A proteomic investigation of inflammatory status before and after treatment of osteoarthritis with gold microparticles, and correlation to rheumatoid arthritis

Abstract

Introduktion: Artrose er en bred klinisk betegnelse der inkluderer mange forskellige sygdomme med forskellig etologi. Den mest almindelige type er osteoartrose, sm behandles med NSAIDer og glukkokortikoider for at kontrollerer smerte og mindske inflammation. Men de fleste behandlingsmuligheder er systemiske med mulighed for svære systemiske bivirkninger. En ny mulig behandlingsform i stand til at undgå det er intraartikulære guld mikropartikel implantater, som formodes at inhiberer artikulær inflammation på flere måder, som inhibering af NF-kB signaleringsvejen. Den mest almindelige autoimmune type af artrose er rheumatoid artrit, men trods en forskellig patogenese fra OA spiler inflammation også er stor rolle i RA, og hovedformålet med behandlingen er også her at mindske den inflammatoriske status og kontrollere smerte. Som ved OA blive både NSAIDer og glukkokortikoider også brugt i behandlingen af RA, sammen med DMARDs og bDMARDs som alle har potentielle systemiske bivirkninger. Formål: Formålet med dette studie er ar klarlægge den formodede virkningsmekanisme og antiinflammatoriske potentiale af guld mikropartikler som en behandling af OA. Proteomisk profilering af synovialvæske og serumprøver før og efter behandling blev udført for at undersøge denne nye behandlings effekt på inflammation. Et andet formål er at lede efter biomarkører i guldbehandlede patienter for at monitorere behandlingens effekt, for at muliggøre en personaliseret tilgang til guld behandling. Derudover er serumprøver, før og efter behandling af RA med etanercept, også analyseret for at lede efter fælles træk i inflammatorisk status der kunne foreslå en mulig positiv effekt af guldpartikler i behandlingen af RA. Metode: Parrede biologiske prøver fra 29 OA-patienter blev indsamlet før- og 8 uger efter behandling med guldpartikler. Disse blev analyseret med kvantitativ proteomics, Innate immunregulatoriske cfDNA-målinger, og cytokinprofilering for at evaluerer inflammatorisk status før og efter behandling. Parrede biologiske prøver fra 18 RA-patienter før og efter behandling med etanercept blev også analyseret med proteomics og cfDNA-målinger og sammenlignet med resultaterne efter OA guld-behandling. Resultater: 24 immunologiske molekyler blev identificeret med PEA til at være signifikant nedreguleret som følge af en OA-guldbehandling. Flere af disse har direkte forbindelse til både OA, RA, inflammation go NF-kB signaleringsvejen. MS identificerede 22 signifikant regulerede proteiner i OA, hvoraf flere også kan kædes sammen med OA, RA, inflammation og NF-kB. cfDNA viste sig ikke at blive signifikant ændret efter otte ugers guldbehandling, men andre studier og observationer indikerer at otte uger måske ikke er nok tid til at guldbehandlingen kan udøve sin fulde effekt. Konklusion: De klinisk observerede effekter af OA guldbehandlingen fandt basis i de observerede proteomiske ændringer identificeret i dette studie, i forhold til både mindsket inflammatoriske markører, modulation af NF-kB og sammenhæng med OA-patologi. Disse effekter koblet med den proteomiske data og viden om RA patogenese, åbner for muligheden om en positiv effekt ved guld mikropartikel behandling af RA.

Introduction: Arthritis is a broad clinical term that includes many different joint diseases of different etiology. The most common type is osteoarthritis, which is treated with NSAIDs and glucocorticoids to manage pain and dampen inflammation. However, most current treatment options are systemic with potential systemic adverse effects. A new possible treatment to overcome this is intra articular gold microparticle implants which is thought to inhibit articular inflammation in several ways, sch as inhibition of the NF-kB pathway. The most common autoimmune type of arthritis is rheumatoid arthritis, and despite a different pathogenesis than OA, inflammation also plays a major role in in RA, and the main goal of treatment is also here to decrease the inflammatory status and manage pain. Like in OA, NSAIDs and glucocorticoids are used in RA treatment along with DMARDs and bDMARDs which all have potential systemic adverse effects as well. Aim: The aims of this study are to elucidate the supposed mode of action and anti-inflammatory potential of gold microparticles as a treatment of OA. Proteomic profiling of synovial fluid and serum samples before and after treatment were performed to investigate this novel treatment’s impact on inflammation. Another aim is to search for biomarkers in gold treated patients to monitor treatment effect, and to enable the possibility for a personalized treatment. Furthermore, serum samples before and after RA treatment with etanercept will also be analyzed to look for similarities in inflammatory status that could suggest a possible effect of gold particles in the treatment of RA. Methods: Paired biological samples from 29 OA patients were collected pre- and 8 weeks post treatment with gold particles. These were analyzed using quantitative proteomics, innate immune regulatory cfDNA-measurement, and cytokine profiling to evaluate inflammatory status before and after treatment. Paired biological samples from 18 RA patients before and after treatment with etanercept were also analyzed with proteomics and cfDNA-measurement as well and correlated to the results of OA gold treatment. Results: 24 immunologically associated molecules identified by PEA were significantly downregulated following gold treatment of OA. Several of these have direct correlations to both OA, RA, inflammation, and the NF-kB pathway. MS found 22 proteins to be significantly regulated in OA, several of which can be correlated with OA, RA, inflammation, and NF-kB as well. cfDNA did not show a significant drop following eight weeks of OA gold treatment, but other studies and observations indicate that 8 weeks might not be enough time to elicit the full effect of a gold treatment. Conclusions: The clinically observed effects of an OA gold treatment found basis in the observed proteomic changes identified in this study, in respect to both decreased inflammatory markers, modulation of NF-kB and connection to OA pathology. These effects coupled with the proteomic data and knowledge of an RA pathogenesis, opens the possibility of positive effects by gold microparticle treatment of RA as well.

A master thesis from Aalborg University

A proteomic investigation of inflammatory status before and after treatment of osteoarthritis with gold microparticles, and correlation to rheumatoid arthritis