224 32nd World Congress of Internal Medicine (October 24-28, 2014) WCIM 2014
PS 0686 Rheumatology
Banking of Induced Pluripotent Stem Cells Derived from Patients with Rheumatic Diseases for Research and Clinical Purposes
Seung Min JUNG1, Seung-Ki KWOK1, Sung-Hwan PARK1, Ji Hyeon JU1 The Catholic University of Korea, Seoul St. Mary`s Hospital, Korea1
Background: Patient-derived induced pluripotent stem cells (iPSC) can be a feasible resource in the fi eld of disease modelling and regenerative medicine. Proper storage and qualifi ed supply of stem cell lines are essential for facilitating pathophysiological research and clinical application. We aimed to generate a banking system of dis- ease-specifi c iPSC for future uses.
Methods: Patients who satisfi ed the relevant classifi cation criteria for diagnosis of rheumatic diseases were recruited from outpatient clinic at department of rheuma- tology, Seoul St. Mary’s hospital. Under informed consents, blood sampling and/or skin biopsy were performed for isolation of peripheral blood mononuclear cells (PBMCs) and dermal fi broblast, respectively. The primary cells were transduced with sendai virus containing Oct4, Sox2, Klf4, and c-Myc. Pluripotency of each iPSC was determined by immunostaning, real-time polymerase chain reaction (RT-PCR), and test for teratoma formation. Short-tandem repeat (STR) testing was performed to ascertain the identity the origin of iPSC lines.
Results: More than 50 clones of disease-specifi c iPSCs were reprogrammed from PB- MCs or fi broblasts of patients with 11 representative rheumatic diseases; rheumatoid arthritis, osteoarthritis, ankylosing spondylitis, systemic lupus erythematosus, primary Sjogren’s syndrome, systemic sclerosis, etc. All of the reprogrammed iPSCs expressed pluripotent markers determined by immunostaining and RT-PCR, and formed terato- mas in SCID mice. STR test confi rmed uniqueness of cell lines without cross-contam- ination. We also verifi ed the ability to recover viable iPSC colonies after thawing vials from frozen stocks.
Conclusions: We are under developing standardization of the banking process with iPSC derived from patients suffering from rheumatic diseases. The successful banking of disease-specifi c iPSC would give rise to advances in stem cell research and therapy in rheumatology fi eld.
PS 0687 Rheumatology
Generation of Cardiomyocyte from Synoviocytes of Patients with Rheumatoid Arthritis and Osteoarthritis via Induced Pluripotent Stem Cell
Seung Min JUNG1, Seung-Ki KWOK1, Sung-Hwan PARK1, Ji Hyeon JU1 The Catholic University of Korea, Seoul St. Mary`s Hospital, Korea1
Background: Cardiovascular complication is a main cause of morbidity in arthritic patients. Cardiovascular complication in rheumatoid arthritis (RA) occasionally results from infl ammatory insults and in osteoarthritis (OA) from degenerative aging process.
To differentiate into cardiomyocytes from induced pluripotent stem cells (iPSCs) of the patients with RA and OA gives an opportunity for disease modeling or cell replacement therapy in arthritic patients.
Methods: We generated iPSCs from synoviocytes of patietns with RA and OA using 4-in-1 lentiviral vector. The pleuripotency of iPSCs were checked with RT-PCR on Na- nog, Oct4, Sox2, Klf4 and with immunofl uorescence on Nanog, SSEA4, Oct4, and Tra- 1-60. Established iPSC lines were differentiated into cardiomyocyte lineages (iPSC-CMs) using standard 3D embryoid body differentiation protocols. To investigate whether cardiac cells were properly generated, immunofl uorescence and calcium imaging were performed.
Results: Total four clones of iPSC were successfully generated from synoviocytes of two RA patients and two OA patients. mRNA expression of Nanog, Oct4, ZPF-42, and Sox2 for pluripotency markers were elevated after reprogramming process. After 12 days of differentiation protocol for cardiomyocytes, RA-iPSC-CMs and OA-iPSC-CMs were generated. Immunofl uorescence study and RT-PCR were positive results proving of mature cardiomyocyte. Calcium handling image was well observed at RA/OA-iPSC- CMs.
Conclusions: To generate patient-specifi c cardiomyoctes in arthritic patient could be potential cell source of disease modeling approach or cell therapy when those patients should take “diagnosis in a dish” or cytotherapy against failing cardiomyocytes.
PS 0688 Rheumatology
Prevalence of Serological Markers in Rheumatoid Ar- thritis
Artur ZOTO1, Zamira YLLI2, Arjan HARXHI3, Brikena SELIMI4
Rheumatology, University Hospital Center, Albania1, Immunology, University Hospital Center, Albania2, In- fectious Disease, University Hospital Center, Albania3, Ophthalmology, University Hospital Center, Albania4 Background: Rheumatoid arthritis is a highly infl ammatory polyarthritis often leading to joint destruction, deformity and loss of function. Rheumatoid factor and anti-cyclic citrullinated peptide antibody are considered to be specifi c for rheumatoid arthritis.
The aim of this study was to determine the prevalence of rheumatoid factor and an- ti-cyclic citrullinated peptide antibody in Albanian patients with rheumatoid arthritis Methods: We analyzed the prevalence of rheumatoid factor and anti-cyclic citrul- linated peptide antibody in 108 patients with rheumatoid arthritis and 86 controls.
Patients were examined by immunological tests such as rheumatoid factor and an- ti-cyclic citrullinated peptide antibody.
Results: Patients with positive rheumatoid factor were 51 (47.2 %), anti-cyclic citrulli- nated peptide antibody positive were 28 ( 25.9) patients and 16 (14.8 %) patients had positive for both serological markers. None of the controls demonstrated rheumatoid factor and anti-cyclic citrullinated peptide antibody positive.
Conclusions: Rheumatoid factor is prevalent in Albanian rheumatoid arthritis patients and more sensitive than anti-cyclic citrullinated peptide antibody. For evaluation patients with suspected rheumatoid arthritis is recommended to perform rheumatoid factor and anti-cyclic citrullinated peptide antibody.
PS 0689 Rheumatology
Polymorphism Rs3804513 of Interleukin-17a Increases the Risk of Rheumatoid Arthritis and is Correlated with the Condition’s Severity in Mexican Patients
MAXIMILIANO GARCÍA1, RENE MÉNDEZ3, AIDA GALICIA6, EFRAÍN GARRIDO4, GLUSTEIN POZO3, EDGAR MORAN5, NORMA HERRERA2
Coordinación De Planeación De Infraestructura Médica-IMSS, Mexico1, División De Estudios De Pos- grado, Instituto Politécnico Nacional, Mexico2, Facultad De Medicina, FES-Iztacala, UNAM, Mexico3, Centro De Investigaciones y Estudios Avanzados (CINVESTAV), Mexico4, Hospital General De Zona No.2-IMSS, Mexico5, Hospital General De Zona 2A, Mexico6
Background: Rheumatoid Arthritis (RA) is the most common autoimmune disease worldwide. T cells and their cytokines play a key role in the etiopathogenesis of this illness. In the last two decades IL-17A has shown to play a major role in the dis- ease pathogenesis. Previous research on the SNP (Single Nucleotide Polymorphism) rs3804513 has shown a link to radiological progression in patients with RA.
Objetive: To determine the correlation between rs3804513 IL-17A SNP and the risk of developing RA, and assess the relation between rs3804513 SNP and the severity of the disease.
Methods: Oral epithelial cells were obtained as source for genetic material. The DNA was amplifi ed using Polymerase Chain Reaction. Subsequently, a Restriction Fragment Length Polymorphism was performed. Finally, in order to confi rm the SNP presence, direct sequencing of the DNA was performed. We used a case-control design.
Results: 76 patients with RA and 94 healthy subjects were included. Of the 76 pa- tients evaluated, 15% (n=12) tested positive for SNP rs3804513 vs 3 % (n=3) sub- jects in the control group. An odds ratio (OR) of 5.6 (p=0.01) for developing RA was obtained in RA patients carrying the studied SNP. Interestingly, a large percentage of patients who tested positive required 3 DMARDs to control the disease in comparison to patients who tested negative: 73% vs 32% (OR 5.6, p<0.01).
Conclusions: The SNP rs3804513 is a risk factor for developing RA. Furthermore, car- riers need more aggressive treatment to control the disease.
WCIM 2014 SEOUL KOREA 225
Poster Session
The Korean Journal of Internal Medicine Vol. 29, No. 5 (Suppl. 1)
PS 0690 Rheumatology
Freedom Struggle Against Rheumatoid Arthritis
Kedar BHIDE1 Indira Hospital, India1
Background: Evidence from 4500 BC native Americans at Tennessee, 1500 BC Papy- rus from Egypt to the fi rst description of arthritis 400 BC from Hippocrates, Galen and many others, on the way from Paracelsus, Ayurveda, Sydenham, Beauvais to Benjamin Brodies 1818s description, Dr. Garrod in 1859 named the disease` Rheumatoid Arthritis’
and the struggle began offi cially. 1896s illustrated descriptions of Bannatyne, forma- tion of ‘International Committee of Rheumatism’, Camroe coining the term ‘Rheuma- tologist’ in 1940 & Hollander in 1949 ‘Rheumatology’, the struggle got its momentum with discovery of` Rheumatoid factor’ by Dr. Waller in 1940. On the treatment front use of Willio plant extract by Hippocrates & Galen to Grehardt & Hoffman’s (1832,1897) discovery of acetyl salicylic acid opened the gates of NSAIDS, John Vane in 1971 hon- ored with Nobel Prize for his elucidation of action of NSAIDS, with addition of COX2 selective inhibitors by Simmons in 1991. Kendal, Hench and Reichstein Nobely honored for pioneering discovery and application of corticosteroids. Accidental discovery of Gold and Antimalarials as early DMARDs [disease modifying anti rheumatoid drugs]
got the struggle augmented with DR. Y. Subbarao’s development of Methotrexate and now a new breed of biological agents promise a greater freedom from pain and suf- fering of Rheumatoid Arthritis.
Methods: The literature was reviewed from 100s of sites on internet with intricate details and various text books and Journals, systematically and methodically arranging events in chronological order of discover.
Results: There is astonishing wealth of information and inspiration is achieved, which we never realize while studying in Medical College or in Practice and we really under- stand the worth of it.
Conclusions: As per William Osler`` …the knowledge which is your privilege today to acquire so early has cost others. We are all of us, debtors to our profession.’’
PS 0691 Rheumatology
A Randomized, Double-Blind, Phase 3 Equivalence Trial Comparing the Etanercept Biosimilar, Hd203, to Ref- erence Etanercept, in Combination with Methotrexate (MTX) in Korean Patients with Rheumatoid Arthritis (RA)
Sang-Cheol BAE1, Jinseok KIM2, Jung-Yoon CHOE3, Won PARK4, So-Ra LEE5, Yong-Ho AHN6 Hanyang University Seoul Hospital, Korea1, Jeju National University Hospital, Korea2, Catholic University of Daegu, Korea3, Inha University Hospital, Korea4, Clinical Development/Regulatory Affairs, Hanwha Chemical Biologics, Korea5, R&D Center, Hanwha Chemical Biologics, Korea6
Background: Etanercept is a recombinant fusion protein that blocks TNF. HD203 is a biosimilar of etanercept with demonstrated comparability across pharmacokinetics, safety and tolerability. The objectives of this study were to evaluate equivalence in effi cacy and compare safety of HD203 with reference etanercept, in combination with MTX in patients with RA. (ClinicalTrials.gov NCT01270997).
Methods: Korean patients (male or female aged =20 years) with active RA were ran- domized (1:1) to 25 mg HD203 or reference etanercept, administered subcutaneously twice weekly with MTX for 48 weeks. The primary endpoint was the proportion of pa- tients achieving ACR20 at week 24. Secondary endpoints included ACRn, DAS28, and EULAR response at week 24 and 48, safety and immunogenicity.
Results: In total, 294 patients were randomized: HD203, n=147; reference etanercept, n=147. The proportion of patients achieving ACR20 at week 24 was not signifi cantly different between HD203 and reference etanercept. Equivalent effi cacy was demon- strated within predefined margins. There were no significant differences between proportions achieving ACR20 at week 12 and 48. ACR50 and ACR70 displayed similar trends. There were no signifi cant differences between groups for ACRn, DAS28, and EULAR response. Safety set analysis (HD203, n=147; reference etanercept, n=146) re- vealed no signifi cant difference for treatment-emergent (all-causality) adverse events (AEs): HD203 76.87% vs. reference etanercept 78.08% (p=0.8040). No significant differences between HD203 and reference etanercept were observed for adverse drug reactions, serious AEs, or discontinuations due to AEs. Few patients tested positive for anti-drug antibodies.
Conclusions: The study met the primary endpoint of demonstrating equivalent effi cacy of HD203 compared to reference etanercept. HD203 was well tolerated, with a safety profi le comparable to reference etanercept in this population of patients with RA.
PS 0692 Rheumatology
Anti TNF-Alpha Therapy in Rheumatoid Arthritis Pa- tients Disease Activation with, Correlation Between Serum Level of ESR and CRP Levels
Gulcin GUNGOR OLCUM1, Guven KOC1, Fidan Canan CELIK YAGAN3, Ece YIGIT TASKIN1, Senem ERTILAV1, Hanife Serife AKTAS1, Sati Sena YILDIZ1, Demet ATAMAN TASAN4, Fatih AKDOGAN1, Sema BASAT1, Mehmet Ali USTAOGLU2
Umraniye Training and Research Hospital, Turkey1, Dr.Lutfi Kirdar Kartal Trainig and Research Hospital, Turkey2, Zeynep Kamil Womens and Children Training and Research Hospital, Turkey3, Medical Park Hospital, Turkey4
Background: RA is a chronic systemic disease manifest with multiple joint infl am- mation. Vasculitis, heart, lung disease, extra-articular symptoms can also be present.
TNF-a,IL-6 and mediators such as cytokines, have been shown to have an important role in this infl ammatory process.Anti-TNF drugs considered to be effective in pre- venting;RA,disability and joint destruction. CRP is one of the best indicators of infl am- mation though, ESR is an indirect indicator of infl ammation and ESR levels affected by age, sex, status and anemia. ESR and CRP levels in patients with RA, disease activity and has been shown to correlate with radiographic fi ndings In this study, we aim to to show the correlation between ESR, CRP levels with disease activity in patients receiv- ing anti-TNF alpha therapy.
Methods: In this Retrospective study between January 2006 to March 2010 patients diagnosed with RA were evaluated in two groups. In the study group patients re- ceiving at least one year of TNF-alpha were included where as in the control group patients only receiving DMARD were include. Only female patients were involved in both groups. There were 46 women in the study and 47 women in the control group.
Disease severity and DAS 28 score was used to determine the disease activity. For statistical analysis Number Cruncher Statistical System 2007 & PASS 2008 Statistical Software (Utah,USA) was used.
Results: In patients receiving anti-TNF alpha therapy DAS 28 scores showed statisti- cally signifi cant correlation with the ESR. A statistically signifi cance between DAS 28 and CRP were not found. In control group with only DMARD treatment, the DAS 28
