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.
