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Abstract Number: 0705

lncRNA H19 and Micro RNA 675-3p Are Altered by Visfatin During Osteogenesis

Dennis Küppers1, Lali Tsiklauri2, Marie-Lisa Hülser1, Klaus Frommer2, Stefan Rehart3, Caroline Ospelt4, Ulf Müller-Ladner5 and Elena Neumann6, 1Justus-Liebig-University Giessen, Campus Kerckhoff, Dept. of Rheumatology and Clinical Immunology, Giessen, Germany, 2Justus-Liebig-University Giessen, Campus Kerckhoff, Dept. of Rheumatology and Clinical Immunology, Bad Nauheim, Germany, 3Department of Orthopaedics and Trauma Surgery, Agaplesion Markus Hospital, Frankfurt, Frankfurt, Germany, 4University Hospital Zürich; University Hospital of Zurich; Center of Experimental Rheumatology, Department of Rheumatology, Zurich, Switzerland, 5Department of Rheumatology, Immunology, Osteology and Physical Medicine, Justus Liebig University Gießen, Campus Kerckhoff, Bad Nauheim, Germany, Bad Nauheim, Germany, 6Department of Rheumatology and Clinical Immunology, Campus Kerckhoff, Justus Liebig University Giessen, Bad Nauheim, Germany

Meeting: ACR Convergence 2020

Keywords: Inflammation, Mesenchymal stem cells, Micro-RNA, Osteoarthritis, signal transduction

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Session Information

Date: Saturday, November 7, 2020

Title: Osteoarthritis & Joint Biology – Basic Science Poster

Session Type: Poster Session B

Session Time: 9:00AM-11:00AM

Background/Purpose: Long non-coding (lnc-)RNAs are transcribed from DNA similar to mRNA. lncRNA are able to interact directly with DNA, RNA and proteins. Some lncRNAs contain micro (mi-)RNAs in their sequence. The miRNAs can be released by splicing leading to active miRNA molecules. lncRNA H19 includes two miRNAs 675-3p and -5p in its sequence. Adipose tissue derived adipokines are involved in inflammation processes as well as osteoarthritis (OA) development. The proinflammatory adipokine visfatin is able to alter osteogenic differentiation (OD) of pluripotent mesenchymal stem cells (MSCs). Visfatin reduces elastic fiber expression, increases matrix mineralization and proinflammatory factor production.

Methods: MSCs isolated from OA hip or knee bone (phMSCs) and commercially obtained healthy human hMSCs were differentiated towards osteoblasts stimulated with or without visfatin, resistin, leptin as well as TNF and Wnt/TGFβ1 pathway inhibitors. Supernatants were collected at days 2, 7, 9, 14 and 21 of OD, cell lysates at day 2, 7, 9, 14. Matrix mineralization assays were performed at day 21. lncRNA H19 and miRNA expression was evaluated by real-time PCR after miRNA and total RNA isolation. IL-6 was measured by ELISA.

Results: Visfatin increased matrix mineralization and IL-6 release (hMSC: p=0.03, phMSC: p=0.01) (1). lncRNA H19 was continuously upregulated in unstimulated controls during OD as well as with leptin or resistin. Stimulation with visfatin significantly decreased lncRNA H19 (d2 to d14 of OD, phMSC: p=0.01, h-MSC: p=0.04). TNF stimulation during OD did not lead to downregulation of H19 nor increased matrix mineralization. lncRNA H19 endogenous miRNA 675-5p was reduced in parallel with H19: increased during OD and downregulated by visfatin significantly (e.g. d14 p=0.02). However, H19 endogenous miRNA 675-3p was inversely regulated: downregulated during OD while visfatin attenuated this effect (e.g. d14 p=0.03). Altered Wnt-signaling or TGFβ1 pathway were not observed.

Conclusion: lncRNA H19 is upregulated during OD playing a regulatory role during osteogenesis. During OD, visfatin showed proinflammatory effects and increased matrix mineralization while reducing elastic fiber production. These effects were associated with a reduction of lncRNA H19, an effect not triggered by other adipokines or TNF. We demonstrated that the lncRNA H19 endogenous miRNA 675-5p was regulated in parallel to H19, whereas miRNA 675-3p was inversely regulated and increased continuously upon visfatin stimulation. These results indicate that miRNA 675-3p is released out of the lncRNA H19 sequence leading to H19 reduction representing an effector mechanism of visfatin. We hypothesize that this is a restrictive process in which either microRNA 675-5p or -3p is liberated from H19 and, in this setting, of miRNA 675-3p.


Disclosure: D. Küppers, None; L. Tsiklauri, None; M. Hülser, None; K. Frommer, None; S. Rehart, None; C. Ospelt, None; U. Müller-Ladner, Biogen, 8; E. Neumann, None.

To cite this abstract in AMA style:

Küppers D, Tsiklauri L, Hülser M, Frommer K, Rehart S, Ospelt C, Müller-Ladner U, Neumann E. lncRNA H19 and Micro RNA 675-3p Are Altered by Visfatin During Osteogenesis [abstract]. Arthritis Rheumatol. 2020; 72 (suppl 10). https://acrabstracts.org/abstract/lncrna-h19-and-micro-rna-675-3p-are-altered-by-visfatin-during-osteogenesis/. Accessed .
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