17th Pediatric Rheumatology European Society Congress September 9-12, 2010 València, Spain Abstracts Page no. Oral Abstracts (1 – 36) xxx Poster Abstracts (1 – 306) xxx Clinical and Experimental Rheumatology 2011; xx: xxx-xxx. Oral abstracts 17th Pediatric Rheumatology European Society Congress Oral Abstracts RESULTS: This study had >80% to detect an odds ratio >1.25 for SNPs with allele frequencies >0.1. Two SNPs in the MVK gene, rs1183616 (ptrend=0.006 OR 1.17 95% CI 1.04-1.30) and rs7957619 (ptrend=0.005 OR 1.23 95% CI 1.07- O 01 1.43) are significantly associated with JIA. These two SNPs are in modest linkage Distinctive gene expression in patients with juvenile spondylo- disequilibrium (r2=0.36, D’=1). Logistic regression of the two SNPs, after condi- arthropathy is related to autoinflammatory diseases tioning on the most significant SNP, found that the rs1183616 SNP was no longer significant (p=0.3), suggesting that the association is a single effect driven by the Marina Frleta, Lovro Lamot, Fran Borovecki, Lana Tambic Bukovac, Miroslav rs7957619 SNP. This SNP lies within exon 3 of the MVK gene and is a Serine to Harjacek Asparagine substitution at position 52. There was no significant evidence of a dif- Children’s Hospital Srebrnjak, Srebrnjak, Zagreb, Croatia ference in allele frequencies between the seven ILAR subtypes for the rs7957619 SNP (p=0.32). INTRODUCTION: Juvenile Spondyloarthropathies (jSpA) are characterized by One SNP at the 3’ end of the TNFRSF1A gene, which actually lies within the dysregulation of the inflammatory processes and bone metabolism which may be adjacent gene SLCNN1A, rs2228576, was associated with protection from JIA clarified by gene expression profiles. (ptrend=0.009 OR 0.87 95% CI 0.78-0.97). There was no significant evidence of a OBJECTIVES: To identify genes with disease-specific expression patterns of pa- difference in allele frequencies between the seven ILAR subtypes (p=0.94). None tients diagnosed with jSpA and healthy controls using microarray-based methods. of the 16 SNPs studied across the NALP1 gene were found to be associated with METHODOLOGY: Peripheral blood samples of 6 HLA-B27/B7, double posi- JIA susceptibility. tive“ patients (OR=14.9) with new onset, untreated disease were analyzed for ex- DISCUSSION: We have utilised the largest cohort of JIA cases available in Eu- pression patterns that correlated with disease characteristics using Human Genome rope and identified associations between JIA and polymorphisms 3’ of TNFRSF1 U133 PLUS 2.0 GeneChip, Affymetrix, (6x106 SNP’s). For comparison, gene ex- and within MVK. Replication of these findings in other JIA populations is required. pression profiles were obtained from 4 healthy controls. Real-time PCR was used These observations support the value of extrapolating from monogenic to complex for confirmation of gene expression differences. disease phenotypes. RESULTS: Statistical analysis of gene expression patterns identified 369 differ- entially expressed genes at statistical cutoffs fold change 1.5 (p<0.05, max>100). There were also 163 mRNAs with significantly increased expression, and 197 mR- NAs with significantly decreased expression. The genes represented by these probe O 03 sets were enriched for functions related to inflammatory modulation, MAP kinase pathway, TGF-beta family, as well as other enzymes and receptors (myosin light Clinical and microarray follow-up of SOJIA patients treated with chain kinase, NRLP3 (inflammasome), thrombomodulin, protein-tyrosin phospa- anakinra: lessons learned over the past 7 years hase, receptor type 2 (PTPRN2), TRAF1, and ZAP-70. Using network, DAVID, and GSEA analysis we discovered gene hubs among the differentially expressed Lynn Punaro, Alisa C. Gotte, Derek N. Blankenship, Matthew L. Stoll, Florence genes based on correlation of expression (T-cell regulation, energy metabolism, Allantaz, Zhaohui Xu, Virginia Pascual RNA processing). Texas Scottish Rite Hospital, Dallas, Texas, USA CONCLUSIONS: This study demonstrates that jSpA patients exhibit complex patterns of gene expression for functions related to inflammatory and defense re- OBJECTIVE: To evaluate the response to IL-1 blockade over the past 7 years sponse, MAP kinase and cell cycle, chromatin modulation and transcription, cell in SOJIA patients treated with anakinra at the clinical level including the durabil- death, apoptosis, and interestingly, gene closely linked to autoinflmmatory diseases ity of response, long term complications, and steroid sparing effect as well as to (NRLP3). utilize blood gene expression profiling for insight into potential mechanisms of LITERATURE: 1. Barnes MG, Aronow BJ, Luyrink LK, Moroldo MB, Pavlidis pathogenesis. P, Passo MH, Grom AA, Hirsch R, Giannini EH, Colbert RA, Glass DN, Thompson METHODS: Clinical/laboratory data of all children with SOJIA treated with ana- SD. Gene expression in juvenile arthritis and spondyloarthropathy: pro-angiogenic kinra at our institution with at least 6 months of follow-up were reviewed. Whole ELR+ chemokine genes relate to course of arthritis. Rheumatology (Oxford). 2004 blood gene expression profiling (Ilumina bead chip array) were obtained in a subset Aug;43(8):973-9. Epub 2004 May 18. of 12 patients before and after initiation of IL-1 blockade. 2. Harjacek M et al. HLA-B*27/HLA-B*07 in combination with D6S273-134 al- RESULTS: 31 SOJIA patients (18F/13M) with median disease duration of 2.6 lele is associated with increased susceptibility to juvenile spondyloarthropathies. years (range 0 days post dx-11.6yrs) and median of 3 active joints (range 0-34) Clin Exp Rheumatol. 2008 May-Jun;26(3):498-504. at initiation of anakinra were treated with a mean dose of 2.34 mg/kg (range 1.33- 5.95) with an average follow-up of 4.16 years (range 0.49-6.79)on anakinra. All children had a SOJIA signature as previously described (1)by microarray analysis. After IL-1 blockade, significant improvements were seen in rash (p=0.0008), fever (p<0.0001), number of active joints (p<0.0001), WBC (p<0.0001), hemoglobin O 02 (p,0.0001), platelets (p=0.0004), and ESR(p<0.0001). Autoinflammatory gene polymorphisms and susceptibility to UK Two patients with clinical MAS at initiation of anakinra had complete resolution. juvenile idiopathic arthritis: association with an exonic single Pre-anakinra 14 patients (46%)received IVMP (the primary method of steroid treat- nucleotide polymorphism in mevalonate kinase ment at our institution)with only 2(7%)still receiving it 6 months post anakinra (p=0.0005). A Hinks, P Martin, TG Day, J Packham, Childhood Arthritis Prospective Study 25/31 (81%) patients met Wallace criteria for clinical remission off medications. (CAPS), BSPAR study group, W Thomson, AV Ramanan & RP Donn Four children (13%) had a partial response with important clinical improvements Musculoskeletal Research Group, University of Manchester, England, UK. and were able to stop or greatly wean steroids. Two children (6%) had no sustained response, one of whom took anakinra for less than 6 weeks due to painful injec- BACKGROUND: Autoinflammatory syndromes, also called hereditary periodic tions. At last clinic visit 12 patients were on anakinra monotherapy, 1 on anakinra was fever syndromes, are a heterogeneous group of diseases, unified by a common fea- tolerating a prednisone wean (3mg/d), 3 were on anakinra/MTX,5 patients were on ture of cyclical episodes of unexplained inflammation and fever. Autoinflammatory no medications. Two patients who stopped anakinra later flared with arthritis only syndromes have several overlapping features with JIA, especially with systemic- and were controlled with etanercept. One patient developed hepatitis and had to onset JIA. We have previously studied SNPs in 4 genes that cause autoinflamma- stop anakinra. Six new onset patients and 4 polycyclic patients with new disease tory diseases (NLRP3, NOD2, MEFV, and PSTPIP1) and shown association with flares have been successfully treated with anakinra monotherapy. psoriatic onset JIA (Day TG et al. 2008, Arth & Rheum, 58(7), 2142-6). Here we Most side effects were minor although 1 patient who had just received IVMP died have studied SNPs across MVK, located on chromosome 12q24, responsible for of complications related to sepsis. One patient continued anakinra throughout preg- hyper-IgD syndrome (HIDS), and TNFRSF1A, located on chromosome 12p13.2, nancy and delivered a normal term baby. responsible for TNF receptor-associated periodic syndrome (TRAPS). In addi- Gene expression profiling showed a remarkably homogeneous pattern of IL-1 re- tion, we studied SNPs across NALP1, a homologue of the autoinflammatory gene lated gene dysregulation (1) which normalized in the most patients after anakinra. NALP3. SNPs in NALP1 have recently been associated with vitiligo-associated In some patients there was a time lapse between clinical response and correction autoimmune disease, autoimmune Addison’s disease, and type 1 diabetes. of gene expression suggesting that immune alterations are not completely resolved METHODS: DNA was available for 1054 UK Caucasian JIA patients. Pair-wise at time of first clinical response. Most upregulated transcripts encoded innate im- tagging SNPs were selected within 10kb up and down stream of each gene us- munity related proteins.Down regulated transcripts encoded proteins involve in ing an r2 cutoff ≥ 0.8 and MAF ≥ 0.05. SNP genotyping was performed using cytoxic/NK cellfunction and protein synthesis. In this cohort, interferon inducible the Sequenom iPlex® MassARRAY platform according to manufacturers instruc- genes were upregulated especially in patient who had never received steroids and tions. A 90% sample quality control rate and 90% SNP genotyping success rate was 5/7 of these showed normalization post anakinra. imposed on the analysis. Control samples genotype data was available from the CONCLUSIONS: Anakinra is safe and effective in controlling clinical disease Wellcome Trust case control consortium 2 (WTCCC2) (n=5380). Genotype and and correcting gene expression alterations in most children with SOJIA both in allele frequencies were compared between cases with JIA and controls using the combination with methotrexate and or steroids and as monotherapy.