorithm PAINTOR3 to prioritize risk variants in >90 RA risk loci. Addition of AA data to those of EUR and EAS descent enabled identification of seven novel high-confidence candidate pathogenic variants (defined by posterior probability > 0.8). In summary, our trans-ethnic analyses are the first to include AAs, identified several new RA risk loci and point to candidate pathogenic variants that may underlie this common autoimmune disease. These findings may lead to better ways to diagnose or stratify treatment approaches in RA.
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http://dx.doi.org/10.1093/hmg/ddy395DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6381313PMC
March 2019

Identification of grade and origin specific cell populations in serous epithelial ovarian cancer by single cell RNA-seq.

Authors:
Andrew J Shih Andrew Menzin Jill Whyte John Lovecchio Anthony Liew Houman Khalili Tawfiqul Bhuiya Peter K Gregersen Annette T Lee

PLoS One 2018 1;13(11):e0206785. Epub 2018 Nov 1.

Robert S. Boas Center for Genomics and Human Genetics, The Feinstein Institute for Medical Research, Manhasset, New York, United States of America.

Here we investigated different cell populations within ovarian cancer using single-cell RNA seq: fourteen samples from nine patients with differing grades (high grade, low grade and benign) as well as different origin sites (primary and metastatic tumor site, ovarian in origin and fallopian in origin). We were able to identify sixteen distinct cell populations with specific cells correlated to high grade tumors, low grade tumors, benign and one population unique to a patient with a breast cancer relapse. Furthermore the proportion of these populations changes from primary to metastatic in a shift from mainly epithelial cells to leukocytes with few cancer epithelial cells in the metastases. Differential gene expression shows myeloid lineage cells are the primary cell group expressing soluble factors in primary samples while fibroblasts do so in metastatic samples. The leukocytes that were captured did not seem to be suppressed through known pro-tumor cytokines from any of the cell populations. Single cell RNA-seq is necessary to de-tangle cellular heterogeneity for better understanding of ovarian cancer progression.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0206785PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6211742PMC
April 2019

Breathing New Life into Interstitial Lung Disease in Rheumatoid Arthritis.

Authors:
Peter K Gregersen Ellen M Gravallese

N Engl J Med 2018 12 20;379(23):2265-2266. Epub 2018 Oct 20.

From the Feinstein Institute for Medical Research, Robert S. Boas Center for Genomics and Human Genetics, Manhasset, NY (P.K.G.); and the Department of Medicine, Division of Rheumatology, University of Massachusetts Medical School, Worcester (E.M.G.).

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http://dx.doi.org/10.1056/NEJMe1811767DOI Listing
December 2018

Fine-mapping and functional studies highlight potential causal variants for rheumatoid arthritis and type 1 diabetes.

Authors:
Harm-Jan Westra Marta Martínez-Bonet Suna Onengut-Gumuscu Annette Lee Yang Luo Nikola Teslovich Jane Worthington Javier Martin Tom Huizinga Lars Klareskog Solbritt Rantapaa-Dahlqvist Wei-Min Chen Aaron Quinlan John A Todd Steve Eyre Peter A Nigrovic Peter K Gregersen Stephen S Rich Soumya Raychaudhuri

Nat Genet 2018 10 17;50(10):1366-1374. Epub 2018 Sep 17.

Center for Data Sciences, Brigham and Women's Hospital, Harvard Medical School, Boston, MA, USA.

To define potentially causal variants for autoimmune disease, we fine-mapped 76 rheumatoid arthritis (11,475 cases, 15,870 controls) and type 1 diabetes loci (9,334 cases, 11,111 controls). After sequencing 799 1-kilobase regulatory (H3K4me3) regions within these loci in 568 individuals, we observed accurate imputation for 89% of common variants. We defined credible sets of ≤5 causal variants at 5 rheumatoid arthritis and 10 type 1 diabetes loci. We identified potentially causal missense variants at DNASE1L3, PTPN22, SH2B3, and TYK2, and noncoding variants at MEG3, CD28-CTLA4, and IL2RA. We also identified potential candidate causal variants at SIRPG and TNFAIP3. Using functional assays, we confirmed allele-specific protein binding and differential enhancer activity for three variants: the CD28-CTLA4 rs117701653 SNP, MEG3 rs34552516 indel, and TNFAIP3 rs35926684 indel.
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http://dx.doi.org/10.1038/s41588-018-0216-7DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6364548PMC
October 2018

Serologic features of cohorts with variable genetic risk for systemic lupus erythematosus.

Authors:
Jyotsna Bhattacharya Karalyn Pappas Bahtiyar Toz Cynthia Aranow Meggan Mackay Peter K Gregersen Ogobara Doumbo Abdel Kader Traore Martin L Lesser Maureen McMahon Tammy Utset Earl Silverman Deborah Levy William J McCune Meenakshi Jolly Daniel Wallace Michael Weisman Juanita Romero-Diaz Betty Diamond

Mol Med 2018 06 1;24(1):24. Epub 2018 Jun 1.

The Feinstein Institute for Medical Research, Center for Autoimmune, Musculoskeletal and Hematopoietic Diseases, 350 Community Dr, Manhasset, NY, 11030, USA.

Background: Systemic lupus erythematosus (SLE) is an autoimmune disease with genetic, hormonal, and environmental influences. In Western Europe and North America, individuals of West African descent have a 3-4 fold greater incidence of SLE than Caucasians. Paradoxically, West Africans in sub-Saharan Africa appear to have a low incidence of SLE, and some studies suggest a milder disease with less nephritis. In this study, we analyzed sera from African American female SLE patients and four other cohorts, one with SLE and others with varying degrees of risk for SLE in order to identify serologic factors that might correlate with risk of or protection against SLE.

Methods: Our cohorts included West African women with previous malaria infection assumed to be protected from development of SLE, clinically unaffected sisters of SLE patients with high risk of developing SLE, healthy African American women with intermediate risk, healthy Caucasian women with low risk of developing SLE, and women with a diagnosis of SLE. We developed a lupus risk index (LRI) based on titers of IgM and IgG anti-double stranded DNA antibodies and levels of C1q.

Results: The risk index was highest in SLE patients; second highest in unaffected sisters of SLE patients; third highest in healthy African-American women and lowest in healthy Caucasian women and malaria-exposed West African women.

Conclusion: This risk index may be useful in early interventions to prevent SLE. In addition, it suggests new therapeutic approaches for the treatment of SLE.
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http://dx.doi.org/10.1186/s10020-018-0019-4DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6016868PMC
June 2018

Analysis of menstrual effluent: diagnostic potential for endometriosis.

Authors:
Laura A Warren Andrew Shih Susana Marquez Renteira Tamer Seckin Brandon Blau Kim Simpfendorfer Annette Lee Christine N Metz Peter K Gregersen

Mol Med 2018 03 19;24(1). Epub 2018 Mar 19.

The Donald and Barbara Zucker School of Medicine at Hofstra/Northwell, 500 Hofstra Blvd, Hempstead, NY, 11549, USA.

Background: Endometriosis is a chronic and underdiagnosed disease which affects 5-10% of women of childbearing age and is characterized by growth of endometrial tissue outside of the uterus, most often in the peritoneal cavity. Delay in diagnosis is a major problem for management of this disorder, and treatment is often not initiated until the disease has progressed for many years. Although the exact etiology of endometriosis remains unknown, retrograde menstruation is recognized as a common underlying factor leading to the deposit of menstrual effluent (ME) into the peritoneal cavity. Differences in the cellular biology and genetics of the cells within ME are therefore likely to explain why endometriosis develops in only a subset of women.

Methods: Patients with and without endometriosis were consented to provide ME. ME was analyzed by flow cytometry for CD45- and CD45+ cell populations or used to isolate stromal fibroblast cells. ME-derived stromal fibroblast cells were assessed using decidualization assays following the addition of cAMP and IGFBP-1 concentrations in the culture supernatants were measured by ELISA. In addition, RNA was collected and analyzed by RNA-Seq and qPCR for markers of decidualization and to identify differentially expressed genes in ME-derived stromal fibroblast cells obtained from controls and subjects with endometriosis (±cAMP).

Results: Flow cytometry analysis of cell subsets within the CD45+ fraction of ME revealed a significant decrease in the number of uterine NK cells in endometriosis patients compared with controls (p < 0.01). No other significant differences within either the CD45+ or CD45- cell populations were observed. Most strikingly, ME-derived stromal fibroblast cells cultured from endometriosis subjects showed impaired decidualization potential compared with controls. Highly significant differences in decidualization response were detected by measuring IGFBP-1 production at multiple time points after cAMP stimulation (p = 0.0025 at 6 h; p = 0.0045 at 24 h; p = 0.0125 at 48 h). RNA-Seq and qPCR analyses were used to identify genes differentially expressed by ME-derived stromal fibroblast cells obtained from endometriosis and control subjects.

Conclusions: Menstrual effluent can be useful for investigating the pathobiology of endometriosis and for developing a non-invasive diagnostic for endometriosis which may lead to earlier and more effective treatments for this common disorder.
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http://dx.doi.org/10.1186/s10020-018-0009-6DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6016873PMC
March 2018

Methods for high-dimensional analysis of cells dissociated from cryopreserved synovial tissue

Authors:
Laura T Donlin Deepak A Rao Kevin Wei Kamil Slowikowski Mandy J McGeachy Jason D Turner Nida Meednu Fumitaka Mizoguchi Maria Gutierrez-Arcelus David J Lieb Joshua Keegan Kaylin Muskat Joshua Hillman Cristina Rozo Edd Ricker Thomas M Eisenhaure Shuqiang Li Edward P Browne Adam Chicoine Danielle Sutherby Akiko Noma Chad Nusbaum Stephen Kelly Alessandra B Pernis Lionel B Ivashkiv Susan M Goodman William H Robinson Paul J Utz James A Lederer Ellen M Gravallese Brendan F Boyce Nir Hacohen Costantino Pitzalis Peter K Gregersen Gary S Firestein Soumya Raychaudhuri Larry W Moreland V Michael Holers Vivian P Bykerk Andrew Filer David L Boyle Michael B Brenner Jennifer H Anolik

Arthritis Res Ther 2018 07 11;20(1):139. Epub 2018 Jul 11.

University of Rochester Medical Center, Rochester, NY, 14642, USA.

Background: Detailed molecular analyses of cells from rheumatoid arthritis (RA) synovium hold promise in identifying cellular phenotypes that drive tissue pathology and joint damage. The Accelerating Medicines Partnership RA/SLE Network aims to deconstruct autoimmune pathology by examining cells within target tissues through multiple high-dimensional assays. Robust standardized protocols need to be developed before cellular phenotypes at a single cell level can be effectively compared across patient samples.

Methods: Multiple clinical sites collected cryopreserved synovial tissue fragments from arthroplasty and synovial biopsy in a 10% DMSO solution. Mechanical and enzymatic dissociation parameters were optimized for viable cell extraction and surface protein preservation for cell sorting and mass cytometry, as well as for reproducibility in RNA sequencing (RNA-seq). Cryopreserved synovial samples were collectively analyzed at a central processing site by a custom-designed and validated 35-marker mass cytometry panel. In parallel, each sample was flow sorted into fibroblast, T-cell, B-cell, and macrophage suspensions for bulk population RNA-seq and plate-based single-cell CEL-Seq2 RNA-seq.

Results: Upon dissociation, cryopreserved synovial tissue fragments yielded a high frequency of viable cells, comparable to samples undergoing immediate processing. Optimization of synovial tissue dissociation across six clinical collection sites with ~ 30 arthroplasty and ~ 20 biopsy samples yielded a consensus digestion protocol using 100 μg/ml of Liberase™ TL enzyme preparation. This protocol yielded immune and stromal cell lineages with preserved surface markers and minimized variability across replicate RNA-seq transcriptomes. Mass cytometry analysis of cells from cryopreserved synovium distinguished diverse fibroblast phenotypes, distinct populations of memory B cells and antibody-secreting cells, and multiple CD4 and CD8 T-cell activation states. Bulk RNA-seq of sorted cell populations demonstrated robust separation of synovial lymphocytes, fibroblasts, and macrophages. Single-cell RNA-seq produced transcriptomes of over 1000 genes/cell, including transcripts encoding characteristic lineage markers identified.

Conclusions: We have established a robust protocol to acquire viable cells from cryopreserved synovial tissue with intact transcriptomes and cell surface phenotypes. A centralized pipeline to generate multiple high-dimensional analyses of synovial tissue samples collected across a collaborative network was developed. Integrated analysis of such datasets from large patient cohorts may help define molecular heterogeneity within RA pathology and identify new therapeutic targets and biomarkers.
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http://dx.doi.org/10.1186/s13075-018-1631-yDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6042350PMC
July 2018

Lineage-Specific Functionality of an Interferon Regulatory Factor 5 Lupus Risk Haplotype: Lack of B Cell Intrinsic Effects.

Authors:
Justine Calise Susana Marquez Renteria Peter K Gregersen Betty Diamond

Front Immunol 2018 7;9:996. Epub 2018 May 7.

Laboratory of Autoimmune & Musculoskeletal Diseases, The Feinstein Institute for Medical Research, Center for Autoimmune, Musculoskeletal, and Hematopoietic Diseases, Northwell Health, Manhasset, NY, United States.

Interferon regulatory factor 5 (IRF5) is widely recognized as a risk locus for systemic lupus erythematosus (SLE). Risk gene and IRF5 activation is triggered through toll-like receptor signaling. In myeloid cells, this leads to production of type I interferon and inflammatory cytokines, with enhanced production in cells of individuals harboring IRF5 risk alleles. Mouse models have also demonstrated the importance of IRF5 in B cell function, particularly plasma cell differentiation and isotype switching. Here, we evaluated the major SLE risk haplotype of IRF5 on the functional attributes of freshly isolated B cells from human subjects who do not have evidence of SLE or other forms of autoimmunity. We took this approach to avoid the complications of studying genotype-phenotype relationships in B cells that have been chronically exposed to an inflammatory disease environment before isolation. We focused on B cell endophenotypes that included gene expression, antibody secretion, class switching, and apoptotic susceptibility. We performed IRF5 overexpression studies, genetic reporter assays and electro-mobility shift assays on B and myeloid cell lines. Somewhat surprisingly, the results of our analyses indicate that IRF5 risk genotypes do not have a B cell intrinsic effect on these B cell functions. By contrast, we confirmed that the IRF5 risk and non-risk haplotypes exert differential effects in myeloid cells, including an increased susceptibility to apoptosis conferred by the risk haplotype. We also demonstrated an increased binding of the transcription factor specificity protein 1 to an insertion/deletion present in the risk haplotype. Our findings raise the specter that genetic risk alleles can have complex and unexpected lineage-specific effects, and these must be carefully considered when guiding or developing therapies based on understanding disease risk haplotypes.
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http://dx.doi.org/10.3389/fimmu.2018.00996DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5949527PMC
July 2019

Persons with Chronic Spinal Cord Injury Have Decreased Natural Killer Cell and Increased Toll-Like Receptor/Inflammatory Gene Expression.

Authors:
Paige Herman Adam Stein Katie Gibbs Ilya Korsunsky Peter Gregersen Ona Bloom

J Neurotrauma 2018 08 5;35(15):1819-1829. Epub 2018 Apr 5.

1 The Feinstein Institute for Medical Research , Northwell Health.

Infections are the leading cause of death for individuals with traumatic spinal cord injury (SCI). Along with increased infection rates, inflammation is often also observed in persons with chronic SCI. Together, immunological changes post-SCI are also poised to impede neurological recovery and mediate common medical consequences of SCI, including atherogenesis and neuropathic pain. The molecular mechanisms contributing to increased infection susceptibility and inflammation in persons living with SCI are poorly understood. Here, we used tools of functional genomics to perform a pilot study to compare whole-blood gene expression in individuals with chronic SCI (≥1 year from initial injury; N = 31) and uninjured individuals (N = 26). We identified 1815 differentially expressed genes in all SCI participants and 2226 differentially expressed genes in persons with SCI rostral to thoracic level 5, compared to uninjured participants. This included marked downregulation of natural killer cell genes and upregulation of the proinflammatory Toll-like receptor signaling pathway. These data provide novel mechanistic insights into the causes underlying the symptoms of immune dysfunction in individuals living with SCI.
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http://dx.doi.org/10.1089/neu.2017.5519DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6033303PMC
August 2018

Gene-level association analysis of systemic sclerosis: A comparison of African-Americans and White populations.

Authors:
Olga Y Gorlova Yafang Li Ivan Gorlov Jun Ying Wei V Chen Shervin Assassi John D Reveille Frank C Arnett Xiaodong Zhou Lara Bossini-Castillo Elena Lopez-Isac Marialbert Acosta-Herrera Peter K Gregersen Annette T Lee Virginia D Steen Barri J Fessler Dinesh Khanna Elena Schiopu Richard M Silver Jerry A Molitor Daniel E Furst Suzanne Kafaja Robert W Simms Robert A Lafyatis Patricia Carreira Carmen Pilar Simeon Ivan Castellvi Emma Beltran Norberto Ortego Christopher I Amos Javier Martin Maureen D Mayes

PLoS One 2018 2;13(1):e0189498. Epub 2018 Jan 2.

Department of Internal Medicine, Division of Rheumatology, University of Texas McGovern Medical School, Houston, TX, United States of America.

Gene-level analysis of ImmunoChip or genome-wide association studies (GWAS) data has not been previously reported for systemic sclerosis (SSc, scleroderma). The objective of this study was to analyze genetic susceptibility loci in SSc at the gene level and to determine if the detected associations were shared in African-American and White populations, using data from ImmunoChip and GWAS genotyping studies. The White sample included 1833 cases and 3466 controls (956 cases and 2741 controls from the US and 877 cases and 725 controls from Spain) and the African American sample, 291 cases and 260 controls. In both Whites and African Americans, we performed a gene-level analysis that integrates association statistics in a gene possibly harboring multiple SNPs with weak effect on disease risk, using Versatile Gene-based Association Study (VEGAS) software. The SNP-level analysis was performed using PLINK v.1.07. We identified 4 novel candidate genes (STAT1, FCGR2C, NIPSNAP3B, and SCT) significantly associated and 4 genes (SERBP1, PINX1, TMEM175 and EXOC2) suggestively associated with SSc in the gene level analysis in White patients. As an exploratory analysis we compared the results on Whites with those from African Americans. Of previously established susceptibility genes identified in Whites, only TNFAIP3 was significant at the nominal level (p = 6.13x10-3) in African Americans in the gene-level analysis of the ImmunoChip data. Among the top suggestive novel genes identified in Whites based on the ImmunoChip data, FCGR2C and PINX1 were only nominally significant in African Americans (p = 0.016 and p = 0.028, respectively), while among the top novel genes identified in the gene-level analysis in African Americans, UNC5C (p = 5.57x10-4) and CLEC16A (p = 0.0463) were also nominally significant in Whites. We also present the gene-level analysis of SSc clinical and autoantibody phenotypes among Whites. Our findings need to be validated by independent studies, particularly due to the limited sample size of African Americans.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0189498PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5749683PMC
February 2018