AB0040 PYRUVATE DEHYDROGENASE KINASES AS A POTENTIAL TARGET IN THE TREATMENT OF OSTEOARTHRITIS TO UNLEASH THE METABOLIC FLOW?

Abstract

Background:While osteoarthritis (OA) is the most common joint disease worldwide, rheumatoid arthritis (RA) represents the most common type of autoimmune arthritis. In both diseases, fibroblast-like synoviocytes (FLS), which maintain the structural and dynamic integrity of the joint, have been identified as key drivers of cartilage degradation. FLS can be divided into two major populations. The destructive phenotype which is restricted to the THY1- FLS of the synovial lining promotes bone erosion, while THY1+ FLS of the sublining layer drives synovitis. The FLS phenotype is shaped by glucose metabolism, which promotes disease progression in patients with synovitis. However, profound knowledge about the contribution of FLS to pathogenic mechanisms in cartilage degradation is limited.Objectives:Here, we present the phenotypic features of FLS obtained from patients with OA (OA-FLS) compared to bone marrow-derived mesenchymal stromal cells (MSC) on transcriptomic, proteomic and metabolic levels with the aims (i) to identify novel targets for the development of disease-modifying osteoarthritis drugs and (ii) to distinguish both cell types.Methods:To this end, we comprehensively compared human bone marrow-derived MSC with OA-FLS isolated from human knee joint sections. MSC and OA-FLS were characterized in detail according to their multipotency, surface marker pattern, cell viability, proliferation rate, morphology and expression of fibroblast- and metabolic-related markers using flow cytometry, immunofluorescence and SeahorseTM. More in-depth, selected gene and protein expression patterns were analyzed using qPCR and mass spectrometry.Results:We observed a similar phenotype of OA-FLS and MSC with regard to the minimal criteria that define a MSC phenotype. In-depth comparison of OA-FLS and MSC on proteome level revealed 598 differentially expressed proteins. We observed no differences in the expression of classical fibroblast markers such as vimentin, tenascin C and decorin as confirmed on RNA level. Remarkably, fibronectin, which is mainly produced by fibroblasts, is significantly lower expressed at both protein and RNA levels in OA-FLS together with collagen type 1 and CD106. Conversely, CD9, CD54 and fibroblast-specific protein-1 were expressed significantly higher in FLS at both levels, while hyaluronan synthase 1-3 remained unchanged. Of note, in terms of mitochondrial function, human OA-FLS show a significantly lower basal respiration and ATP production than MSC, but a comparable spare respiratory capacity and cellular mitochondrial dehydrogenase activity (NADH amount) per cell. Additionally, we identified the pyruvate dehydrogenase kinase (PDK) 3 to be highly expressed in OA-FLS, while the expression of mitochondrial ATP synthase subunits, electron transport chain complexes and glycolytic enzymes was comparable with MSC. Finally, inhibition of PDK by using DCA resulted in a significant increase in oxygen consumption rate and ATP production in OA-FLS. Thus, our data newly suggest, that PDKs may play a crucial role in the pathogenesis of OA and possibly RA.Conclusion:Our data provide evidence that, although the classical fibroblast markers do not discriminate between MSC and FLS, the latter demonstrate a significantly higher expression of PDKs, known to inhibit the pyruvate entry into the TCA cycle which finally limits the mitochondrial ATP production. Therefore, shifting the metabolism of FLS from glycolysis to mitochondrial respiration via inhibition of PDKs might be a novel approach in OA for the development of disease-modifying osteoarthritis drugs in order to unleash the metabolic flow.Disclosure of Interests:None declared