1. Översiktlig projektbeskrivning

Engelsk titel

The role of oncogenes as initiators and therapeutic targets in joint inflammation

Sammanfattning av projektet

Inflammation har en central roll vid uppkomsten och progression av tumörer liksom vid autoimmuna sjukdomar. De intracellulära processer som startar vid inflammation leder till påverkan på cellcykeln, apoptos defekter och kan kulminera i malign cellomvandling.
Den föreliggande ansökan syftar till att skapa förståelse hur den kroniska, invasiva, inflammatoriska processen vid reumatoid artrit sker. Denna process har flera likheter med tumörspridning, genom synovialvävnadens aggressiva växtsätt, okontrollerad spridning till kroppen alla leder liksom frånvaro av effektiv apoptos. Vi har tidigare visat att anti-apoptotisk protein survivin som har central betydelse för utveckling och spridning av cancermetastaser är uppreglerad i RA. Survivin har prediktivt värde för att identifiera patienter i risk att utveckla leddestruktioner.
Målet är att värdera relationen mellan inflammation vid artrit och förändringar i ledvävnadsceller avseende modulation av survivinsyntes, S100A4 reglering av benomsättning och T cellsutveckling, och ingripande i dessa mekansimer genom KO möss och siRNA teknologi.
Dessa mål kan nås genom att studera intracellulära mekanismer resulterade i överuttryck av survivin och S100A4 och dess påverkan på immunsystemet. Vi kommer studera inverkan av ovan proteiner för differentiering till osteoklaster, benförstörande celler, vid RA. Vi planerar att värdera in vivo oortodox proteinexpression för att modulera det specifika respektive generella inflammatoriska svaret.
Våra preliminära resultat tyder på att proto-onkogener survivin och S100A4 är av stor betydelse för utveckling av en förstörande ledsjukdom och kan övervägas som mål för en riktad terapi vid RA.

Typ av projekt

Forskningsprojekt

MeSH-termer för att beskriva ämnesområdet

Inlagda MeSH-termer

Musculoskeletal Diseases

Diseases of the muscles and their associated ligaments and other connective tissue and of the bones and cartilage viewed collectively.

Projektets delaktighet i utbildning

Avhandling
D-uppsats / Magisterexamen
C-uppsats / Kandidatexamen
ST-läkarutbildning
Annan utbildning
Ej del i utbildning

3. Processen och projektets redovisning

Pågående aktiviteter

Planering och förberedelse före datainsamling
Datainsamling pågår
Analys av insamlade data pågår
Författande av skriftlig redovisning / publikation pågår
En eller flera publikationer från projektet är publicerade
Slutfört och inget mer görs inom ramen för detta projekt

Projektstart (när planeringen påbörjas och börjar dokumenteras skriftligt)

2008-01-01

Datum för påbörjande av datainsamling

2008-06-01

Datum då projektet är slutrapporterat

2011-12-31

Publikationer från detta projekt

1. Resistin competes with lipopolysaccharide for binding to Toll-like receptor 4.
   Tarkowski Andrej, Bjersing Jan, Shestakov Andrey, Bokarewa Maria I.
   J Cell Mol Med. 2009[Source: PubMed®]
   [Links: PMID: 19754671 | DOI länk]
2. Arthritogenic dsRNA is present in synovial fluid from rheumatoid arthritis patients with an erosive disease course.
   Bokarewa Maria, Tarkowski Andrej, Lind Magnus, Dahlberg Leif, Magnusson Mattias.
   Eur J Immunol. 2008:38(11):3237-44.[Source: PubMed®]
   [Links: PMID: 18991285 | DOI länk]
3. Single-stranded polyinosinic acid oligonucleotides trigger leukocyte production of proteins belonging to fibrinolytic and coagulation cascades.
   Zare Fariba, Magnusson Mattias, Möllers Linda Nilsson, Jin Tao, Tarkowski Andrej, Bokarewa Maria.
   J Leukoc Biol. 2008:84(3):741-7.[Source: PubMed®]
   [Links: PMID: 18562486 | DOI länk]
4. Role of resistin as a marker of inflammation in systemic lupus erythematosus.
   Almehed Katarina, d'Elia Helena Forsblad, Bokarewa Maria, Carlsten Hans.
   Arthritis Res Ther. 2008:10(1):R15.[Source: PubMed®]
   [Links: PMID: 18234104 | DOI länk]
5. Extracellular survivin up-regulates adhesion molecules on the surface of leukocytes changing their reactivity pattern.
   Mera Simona, Magnusson Mattias, Tarkowski Andrej, Bokarewa Maria.
   J Leukoc Biol. 2008:83(1):149-55.[Source: PubMed®]
   [Links: PMID: 17938276 | DOI länk]
6. Arthritogenic properties of double-stranded (viral) RNA.
   Zare Fariba, Bokarewa Maria, Nenonen Nancy, Bergström Thomas, Alexopoulou Lena, Flavell Richard A, Tarkowski Andrej.
   J Immunol. 2004:172(9):5656-63.[Source: PubMed®]
   [Links: PMID: 15100310]
7. Phosphorotioated oligonucleotides trigger synthesis of human coagulation serine proteases.
   Bokarewa Maria I, Hellstrand Kristoffer, Tarkowski Andrej.
   Thromb Haemost. 2002:88(5):788-93.[Source: PubMed®]
   [Links: PMID: 12428095 | DOI länk]
8. Resistin exerts its proinflammatory effects through interaction with Toll-like receptor 4.
   Bokarewa Maria, Bjersing Jan, Nagaev Ivar, Shestakov Andrej, Isaksson Olle, Smith Ulf, Tarkowski Andrej.
   2008[Source: User]

4. Detaljerad projektbeskrivning

Bakgrundsbeskrivning

In most cases when a few normal cells evade the normal cell cycle control and begin to divide uncontrollably, an inflammatory response can mount an attack and eliminate the early cancer cells. However, the inflammatory cells can also contribute to tumor development. Epidemiological studies have established a strong correlation between inflammation and a predisposition for cancer. Already in the 19th century, Virchow noted that many tumors show a “lymphoreticular infiltrate”. Today, it is well known that inflammatory cells are present in tumors and can contribute to the development and progression of cancer.

Rheumatoid arthritis (RA) is a chronic and destructive autoimmune disease, which affects almost 1% of the world population and occasionally ends-up in malignant lymphoproliferative diseases such as lymphomas. Uncontrolled, tumor-like growth of synovial tissue, followed by invasion and destruction of cartilage is a classical feature of RA. Synovial fibroblasts of RA patients have the ability to proliferate and to invade the surrounding tissues in a tumor-like fashion. This excessive growth of rheumatoid synovium, resembling tumour tissues, displays an elevated expression of proto-oncogenes such as c-Myc, c-Ras, c-Jun and low activity of tumor suppressor genes p53 and PTEN (2). At present, very little is known about whether expression of an oncogene in the inflammatory cells themselves (e.g., fibroblasts, myeloid cells, lymphocytes) contributes to the development or progression of RA.

Survivin is a multifunctional protein that serves as an efficient inhibitor of apoptosis, and a regulator of cell division. Most knowledge about survivin has accumulated with respect to tumorigenesis. Survivin is abundantly expressed in most tumors being frequently used to predict efficiency of treatment and overall survival in malignancies. We have recently shown that high extra-cellular levels of surivin may be used as an independent marker predicting development of destructive RA.

Tyrosine kinases are cell surface or intracellular enzymes regulating cell proliferation, cytoskeletal reorganization and apoptosis. Activation of tyrosine kinases induces intracellular cascade of reactions involving Ras/Raf/MAP-kinase pathway or phosphatidylinositol 3-kinase /Akt pathway. Interaction between intracellular pathways activated by inflammation and by tyrosine kinases is not completely clear. It has been recently shown that expression of an oncogenic form of K-Ras (Gly12Asp) in bronchiolar epithelium in mice does not induce lung cancer but instead results in a massive inflammatory response with infiltration of alveolar macrophages and neutrophils and an increased expression of pro-inflammatory cytokines. This study, and others, establishes that inflammation can be elicited in the setting of oncogene expression in the lung. Other studies show that another oncogene, S100A4, is highly expressed in inflammatory cells in the joint of patients with RA.

Syfte

To evaluate possible link between inflammation processes in rheumatoid arthritis and tumor-like changes in synovial tissues regulated by survivin and S100A4 protein.

Frågeställning / Hypoteser

Survivin and S100A4 play important role in joint destruction induced by rheumatoid arthritis

Metod: Databearbetning

Statistical evaluation of the results will be performed using mann-Whitney statistics for comparison between the groups. Within the group the results will be compared by Student paired t-test

Resultat

1. We have recently shown that high extracellular levels of proto-oncogene survivin are detected in circulation of 30-50% of RA patients (1). Moreover, data obtained in the prospective follow-up of Swedish patients with early stage of RA (BARFOT cohort) indicate survivin as a strong predictive marker for the development of destructive RA (9). We have also shown that suppression of survivin gene using siRNA construct in fibroblasts results in prevention of arthritis induced by instillation of unmanipulated fibroblasts in the knee joint of healthy mice (8).
2. We have also evaluated mechanisms stimulating excessive expression of survivin in RA patients. We observed that intracellular pathways regulated by tyrosine kinase and PI3-kinase controls survivin expression in human leukocytes and in synovial fibroblast cell cultures (8). We have recently applied synthetic inhibitors of tyrosine kinases (SU11248) in the mouse models of methylated-BSA induced arthritis. A significant reduction in arthritis index evaluated histologically was observed. Additionally, inhibition of tyrosine kinases in vivo had immunomodulating properties for antigen presentation resulted in a reduction in mBSA-specific and aCCP antibody production (manuscript in preparation 14).
3. In collaboration with a group in Denmark we have begun to define the role of the oncogene S100A4 in the development of joint inflammation. Preliminary data suggest that in serum and synovial fluid from patients with RA there are high levels of a soluble form of the S100A4 oncogene bound as an immune complex to IgM. Such immune complexes are known to trigger profound inflammation in vivo. In this project, we will determine to what extent and by what mechanism the S100A4-IgM immune complexes can trigger joint inflammation.
4. We also found that S100A4-deficiency is associated with excessive bone formation (osteopetrosis). These findings in the first preliminary experiment are now further studied following bone formation in castrated male and female mice. Markers of bone metabolism are now studied in vitro. We now develop in vitro system for differentiation of bone progenitors (osteoclasts, bone resorbing cells and osteoblasts, bone forming cells) in survivin-rich millieu using human and mouse bone marrow cells of unmanipulated and S100A4-deficient mice.

Referenser

Original publications relevant for the project
M.Bokarewa - The role of oncogenes as initiators and theraputic targets in joint inflammation

1.Bokarewa MI, Lindblad S, Bokarew D, Tarkowski A. 2005. Increased expression of survivin, an anti-apoptotic protein, in synovial fluid of patients with rheumatoid arthritis. Arth Res Ther 7, R349-R358 (http://arthritis-research.com/content/7/2/R349).
2.Jin T, Bokarewa MI, Tarkowski A. 2005. Urokinase-type plasminogen activator, an endogenous antibiotic. J Infect Dis 192, 3:429-37.
3.Jin T, Bokarewa MI, Tarkowski A. 2005. The role of urokinase in innate immunity against Staphylococcus aureus. Microb Infect 7, 9-10:1170-5.
4.Bokarewa MI, Dahlberg L, Tarkowski A. 2005. Production of antibodies against tissue inhibitors of metalloproteinases (TIMP) may prevent bone destruction in patients with rheumatoid arthritis. Arth Res&Ther 7, 5:1014-22.
5.Galeotti L, Adrian K, Berg S, Tarkowski A, Bokarewa MI. 2008. Extracellular survivin is a marker of severe juvenile chronic arthritis. Clin Exp Rheumatol 26:373-8.
6.Mera S, Magnusson M, Tarkowski A, Bokarewa MI. 2008. Extracellular survivin upregulates adhesion molecules on the surface of leukocytes changing their reactivity pattern. J Leuk Biol 83:149-55.
7.Dehlin M, Bokarewa MI, Rottapel R, Dahlberg L, Magnusson M, Foster SJ, Tarkowski A. 2008. Intra-articular Fms-like tyrosine kinase 3 ligand expression is a driving force in induction and progression of arthritis. PLoS ONE, 3(11):e3633.
8.Baran M, Nilsson Möllers L, Andersson S, Jonsson IM, Bjersing J, Ekvall AK, Tarkowski A, Bokarewa MI. 2009. Survivin is an essential regulator of joint inflammation interacting with urokinase signaling. J Cell Mol Med, published ahead of print Feb, PMID: 19298527.
9. Svensson B, Hafström I, Forslind K, Albertsson K, Tarkowski A, Bokarewa MI. 2009. Increased expression of proto-oncogene survivin predicts joint destruction in early rheumatoid arthritis. Ann Med, in press.
10. Bian L, Jonsson IM, Josefsson E, Ohlsson C, Bokarewa M, Tarkowski A, Magnusson M. 2009. Dichloroacetate alleviates development of collagen II induced arthritis in female DBA/1 mice. Arth Res&Ther, in press.
11.Verdrengh M, Bokarewa M, Ohlsson C, Stolina M, Tarkowski A. 2009. RANKL-targeted therapy inhibits bone resorption in experimental Staphylococcus aureus-induced arthritis. BONE, published ahead of print Oct 28, 2009.