An estimated 27 million Americans age 25 and older have osteoarthritis (OA), a degenerative disease of articular cartilage characterized by loss of the cartilage matrix, mainly collagen and proteoglycans, leading to joint tissue destruction and loss of function. Before age 45, more men than women have osteoarthritis; after age 45, it is more common in women. The pro-inflammatory cytokine IL-6 has recently gained interest due to its elevated levels in synovial fluid and sera from OA patients and its capacity to upregulate the expression of MMP-13 and inhibit the expression of type-II collagen. Mature miRNAs are non-coding RNAs that inhibit the translation and stability of target mRNAs and are now known to undergo addition of non-template nucleotides (largely uridines) to the 3' ends (3'NTAs). The enzymes responsible for 3' end modifications of miRNAs have only recently been identified in cancer and immune cells but have not yet been described in cartilage or chondrocytes. Our preliminary data show that (1) ZCCHC6-a nucleotidyl transferase-is highly expressed in damaged OA cartilage; and (2) regulates IL-6 expression in human chondrocytes. ZCCHC6 is known to uridylate mRNAs thus suggesting that ZCCHC6-mediated miRNA 3'end modifications could play a significant role in the post-transcriptional regulation of IL-6, and possibly of other inflammatory mediators, in OA. We will pursue 4 specific aims to test the fundamental hypothesis that ZCCHC6 uridylates mature miRNAs in chondrocytes to regulate IL-6 expression and inflammation in OA.
Specific Aim -1. Studies will test the hypothesis that cytokine targeting miRNAs are uridylated in OA cartilage.
Specific Aim -2. We will test the hypothesis that miRNA uridylation abrogates repression of IL-6 expression in human OA chondrocytes.
Specific Aim -3. We will test the hypothesis that expression of ZCCHC6 is essential for IL-6 expression in human OA chondrocytes.
Specific Aim -4. In an animal model of human OA we will test the hypothesis that ZCCHC6 mediates miRNA uridylation and regulate IL-6 expression during the induction and progression of the disease. Knowledge gained from these studies will be important since by understanding how miRNA 3'end modifications affect the expression of inflammatory mediators in OA may lead to the development of novel biomarkers and therapies for the effective treatment of OA.

Public Health Relevance

Public Health Relevance Osteoarthritis is the most common form of arthritis and major cause of physical disability among the elderly population worldwide. Studies in this application are focused on understanding how the cytokine expression is dysregulated in chondrocytes and contributes to OA pathogenesis. Results of these studies may help in the design of therapeutic agents that target novel mechanisms involved in cartilage catabolism in OA.

Agency
National Institute of Health (NIH)
Institute
National Institute of Arthritis and Musculoskeletal and Skin Diseases (NIAMS)
Type
Research Project (R01)
Project #
5R01AR067056-03
Application #
9247878
Study Section
Arthritis, Connective Tissue and Skin Study Section (ACTS)
Program Officer
Mao, Su-Yau
Project Start
2015-04-01
Project End
2020-03-31
Budget Start
2017-04-01
Budget End
2018-03-31
Support Year
3
Fiscal Year
2017
Total Cost
$509,142
Indirect Cost
$173,296
Name
Northeast Ohio Medical University
Department
Anatomy/Cell Biology
Type
Schools of Medicine
DUNS #
077779882
City
Rootstown
State
OH
Country
United States
Zip Code
44272
Sondag, Gregory R; Haqqi, Tariq M (2016) The Role of MicroRNAs and Their Targets in Osteoarthritis. Curr Rheumatol Rep 18:56
Khan, Nazir M; Ansari, Mohammad Y; Haqqi, Tariq M (2016) Sucrose, But Not Glucose, Blocks IL1-β-Induced Inflammatory Response in Human Chondrocytes by Inducing Autophagy via AKT/mTOR Pathway. J Cell Biochem :
Haseeb, Abdul; Ansari, Mohammad Yunus; Haqqi, Tariq M (2016) Harpagoside suppresses IL-6 expression in primary human osteoarthritis chondrocytes. J Orthop Res :
Magrey, Marina N; Haqqi, Tariq; Haseeb, Abdul (2016) Identification of plasma microRNA expression profile in radiographic axial spondyloarthritis-a pilot study. Clin Rheumatol 35:1323-7
Makki, Mohammad S; Haqqi, Tariq M (2016) Histone deacetylase inhibitor vorinostat (SAHA, MK0683) perturb miR-9-MCPIP1 axis to block IL-1β-induced IL-6 expression in human OA chondrocytes. Connect Tissue Res :1-12
Ansari, Mohammad Y; Haqqi, Tariq M (2016) Interleukin-1β induced Stress Granules Sequester COX-2 mRNA and Regulates its Stability and Translation in Human OA Chondrocytes. Sci Rep 6:27611
Makki, Mohammad Shahidul; Haqqi, Tariq M (2015) miR-139 modulates MCPIP1/IL-6 expression and induces apoptosis in human OA chondrocytes. Exp Mol Med 47:e189
Makki, Mohammad S; Haseeb, Abdul; Haqqi, Tariq M (2015) MicroRNA-9 promotion of interleukin-6 expression by inhibiting monocyte chemoattractant protein-induced protein 1 expression in interleukin-1β-stimulated human chondrocytes. Arthritis Rheumatol 67:2117-28