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Background:Sjögren’s Syndrome (SjS) is a complex autoimmune disease predominantly affecting the salivary and lacrimal glands, leading to hallmark symptoms such as dry eyes and mouth. Its distinctive clinical manifestations, coupled with symptom overlap with other disorders, make its diagnosis challenging. Traditional diagnostic criteria, as outlined by the American College of Rheumatology (ACR), encompass clinical assessments, including Anti-Ro/SSA antibody levels, focus score, and quantification of dry eye and mouth. Despite their utility, these criteria often suffer from issues of reproducibility and sensitivity. This underscores the pressing need for more advanced and reliable diagnostic techniques in the field.Objectives:In this work we developed a deep learning algorithm that combines clinical data with histopathological images of minor salivary glands in order to predict for Sjögren’s syndrome. By analyzing H&E stained labial gland biopsies using a Convolutional Neural Network (CNN) encoder in conjunction with a Multi-Layer Perceptron (MLP) and the ACR criteria. This strategy aims to improve diagnostic precision above and beyond the capabilities of traditional techniques, providing a notable improvement in the diagnosis and treatment planning of Sjögren’s Syndrome patients.Methods:Data for this study was sourced from the DIApSS (Diagnostic Suspicion of Primitive Sjögren’s Syndrome - Brest Cohort, NCT03681964) observational study. Our dataset comprised 167 patients, 95 confirmed Sjögren’s patients and 72 non-Sjögren’s participants. Patients were split into three groups training: 99 for training, 33 for validation, and 33 for testing. Our pipeline includes a Multi-Layer Perceptron (MLP) and a Convolutional Neural Network (CNN) encoder, implemented using PyTorch. The MLP processes clinical data: gender, xerophthalmia symptoms, Schirmer’s test, and Anti-Ro/SSA (UA/mL) and the CNN encoder, which processes H&E stained labial gland biopsy images. The combined output of both models is then fed into a classification head for prediction.Results:Evaluating the diagnostic precision using the clinical data alongside H&E Whole Slide Images (WSI), we obtained an Area Under the Curve (AUC) of 0.98, Accuracy of 0.86, kappa score 0.69, and a recall of 0.86. Cohen’s kappa score of 0.69 indicates that the model’s predictions show a moderate degree of agreement that goes beyond chance.Conclusion:This study successfully demonstrates the potential of integrating clinical data and histopathological images to obtain significant diagnostic accuracy. This approach paves the way for more reliable and reproducible diagnoses of Sjögren’s Syndrome.In future studies we will focus on refining the trained models and exploring further the analysis of the histopathological images.REFERENCES:NILAcknowledgements:NIL.Disclosure of Interests:None declared.

Background:Primary Sjögren’s syndrome (pSS) is an autoimmune disease, known for its disabling effect and chronic course. One of the peculiar symptoms is the lachrymal glands dryness associated often but not limited to dry mouth, dental disorders, joint pain, fatigue, and, in severe cases, systemic complications. The most relevant clinical feature is the infiltration of lymphocytes within the salivary glands and the development of an autoimmune endocrinopathy that can overstimulate lymphocytes until the development of lymphoma in 5% of the patients. pSS treatments are limited, and a deeper disease understanding is mandatory. Recently, Soret et al1 proposed a novel classification of pSS patients, in line with the PRECISESADS project2, aiming to reclassify the autoimmune diseases based on their biology more than the clinical features.Objectives:The ‘interferon’ cluster 1 (C1), ‘healthy-like’ cluster 2 (C2), ‘lymphoid’ cluster 3 (C3) and ‘inflammatory’ cluster 4 (C4) are analysed with novel datasets and omics from the same patients of the Soret et al study, including RNA-seq data from B lymphocytes and metabolomics data from plasma and urine. The multi-omics data integration by the MOFA algorithm is applied to extract factors able to catch the common variance from the novel and older omics. This study aims to extend the previous work and identify metabolomics markers easily obtainable with routine analysis to classify new pSS patients and provide the best care.Methods:Bioinformatics analyses were performed on the PRECISESADS datasets, including transcriptomics, metabolomics, methylomics and clinical data from over 300 pSS patients. The B-cell transcriptome was analysed using DESeq and GSEA. Plasma and urine metabolomics peak changes were quantified, statistically tested, and annotated using the Ceu Mass Mediator database. Common sources of variation among all the databases were identified using the MOFA integration analysis for each cluster, and the factor tested to be significantly discriminant to CTRLs. The clustering was performed in B-cell, plasma and urine data by linear discriminant analysis (LDA).Results:The B cell transcriptome highlighted the clusters C1 and C3 as the most affected by the interferon pathway, while C2 and C4 showed few differences compared to CTRLs. The cluster C4, marked by lymphopenia, had a low contribution of B lymphocytes in driving this patient cluster. Glycosylation genes (GALNTL6, MGAT3 and ENOSF1) contributed to the C2 and C4 differences among the clusters, while C1 and C3 by interferon signalling. Metabolomics analysis shed light on differences only in the plasma C1 cluster, where Lysophosphatidylcholine (LysoPC), phosphatidylinositol (PI) and neutral sphingolipids were upregulated, together with metabolites related to protein and nucleotide degradation. All clusters had a MOFA factor linked to interferon except the C2, where a single significant factor driven by B cell genes was associated with epigenetic modifications. Cluster 4 showed a factor associated with apoptosis in line with the lymphopenia, and carnitine complex showed a protective role in C1, C3, and C4 clusters, always contributing against their phenotype. LDA unveiled the drivers of the cluster differences, including interferon for B lymphocytes and cholines-associated lipids and phosphatidylinositol for plasma.Conclusion:This study provided novel details about the clustering of pSS patients observed in other studies1,2. B lymphocytes in cluster C4 showed little difference compared to CTRLs, while glycosylation, interferon signalling and epigenetics are proposed as drivers in B cell alteration in the other Sjogren clusters. PI, choline lipids and carnitine were identified in plasma as discriminant markers in the pSS clustering prediction, making them promising for their easy clinical measurement.REFERENCES:[1] Soret, P. A new molecular classification to drive precision treatment strategies in primary Sjögren’s syndrome. Nat Commun 12, 3523 (2021).[2] Barturen, G. et al. Integrative Analysis Reveals a Molecular Stratification of Systemic Autoimmune Diseases. Arthritis Rheumatol. Hoboken NJ 73, 1073–1085 (2021).Acknowledgements:NIL.Disclosure of Interests:CRISTIAN IPERI: None declared, Alvaro Fernández-Ochoa: None declared, Jacques-Olivier Pers: None declared, Nathan Foulquier: None declared, Guillermo Barturen: None declared, Marta Alarcon-Riquelme As part of the public European project PRECISESADS from the Innovative Medicines Initiative Joint Undertaking under Grant Agreement Number 115565. Innovative Health initiative from the European Union with in-kind contributions from the pharmaceutical industry (Sanofi, Roche, GSK, BMS, Novartis, Janssen, Tekada, Astra Zeneca and Pfizer. Payments are within the project and only BMS has made direct payments to her institution for personnel., Divi Cornec: None declared, Anne Bordron: None declared, Christophe Jamin: None declared.

BackgroundPrimary Sjögren’s syndrome (pSS) is a chronic autoimmune disease often diagnosed years after appearance of the first symptoms. Therefore, little is known about initial development of the disease. Anti-Ro antibodies can be found years before the first symptoms appear.ObjectivesTo identify individuals at risk for pSS and follow up over 10 years to examine symptoms and immunopathology before and during the development of pSS.MethodsThis ongoing longitudinal study enrolled individuals at risk for developing pSS but have not met the ACR-EULAR classification criteria. We recruited three groups: A.) Anti-Ro positive individuals without any symptoms for an underlying systemic autoimmune disease; B.) First degree relatives of patients with a diagnosis of pSS AND an abnormal immunologic laboratory value (ANA ≥ 1:160 and/or anti-Ro+ and/or rheumatoid factor+) C.) Individuals with at least one feature of the ACR-EULAR classification criteria for pSS, but not fulfilling the criteria.At screening and at annual follow-up visits, demographic data, blood, urine and saliva samples were obtained. Salivary flow, Schirmer’s test, and salivary gland ultrasonography (SGUS) using Hocevar-Score were performed. A lip salivary gland biopsy (LSB) was obtained at the screening visit and repeated once when typical symptoms appeared within the follow-up visits. The primary endpoint was the development of definite pSS according to the ACR-EULAR classification criteria or a diagnosis of another systemic autoimmune disease.ResultsAfter two years, 53 subjects were recruited from all three groups, of whom 31 were enrolled in the study (Anti-SSA+ n=25, relatives n=4, incomplete n=2). Most of the 22 screening failures were relatives who did not have an abnormal immunologic laboratory value.28 individuals were female, they had a mean age of 49.6 ± 12.7 years (SD; standard deviation) and a mean body weight of 71.4 ± 15 kg. 11 individuals reported having a relative with an underlying autoimmune disease. The median ESSDAI was 0 (0-5; min-max). individuals had a pathological stimulated whole saliva flow (mean 3.7 ± 1.4 g) and eight had pathological Schirmer’s test (left eye: mean 13.6 ± 13.1 mm; right eye: 13.2 ±12.2 mm). Three of 14 individuals had a pathological SGUS score. All three individuals were recruited in the Anti-Ro+ group. At screening visit, 11 LSB were performed (mean focus score of 0.53 ± 1.2) and one individual reached a focus score ≥ 1.So far, 18 individuals have completed the two-year visit. Two of them had pathological stimulated whole saliva flow (mean 3.85 ± 1.4 g) and five of them a pathological Schirmer’s test (left eye: mean 15.4 ± 13 mm; right eye: 14.5 ± 13). The median ESSDAI was 0 [0-4]. Eight SGUS were performed with a mean Hocevar-Score of 5.6 ± 8.Within two years, four patients (13%) were diagnosed with pSS, one (3%) with systemic lupus erythematosus and one (3%) with rheumatoid arthritis.ConclusionThis longitudinal study for the first time provides us with data on the evolution of pSS. Future recruitment at multiple centers and longer follow-ups will shed light on the earliest phase of pSS.REFERENCES:NIL.Acknowledgements:NIL.Disclosure of InterestsNone Declared.

Background:The diagnosis of primary Sjögren’s syndrome (pSS) is established at a time when symptoms are already present due to tissue damage that has already occurred. Thus, most of our knowledge about the disease is based on a presumed late stage of the autoimmune process that is probably initiated years before the onset of symptoms [1]. This assumption is supported by the lymphopenia often seen in pSS, resulting from a reduced naive T cell compartment. A finding that is likely to be the result of prolonged, continuous immune activation [2].Objectives:To investigate the clinical, immunological and histological characteristics of individuals at risk of developing pSS.Methods:This ongoing, longitudinal study included 41 individuals who did not fulfil the ACR/EULAR classification criteria but were considered at risk of developing pSS (defined as anti-Ro antibody positive, OR autoantibody positive first-degree relatives of pSS patients OR patients meeting more than one point of the ACR/EULAR classification criteria). Participants underwent annual study visits for patient- reported outcome measures, pSS- specific clinical assessments and ultrasonography. In addition, laboratory assessments including immune phenotyping of peripheral blood by flow cytometry (FACS) were performed. The FACS measurements were compared with age- and gender-matched healthy controls (HC). Some subjects also underwent salivary gland biopsy followed by an immunohistological staining procedure. The primary endpoint of the study is the diagnosis of pSS or another systemic autoimmune disease (AID).Results:Results from 41 at- risk individuals (female ratio: 0.85; mean ± standard deviation, age: 51 ± 13.27) showed that sicca symptoms were reported by nearly three quarters of participants at baseline (median ± interquartile range, ESSPRI dryness domain: 2 ± 5). 4 individuals already reached the primary endpoint by being diagnosed with pSS (n=3) or rheumatoid arthritis (n=1), respectively. At baseline, only one individual showed pSS- specific changes in the salivary gland ultrasound defined by a Hocevar score above 17 points. When compared to HC, the FACS analysis showed a significant decrease of the CD4+ T cell pool, mostly affecting the naive compartment. This decrease was less profound than in patients with established pSS. The histological analysis already revealed typical features of pSS in some of the individuals at risk (pre- lymphoepithelial lesions in 2/7, defined by presence of intraepithelial B- lymphocytes; plasma cell shift in 3/7, defined by equal amounts of IgG- and IgA- expressing plasma cells).Conclusion:In this study, we show that individuals at risk of developing pSS already have clinical, immunological and histological features of the disease. The reduction of the naive CD4+ T cell compartment and subtle inflammation of the salivary glands may be a sign of a subclinical disease process that begins before the full clinical picture of pSS is established.REFERENCES:[1] Olsen NJ, Okuda DT, Holers VM, Karp DR. Editorial: Understanding the concept of pre-clinical autoimmunity. Frontiers in immunology. 2022;13:983310.[2] Fessler J, Fasching P, Raicht A, Hammerl S, Weber J, Lackner A, et al. Lymphopenia in primary Sjögren’s syndrome is associated with premature aging of naïve CD4+ T cells. Rheumatology (Oxford). 2021;60(2):588-97.Figure 1.Significant reduction of naive CD4+ T cells in patients at risk (PreSS) compared to age- and gender- matched healthy controls (HC). Patients with primary Sjögren’s syndrome (pSS) showed an even more marked reduction in the naive compartment at the time of diagnosis.Acknowledgements:NIL.Disclosure of Interests:None declared.

Background:The epidemiological and immunopathogenic association between Sjögren disease (SD) and non-Hodgkin’s lymphoma (NHL) is well-established. However, the epidemiological data on this complication mostly relies on data from tertiary centers, introducing selection bias and limiting comprehensive case capture.Objectives:This study aimed to evaluate accurately the characteristics of lymphoma in primary or associated SD using data from the French nationwide healthcare information system (“système national des données de santé” [SNDS]).Methods:SD patients were identified in the SNDS database based on a validated algorithm [1] (ICD10 code for SD and reimbursement for at least 2 SD-prescribed drugs). Primary SD was defined in the absence of an ICD10 code for other autoimmune diseases, and associated SD was defined otherwise. The study analyzed, primary, associated and all SD patients and lymphoma occurrences between 2009 and 2022. Analysis of risk factors of lymphoma included demographic variables such as age, sex, disease duration, and proxy reflecting disease activity including notably cryoglobulinemic vasculitis and monoclonal gammopathy of unknown significance (MGUS).Results:Patients with primary SD In the 30,221 patients with primary SD (65.7% of the total cohort of patients with SD; 88.8% women, median age 60 [48;70], median follow-up 11.6 years [9.4;13.2]; hydroxychloroquine use in 35.1%, methotrexate [11.5%], leflunomide [1.2%], azathioprine [4.4%], mycophenolate [3.1%], cyclophosphamide [0.4%], and rituximab [5.5%]), lymphoma occurred in 1228 patients with a prevalence of 4.1%. Marginal zone lymphomas (MZL) were the predominant lymphoma subtype (53.8%) (especially MALT-MZL [30.9%]) followed by diffuse large B-cell lymphoma (DLBCL) (25.4%), follicular lymphoma (FL) (12.1%), Hodgkin’s lymphoma (8.2%), and T/NK lymphoma (7.2%). Mortality was higher in patients with lymphoma (379/1228 [30.9%] versus 4335/28993 [15%], p<0.001). Preliminary risk factor analysis revealed associations with MGUS and cryoglobulinemic vasculitis: MGUS was observed in 3.2% of SD patients without lymphoma, compared to 10.8% in those with lymphoma (p < 0.001), and cryoglobulinemic vasculitis occurred in 2.2% of SD patients without lymphoma, as compared with 10% of patients with lymphoma (p < 0.001).Patients with associated SD In the 15,791 patients with associated SD (34.3% of the total cohort, including rheumatoid arthritis [20.4%], systemic lupus [10%], systemic sclerosis [6.2%], other connective tissue disorders [3.2%], and juvenile arthritis [0.3%]), 483 patients developed lymphoma (3.1%).Total SD cohort In the total SD cohort, including 46,012 patients with either primary or associated SD (89.4% women, median age 59 [47;69], median follow-up 11.8 years [9.5;13.2]), lymphoma occurred in 1749 patients, with a 3.8% prevalence. Lymphoma subtypes were distributed similarly to the primary SD cohort, and associations between lymphoma and MGUS and cryoglobulinemic vasculitis were observed. Mortality was higher in patients with lymphoma compared to those without lymphoma (30.9% versus 14.8% [p < 0.001]).Conclusion:This extensive epidemiological nationwide study demonstrates a 4% prevalence of lymphoma in primary SD, and 3% prevalence in associated SD. Preliminary risk factor analysis confirms the association between SD-related lymphoma and MGUS and cryoglobulinemic vasculitis.REFERENCES:[1] Seror et al. (2024 in press) Development of an Algorithm to Identify Sjögren’s Syndrome Patients in the French National Healthcare Claims Database. RMD Open.Acknowledgements:NIL.Disclosure of Interests:None declared.

BackgroundPolymyalgia rheumatica (PMR) is an inflammatory disease. But it’s pathophysiology and the impact of the treatments on immune cells are poorly known.ObjectivesOur objectives were to describe the immune cells of patients with PMR dependent from glucocorticoids and to analyze their evolution under tocilizumab.MethodsThe SEMAPHORE trial (NCT02908217, (1)) was a randomized control trial including patients with PMR dependent from glucocorticoids. Between inclusion and week 12, patients were blindly treated with either tocilizumab infusion or placebo infusion every 4 weeks associated to a tapering of glucocorticoids. Age and sex-matched healthy controls (HC) were recruited in the rheumatology department of the Brest University Hospital. HC did not have any history or presence of cancer, auto-immune disease or active infection and did not received treatment with a known impact on the immune system. We analyzed immune cells on whole fresh blood after red cell lysis on flow cytometry (Navios, Cytoflex, Beckmann Coulter) after staining for CD16, CD56, CD19, CD14, CD4, CD8, CD3, CD45, IgD, IgM, CD21, CD27, CD24, CD38, CD5 CD126, CD62L, CD45RA, CD127, CD25, in four different panels.ResultsSamples were obtained for 40 PMR patients and 34 HC. At inclusion, in PMR patients, compared to HC, CD14+CD16- classical monocytes were increased (82±1% vs 77±2%, p=0.01), CD14-CD16+ non-classical monocytes were decreased (4±0.3% vs 7±0.6%, p<0.0001), granulocytes were increased (63±2% vs 52±2%, p<0.0001), natural killer cells were decreased (8±1% vs 13±1%, p=0.007). B cells were decreased (8±0.6% vs 10±0.8%, p=0.03), but with an enrichment in CD27-IgD- senescent B cells (p<0.0001) and in CD21low B cells (p=0.04). T cells were increased (70±2% vs 64±2%, p=0.02) with an enrichment in CD4+ T cells (p=0.02) and in CD4+CD25highCD127- regulatory T cells (p<0.0001). At week 12, in PMR patients receiving tocilizumab therapy, compared to PMR patients receiving placebo, granulocytes were lower (58±5% vs 73±2%, p=0.006) and monocytes were higher (8±1% vs 5±0.5%, p=0.02).ConclusionIn patients with a PMR dependent from glucocorticoids, immune cells homeostasis is disturbed. Tocilizumab has an impact more pronounced on granulocytes and monocytes. Knowledge about immune disturbance in PMR might help to choose to use a targeted therapy when glucocorticoids are not sufficient.Reference[1]Devauchelle-Pensec V, Carvajal-Alegria G, Dernis E, Richez C, Truchetet ME, Wendling D, et al. Effect of Tocilizumab on Disease Activity in Patients With Active Polymyalgia Rheumatica Receiving Glucocorticoid Therapy: A Randomized Clinical Trial. JAMA. 20 sept 2022;328(11):1053‑62.Acknowledgements:NIL.Disclosure of InterestsGuillermo CARVAJAL ALEGRIA Speakers bureau: Abbvie, Lilly, Novartis, Chugai, Biogen, BMS, Galapagos, Pfizer, Consultant of: Abbvie, Lilly, Novartis, Galapagos, Pfizer, Sara Boukhlal: None declared, Sophie Hillion: None declared, Pierre Pochard: None declared, Emmanuelle Porchet: None declared, Alain Saraux Speakers bureau: Chugai, Consultant of: Chugai, Grant/research support from: Chugai, Sandrine Jousse-Joulin Speakers bureau: Chugai, Consultant of: Chugai, Grant/research support from: Chugai, THIERRY MARHADOUR: None declared, Dewi Guellec: None declared, Divi Cornec Speakers bureau: Chugai, Consultant of: Chugai, Grant/research support from: Chugai, Valerie Devauchelle-Pensec Speakers bureau: Chugai, Consultant of: Chugai, Grant/research support from: Chugai.

Background:Sjögren’s disease (SjD) presents an unmet medical challenge as there is currently no cure. Despite advances in understanding the immunopathogenesis of SjD, there is still a pressing need to identify novel biomarkers and therapeutic targets, for better patient stratification and personalized treatment.Objectives:To create a fully-detailed molecular interaction map (MIM) including all the signalling and molecular pathways implicated in SjD pathogenesis. To create a large-scale mechanistic model to enable in silico simulations of perturbations including drug interventions, and the generation of hypothesis-driven predictions.Methods:Differential expression analysis was performed on blood samples from SjD patients vs controls on 3 datasets: the publicly available GSE51092 and the accessible via the NECESSITY consortium UKPSSR and PRECISEADS datasets. GSE51092 contains transcriptomic data of 190 SjD patients and 32 controls, UKPSSR of 151 SjD patients and 29 controls, and PRECISESADS, RNASeq data for 304 SjD patients and 341 controls. Pathway enrichment analysis was subsequently performed using GSEA and the Reactome pathway database[1,2]. Additional literature-based evidence was used to develop a molecular interaction map, combining the results of the previous analytical steps. The SjD specific map was then converted into a Boolean model using the CaSQ tool[3]. Logic rules based on Boolean algebra are used to describe every interaction between the molecular entities.Results:Our analysis unveiled a set of differentially expressed genes (DEG) and related pathways associated with immune dysregulation and inflammatory responses in SjD. We obtained a total of 1625 DEG, 725 DEG from PRECISESADS, 1161 DEG from GSE51092 and 239 DEG from UKPSSR, with 25 DEG common for all three datasets. Nine common DEG were associated with Interferon signalling. Twenty-one pathways were identified with both GSEA and Reactome-based analysis. The building of the SjD MIM was performed based on literature search and the 21 identified pathways, from the data analysis. The MIM includes so far 16 pathways (5 present in the identified pathways and 11 literature-mined), comprising 187 species (genes, RNAs proteins) and 132 reactions. The SjD-specific Boolean model obtained after conversion of the SjD MIM, represents a more compact and fully executable version of the SjD molecular network, containing 111 nodes and 130 edges. In Sjögren-specific conditions the model is able to reproduce the activation of main inflammation pathways and predict the inflammatory response.Conclusion:We have built the first SjD-specific MIM integrating omic data analyses and information from literature-based evidence and pathway enrichment analysis. The current SjD map contains hallmark disease pathways and is constantly being enriched with data-driven highlighted pathways. The preliminary computational model based on the SjD map is able to reproduce inflammatory response scenarios, however further training is needed to improve performance and robustness.REFERENCES:[1] Fabregat, A. et al. The Reactome pathway Knowledgebase. Nucleic Acids Research 44, D481–D487 (2016).[2] Subramanian, A. et al. Gene set enrichment analysis: A knowledge-based approach for interpreting genome-wide expression profiles. Proceedings of the National Academy of Sciences 102, 15545–15550 (2005).[3] Aghamiri, S. S. et al. Automated inference of Boolean models from molecular interaction maps using CaSQ. Bioinformatics 36, 4473–4482 (2020).Figure 1.Data analysis revealed 1625 DEG from three different datasets of expression data in Sjögren patients. Pathway enrichment analysis using GSEA and Reactome database identified 21 common enriched pathways. These pathways along with literature-based search were used as the starting point for the building of the Sjögren molecular interaction map (MIM). The first version of the map consists of 187 biomolecules organized in 16 pathways (5 data-driven and 11 literature -mined) and has served as a template for the inference of a discrete computational model. The model comprises 111 biomolecules and 130 interactions.Acknowledgements:I express my gratitude to the NECESSITY consortium members for the materials they made available to me, which enabled me to carry out this research. I would also like to thank the doctoral school SDSV of Paris-Saclay University and Genopole for financing respectively my PhD studies and formations I followed.Disclosure of Interests:Sacha E Silva-Saffar: None declared, Jacques-Eric Gottenberg BMS, Pfizer, Lilly, Abbvie, AstraZeneca, Gilead, Galapagos, MSD, Roche-Chugai, Sanofi, UCB, Michele Bombardieri: None declared, Divi Cornec: None declared, Jacques-Olivier Pers: None declared, Marta Alarcon-Riquelme: None declared, Philippe MOINGEON Sanofi, Stallergenes, Servier, Michael Barnes: None declared, Sandra Ng: None declared, Wan-Fai Ng GlaxoSmithKline, MedImmune, Novartis and BMS, Abbvie, Resolves Therapeutics, Nascient, Xavier Mariette Sanofi, Servier, BMS, Gaetane Nocturne Amgen, Novartis, Anna Niarakis Sanofi.

Background:The SEMAPHORE trial[1] was a randomized controlled prospective study to assess the safety and efficacy (success defined by PMR-AS≤10 and GC≤5mg or GC decrease ≥10mg/day) of intra venous (i.v.) tocilizumab (TCZ) in glucocorticoids (GC)-dependent polymyalgia rheumatica (PMR). Tocilizumab was shown to reduce GC while improving clinical and biological inflammatory parameters at 24 weeks. After the 24-week study, patients entered a double-blind extension.Objectives:In this study, we aimed to assess after the 24th week the relapse rate and markers of patients who received a 6-month TCZ treatment and achieved remission.Methods:Among 101 patients randomly 1:1 allocated to receive i.v. 8mg/kg TCZ or placebo for 24 weeks in the SEMAPHORE trial, 49 received TCZ and 33 of them succeed (Figure 1). All 33 patients but one stopped TCZ at week 24 and they visited at week 32, with optional follow-up visits every 8 weeks until week 48. The relapse was defined by the failure of the primary composite outcome during follow-up or the need for ≥1 TCZ infusion. Results are presented by proportion of patients achieving success at each visit and the outcome over time using a Kaplan Meier curve.Results:Among the 33 TCZ group patients in remission at week 24, 7 stopped follow-up before the 48th week, 5 of the other 26 subjects (19.2%) sustained remission in the 6 following months, 21 (80.8%) relapsed. Median time to relapse was 15 weeks (interquartile range: 8-25). 15 patients were considered in relapse because of PMR-AS≥10, 4 for isolated CRP elevation, 2 after the clinician’s decision. Among the 16 patients treated with TCZ but not in remission at 24th week, 4 of them (25%) achieved the primary outcome after continuing TCZ for 24 other weeks.Conclusion:Among patients with GC-dependent PMR, in remission after a 6-month TCZ treatment, only a quarter remained relapse-free after treatment discontinuation. This study suggests that a 6-month treatment is not enough to withdraw TCZ. Further studies assessing the required duration of anti-IL6-receptor treatment to limit PMR relapses are needed.REFERENCES:[1] Devauchelle-Pensec, V. et al. Effect of tocilizumab on Disease Activity in patients with active polymyalgia rheumatica receiving glucocorticoid therapy: A Randomized Clinical Trial. JAMA 328, 1053–1062 (2022).Figure 1.Flow chart of the patients included in the SEMAPHORE trial, orientation of patients after the 24th weekFigure 2.Relapse-free survival among GC-dependant PMR patients in remission after a 6-month treatment of TocilizumabFootnotes: Remission was defined by PMR-AS<10, and GC≤5mg/j or daily GC dose decreased by ≥10mg. Day 0 was the visit at 24th week in the SEMAPHORE trialAbbreviations: PMR: Polymyalgia rheumatica; PMR-AS: Polymyalgia rheumatica activity scoreAcknowledgements:We thank the French rheumatologists, particularly those from the French University Hospital VICTOR HUGO (InnoVation en reCherche OsTeo-aRticulaire des Hôpitaux Universitaires du Grand Ouest) network (srouest.fr), and the general practitioners who referred their patients to this trial. We are grateful to the Clinical Investigations Center (CIC) 1412, Institut National de la Santé et de la Recherche Médicale (INSERM), Brest, France, for centralizing the trial data and to Audrey le Goff, MS; Adrien Clarysse, MS; and Valentine Guiton, MS, of the Brest University Hospital research board (DRCI) at the Brest University Hospital, who received no compensation for their role in the study.Disclosure of Interests:Baptiste Chevet Galapagos, I wrote congress review for a journal they own, I received support outside of this work from Amgen, Abbviie, Novartis, Santi, Sandoz, Souki Aghiles: None declared, Nowak Emmanuel Dr Nowak reported receiving nonfinancial support from Roche-Chugai Pharmaceutical, which donated the infusion form of tocilizumab during the conduct of the study., Dr Nowak reported receiving grants from the French National Program for Clinical Research, Guillermo Carvajal Alegria Dr Carvajal Alegria reported receiving personal fees from Chugai Pharmaceutical outside the submitted work., Emmanuelle Dernis Dr Dernis reported receiving personal fees from Novartis, Bristol Myers Squibb, UCB, AbbVie, Nordic Pharma, Amgen, and Janssen outside the submitted work., Christophe Richez Dr Richez reported receiving personal fees from Sanofi, personal fees from Lilly, and personal fees from Novartis outside the submitted work., Marie-Elise Truchetet: None declared, Daniel Wendling Dr Wendling reported personal fees from AbbVie, Bristol Myers Squibb, MSD, Pfizer, Chugai Pharmaceutical, Amgen, Nordic Pharma, UCB, Novartis, Janssen, Eli Lilly, Grunenthal, and Galapagos outside the submitted work, Eric Toussirot: None declared, Aleth Perdriger: None declared, Jacques-Eric Gottenberg Dr Gottenberg reported receiving personal fees from Roche-Chugai Pharmaceutical, Sanofi, Pfizer, Eli Lilly, Gilead, and Abbvie and grants from Bristol Myers Squibb outside the submitted work., Renaud Felten: None declared, Bruno Fautrel Dr Fautrel reported receiving personal fees from Roche Chugai Pharmaceutical during the conduct of the study., Anne Lohse: None declared, Laurent Chiche: None declared, Pascal Hilliquin: None declared, Catherine Le Henaff Bourhis: None declared, Dervieux Benjamin: None declared, Guillaume Direz Dr Direz reported receiving personal fees from Novartis and personal fees from Roche-Chugai Pharmaceutical outside the submitted work., Isabelle Chary-Valckenaere: None declared, Divi Cornec: None declared, Dewi Guellec: None declared, Thierry Marhadour: None declared, Alain Saraux Dr Saraux reported receiving grants from Roche-Chugai Pharmaceutical and grants from the French National Program for Clinical Research during the conduct of the study and personal fees from Nordic Galapagos, AbbVie, Eli Lilly, Nordic, and Roche-Chugai and grants from Eli Lilly outside the submitted work., Valerie Devauchelle-Pensec Dr Devauchelle reported receiving personal fees from Chugai Pharmaceutical and AbbVie a, Dr Devauchelle-Pensec reported grants and personal fees from Novartis during the conduct of the study and personal fees from Galapagos, Pfizer, Bristol Myers Squibb, Janssen, and AbbVie outside the submitted work.

BackgroundGiant cell arteritis (GCA) is an inflammatory disease marked by granulomatous inflammation of the large- and medium-vessels. GCA exclusively affects older patients above 50 years old, suggesting a role of senescence in the pathobiology of GCA. IL-6 receptor blocking therapy has been shown to be effective treatment for GCA.(1) More recently, a phase two randomized control trial has shown the efficacy of GM-CSF receptor blocking therapy for the treatment of GCA.(2) Both IL-6 and GM-CSF are senescence-associated secretory phenotype (SASP) cytokines,(3) implicating the roles of cellular senescence and SASP in the immunopathology of GCA.ObjectivesWe seek to find clues of senescent and SASP in the inflamed arteries of GCA patients.MethodsImmunohistochemical stainings of the senescence markers p21 WAF1/CIP1 and p16/INK4A as well as SAPS cytokines IL-6 and GM-CSF were performed on temporal artery biopsies (TABs; n=8) from patients with giant cell arteritis. Non-inflamed arteries from age-matched controls were included (n=14). Stainings were scored quantitatively by using QuPath software. Opal based multiplex fluorescent stainings were performed to confirm the colocalization of p21 WAF1/CIP1 and p16/INK4A with IL-6 and GM-CSF.ResultsAll four markers were detected in the inflamed GCA temporal arteries, most notably by giant cells surrounding the intimal lamina elastica although expression by other cells was also detected throughout the vessels. The expression of all four markers were significantly lower in control arteries. Cells positive for p21 WAF1/CIP1 and p16/INK4A that co-express IL-6 and GM-CSF were detected throughout the vessels.ConclusionOur data suggest the presence of activated senescence pathways at the site of vascular inflammation in GCA and support further research into the role of senescence in GCA.References[1]Maz M, Chung SA, Abril A, Langford CA, Gorelik M, Guyatt G, Archer AM, Conn DL, Full KA, Grayson PC, et al. 2021 American College of Rheumatology/Vasculitis Foundation Guideline for the Management of Giant Cell Arteritis and Takayasu Arteritis. Arthritis Rheumatol 2021; 73:1349–1365.[2]Cid MC, Unizony SH, Blockmans D, Brouwer E, Dagna L, Dasgupta B, Hellmich B, Molloy E, Salvarani C, Trapnell BC, et al. Efficacy and safety of mavrilimumab in giant cell arteritis: a phase 2, randomised, double-blind, placebo-controlled trial. Ann Rheum Dis 2022; 81:653–661.[3]Coppé JP, Desprez PY, Krtolica A, Campisi J. The Senescence-Associated Secretory Phenotype: The Dark Side of Tumor Suppression. Annu Rev Pathol 2010; 5:99.AcknowledgementsThe two (or more) of the/several author(s) of this publication is/are (a) member(s) of the European Reference Network Rare Immunodeficiency, Autoinflammatory and Autoimmune Diseases Network (ERN RITA) and the European Reference Network Rare Connective Tissue and Musculoskeletal Diseases Network (ERN ReCONNECT).Disclosure of InterestsWilliam Febry Jiemy: None declared, Yannick van Sleen: None declared, Jacoba Carolien Graver: None declared, Sarah A. Pringle: None declared, Elisabeth Brouwer Speakers bureau: EB reports personal fees from Roche, outside the submitted work., Consultant of: EB reports personal fees from Roche, outside the submitted work., Kornelis van der Geest Speakers bureau: KSMvdG reports personal fees from Roche, outside the submitted work., Consultant of: KSMvdG reports personal fees from Roche, outside the submitted work., Divi Cornec: None declared, Annemieke Boots: None declared, Maria Sandovici: None declared.

BackgroundAutoreactive proteinase 3 (PR3+) B cells have recently been phenotypically and functionally characterized, and the presence of defective central antigen-independent and peripheral antigen-dependent checkpoints in patients with ANCA-associated vasculitis (AAV) has been shown. This work aimed to investigate the central tolerance-checkpoint controlling immature PR3+ B cells in the bone marrow (BM), before their migration into the periphery as transitional B cells.ObjectivesWe investigated the presence and the specific phenotypic features of PR3+ B cells in BM mononuclear cells (BMMC) of non-vasculitis controls (No-AAV), comparing them to paired peripheral blood mononuclear cells (PBMC) of No-AAV and PBMC of PR3-AAV patients, and the central tolerance-checkpoint for PR3+ B cells.MethodsWe used a customized flow-cytometry assay, using PR3 as ligand to target autoreactive PR3+ B cells (PR3+B cells). Adult PR3-ANCA positive AAV (PR3-AAV) patients with a clinical diagnosis of granulomatosis with polyangiitis (GPA) or microscopic polyangiitis (MPA) were selected among consecutive subjects with AAV seen in our Rheumatology Unit. Subjects without vasculitis (No AAV) were selected among consecutive subjects undergoing bone marrow aspirate to exclude hematologic conditions of myeloid origin and eventually resulted healthy or in long-term complete remission during follow-up of myeloid neoplasms. PMBC from AAV patients and paired samples of BMMC and PBMC from No AAV were collected and analyzed.ResultsThe proportion of PR3+ B cells within BMMC (median [IQR25-75%]; 1.98%[1.77-2.75]) was higher than within PBMC of No-AAV (0.9%[0.63-1.44], p<0.01 by paired comparison) and similar to their proportion within PBMC of PR3-AAV patients (1.82%[1.66-3.21]; p>0.05). When focusing on immature/transitional CD24++CD38++B cells only in No-AAV, we observed distinct phenotypes within BMMC versus PBMC (i.e. higher proportion of CD27-CD10+ and lower expression of CD21, IgD, IgM within BMMC versus PBMC), representing two separate developmental steps of B cell maturation. Within CD24++CD38++ B cells, BMMC contained the greatest proportion of PR3+ B cells as compared to PBMC (3.35%[1.99-4.92] versus 1.23%[0.62-1.55], p<0.01). We observed a significant decline of the PR3+ fraction from T1-like/immature subset (IgD-IgM+; 2.80%[1.23-4.02]) to T2-like/early transitional subset (IgD+IgM+; 1.76%[0.96-2.68], p<0.01) in BMMC, while no significant reduction was observed between the latter subset and the transitional compartment of PBMC (1.26%[0.62-1.56], p>0.05).ConclusionTo prevent PR3-related autoimmunity, autoreactive PR3+ B cells pass a stringent selection in the BM, and their removal by central tolerance-checkpoint activity occurs mainly between T1-like/immature to T2-like/early transitional B cells of BMMC.References[1]Cornec D. Identification and phenotyping of circulating autoreactive proteinase 3-specific B cells in patients with PR3-ANCA associated vasculitis and healthy controls. J of Autoimmunity, 2017.[2]Berti A. Circulating autoreactive proteinase 3+ B cells and tolerance checkpoints in ANCA-associated vasculitis. JCI Insight, 2021.Acknowledgements:NIL.Disclosure of InterestsAlvise Berti Speakers bureau: GSK, Michele Tomasi: None declared, Isabella Pesce: None declared, Enrico Lista: None declared, Anna Guella: None declared, Giuseppe Paolazzi: None declared, Roberto Bortolotti: None declared, Guido Grandi: None declared, Sophie Hillion: None declared, Ulrich Specks: None declared, Divi Cornec: None declared.