Molecular aspects of musculoskeletal diseases and the role of small regulatory RNAs

Revmatologická onemocnění bývají chronická, bolestivá a do určité míry invalidizující. Porozumění mechanismům jejich patogeneze je zatím velmi neúplné. Hyper-reaktivní imunitní systém a porucha autotolerance jsou zřejmě doplněny lokálními faktory, které způsobí, že některé klouby/svaly bývají postižené více a jiné méně. Vše je výsledkem složité sítě interakcí mezi imunitními buňkami, kloubními fibroblasty, chondrocyty, osteocyty, myocyty a dalšími buňkami. V předložené dizertační práci jsem se zaměřila na tři molekulární aspekty patogeneze revmatologických onemocnění: regulační RNA, S100 proteiny a autoprotilátky. V teoretické části jsem shrnula, co je známo o vzniku, funkci a významu těchto molekul v revmatologii. Ve výzkumné části předkládám šest originálních publikací a jedno review o roli těchto molekul při vývoji revmatoidní artritidy (RA) a idiopatické zánětlivé myopatie (IZM). Mezi hlavní výsledky práce patří studie popisující expresi PIWI-interagujících RNA (piRNA) v synoviálních fibroblastech pacientů s RA. piRNA jsou malé regulační RNA, které v komplexu s PIWIL proteiny regulují genovou expresi a tlumí transpozómy. Exprese piRNA molekul byla dlouho považována za výhradní vlastnost zárodečných a nádorových buněk. My jsme detekovali 268 piRNA molekul v RA synoviálních fibroblastech (SF), a...Rheumatic diseases are common, usually chronic, painful and to some extent invalidating medical conditions. Understanding of the disease pathogenesis is still very fragmentary. Hyperreactivity of the immune system and defect of autotolerance are probably contributed by local factors, which helps to explain, why some joints/muscles are more affected than others. All this results from a complex net of interactions between immune cells, synovial fibroblasts, chondrocytes, osteocytes, myocytes and other cells. In the submitted PhD thesis I have focused on three groups of molecules: regulatory RNAs, S100 proteins and autoantibodies. In the theoretical part, I sum up the current knowledge on their biogenesis, function and the role in rheumatology. In the investigative part, I present six original publications and one review on the role of those molecules in development of rheumatoid arthritis (RA) and idiopathic inflammatory myositis (IIM). One of the main studies was focused on expression of PIWI-interacting RNAs (piRNAs) in RA synovial fibroblasts (SF). piRNAs are small regulatory RNAs which in complex with PIWIL proteins regulate gene expression and silence transpozoms. piRNA expression was considered to be limited to germline and cancer cells. We have found 267 PIWI-interacting RNAs to be expressed...Revmatologická klinika 1. LF UK a Revmatologický ústavDepartment of Rheumatology First Faculty of Medicine and Rheumatology InstituteFirst Faculty of Medicine1. lékařská fakult

Increasing incidence of immune-mediated necrotizing myopathy:single-centre experience

OBJECTIVES: Immune-mediated necrotizing myopathy (IMNM) is characterized by the predominant presence of necrotic muscle fibres in muscle biopsy and variable response to immunosuppressive treatment. The aims of this study were to analyse the temporal trend of IMNM incidence in our centre over the past 10 years and to explore the role of statins as possible causative agents.METHODS: A retrospective evaluation of muscle biopsy results, clinical and laboratory data, including antibody associations of all patients with idiopathic inflammatory myopathy newly diagnosed between 2004 and June 2014, was performed. Available sera were tested for the presence of anti-3-hydroxy-3-methylglutaryl coenzyme A reductase (anti-HMGCR) autoantibodies.RESULTS: Of 357 biopsied patients, 233 fulfilled criteria for inflammatory/immune-mediated myopathy, including 27 (11.6%) classified as IMNM. There were no patients with IMNM diagnosed between 2004 and 2007; subsequently, two to three cases of IMNM per year were seen during the period 2008-11, with a substantial increase to 18 cases (66.6% of all IMNM biopsies) in 2012-14. Thirteen of 27 patients (48%) had a history of statin use, 11 (85%) of whom had positive anti-HMGCR antibodies. There was no IMNM patient without a history of statin use who was anti-HMGCR antibody positive.CONCLUSION: Our data show an increasing incidence of IMNM, which is mainly accounted for by anti-HMGCR-positive IMNM associated with the use of statins.</p

Molecular aspects of musculoskeletal diseases and the role of small regulatory RNAs

Rheumatic diseases are common, usually chronic, painful and to some extent invalidating medical conditions. Understanding of the disease pathogenesis is still very fragmentary. Hyperreactivity of the immune system and defect of autotolerance are probably contributed by local factors, which helps to explain, why some joints/muscles are more affected than others. All this results from a complex net of interactions between immune cells, synovial fibroblasts, chondrocytes, osteocytes, myocytes and other cells. In the submitted PhD thesis I have focused on three groups of molecules: regulatory RNAs, S100 proteins and autoantibodies. In the theoretical part, I sum up the current knowledge on their biogenesis, function and the role in rheumatology. In the investigative part, I present six original publications and one review on the role of those molecules in development of rheumatoid arthritis (RA) and idiopathic inflammatory myositis (IIM). One of the main studies was focused on expression of PIWI-interacting RNAs (piRNAs) in RA synovial fibroblasts (SF). piRNAs are small regulatory RNAs which in complex with PIWIL proteins regulate gene expression and silence transpozoms. piRNA expression was considered to be limited to germline and cancer cells. We have found 267 PIWI-interacting RNAs to be expressed..

Family psychoeducation and relapse prevention in psychotic disorders

Relapse prevention is a primary long-term clinical goal in treatment of psychotic disorders. Pharmacotherapy by itself is not sufficient, because compliance of patients in remission is usually low and there are more factors contributing to relapse, especially stress of patients resulting from emotionally demanding situations in the family. PREDUKA (PREventive EDUcational programme for relapse prevention) is a six-hour professionally-led group programme for patients with psychotic disorders in ambulant therapy and for their relatives. In a field questionnaire survey we mapped 1) theoretical knowledge of participants, 2) usage of practical advice obtained from PREDUKA and 3) general benefit of programme. Participants from years 2007 - 2009 were included. The sample consisted of 14 patients and 22 relatives, 27 women and 11 men. The average age of patients was 28,6 years, that of relatives 44,4 years. High return of questionnaires from patients confirms their goodwill to cooperate. Knowledge of participants about psychosis and their therapy is very good, compliance of patients from our sample is high. Participants usually follow advice obtained from PREDUKA. Common is the ignorance of important phone numbers (crisis centre, attending psychiatrist), so we suggest to include a page in a workbook for these dates...

Molecular aspects of musculoskeletal diseases and the role of small regulatory RNAs

Rheumatic diseases are common, usually chronic, painful and to some extent invalidating medical conditions. Understanding of the disease pathogenesis is still very fragmentary. Hyperreactivity of the immune system and defect of autotolerance are probably contributed by local factors, which helps to explain, why some joints/muscles are more affected than others. All this results from a complex net of interactions between immune cells, synovial fibroblasts, chondrocytes, osteocytes, myocytes and other cells. In the submitted PhD thesis I have focused on three groups of molecules: regulatory RNAs, S100 proteins and autoantibodies. In the theoretical part, I sum up the current knowledge on their biogenesis, function and the role in rheumatology. In the investigative part, I present six original publications and one review on the role of those molecules in development of rheumatoid arthritis (RA) and idiopathic inflammatory myositis (IIM). One of the main studies was focused on expression of PIWI-interacting RNAs (piRNAs) in RA synovial fibroblasts (SF). piRNAs are small regulatory RNAs which in complex with PIWIL proteins regulate gene expression and silence transpozoms. piRNA expression was considered to be limited to germline and cancer cells. We have found 267 PIWI-interacting RNAs to be expressed..

Decreased circulating visfatin is associated with improved disease activity in early rheumatoid arthritis: data from the PERAC cohort.

ObjectiveTo evaluate circulating visfatin and its relationship with disease activity and serum lipids in patients with early, treatment-naïve rheumatoid arthritis (RA).MethodsSerum visfatin was measured in 40 patients with early RA before and after three months of treatment and in 30 age- and sex-matched healthy individuals. Disease activity was assessed using the Disease Activity Score for 28 joints (DAS28) at baseline and at three and 12 months. Multivariate linear regression analysis was performed to evaluate whether improved disease activity is related to serum visfatin or a change in visfatin level.ResultsSerum visfatin was significantly elevated in early RA patients compared to healthy controls (1.92±1.17 vs. 1.36±0.93 ng/ml; p = 0.034) and significantly decreased after three months of treatment (to 0.99±0.67 ng/ml; pConclusionA short-term decrease in circulating visfatin may represent an independent predictor of long-term disease activity improvement in patients with early RA

Expression and Regulation of PIWIL-Proteins and PIWI-Interacting RNAs in Rheumatoid Arthritis.

The PIWIL (P-element induced wimpy testis like protein) subfamily of argonaute proteins is essential for Piwi-interacting RNA (piRNA) biogenesis and their function to silence transposons during germ-line development. Here we explored their presence and regulation in rheumatoid arthritis (RA).The expression of PIWIL genes in RA and osteoarthritis (OA) synovial tissues and synovial fibroblasts (SF) was analysed by Real-time PCR, immunofluorescence and Western blot. The expression of piRNAs was quantified by next generation small RNA sequencing (NGS). The regulation of PIWI/piRNAs, proliferation and methylation of LINE-1 after silencing of PIWIL genes were studied.PIWIL2 and 4 mRNA were similarly expressed in synovial tissues and SF from RA and OA patients. However, on the protein level only PIWIL4 was strongly expressed in SF. Using NGS up to 300 piRNAs were identified in all SF without significant differences in expression levels between RA and OASF. Of interest, the analysis of the co-expression of the detected piRNAs revealed a less tightly regulated pattern of piRNA-823, -4153 and -16659 expression in RASF. In RASF and OASF, stimulation with TNFα+IL1β/TLR-ligands further significantly increased the expression levels of PIWIL2 and 4 mRNA and piRNA-16659 was significantly (4-fold) induced upon Poly(I:C) stimulation. Silencing of PIWIL2/4 neither affect LINE-1 methylation/expression nor proliferation of RASF.We detected a new class of small regulatory RNAs (piRNAs) and their specific binding partners (PIWIL2/4) in synovial fibroblasts. The differential regulation of co-expression of piRNAs in RASF and the induction of piRNA/Piwi-proteins by innate immune stimulators suggest a role in inflammatory processes