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.

National Institute of Health (NIH)
National Institute of Arthritis and Musculoskeletal and Skin Diseases (NIAMS)
Research Project (R01)
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Arthritis, Connective Tissue and Skin Study Section (ACTS)
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Mao, Su-Yau
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Northeast Ohio Medical University
Anatomy/Cell Biology
Schools of Medicine
United States
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Ansari, Mohammad Y; Khan, Nazir M; Ahmad, Nashrah et al. (2018) Genetic inactivation of ZCCHC6 suppresses IL-6 expression and reduces the severity of experimental osteoarthritis in mice. Arthritis Rheumatol :
Khan, Nazir M; Ahmad, Imran; Haqqi, Tariq M (2018) Nrf2/ARE pathway attenuates oxidative and apoptotic response in human osteoarthritis chondrocytes by activating ERK1/2/ELK1-P70S6K-P90RSK signaling axis. Free Radic Biol Med 116:159-171
Khan, Nazir M; Haqqi, Tariq M (2018) Epigenetics in osteoarthritis: Potential of HDAC inhibitors as therapeutics. Pharmacol Res 128:73-79
Ansari, M Y; Khan, N M; Ahmad, I et al. (2018) Parkin clearance of dysfunctional mitochondria regulates ROS levels and increases survival of human chondrocytes. Osteoarthritis Cartilage 26:1087-1097
Haseeb, Abdul; Makki, Mohammad Shahidul; Khan, Nazir M et al. (2017) Deep sequencing and analyses of miRNAs, isomiRs and miRNA induced silencing complex (miRISC)-associated miRNome in primary human chondrocytes. Sci Rep 7:15178
Makki, Mohammad Shahidul; Akhtar, Nahid; Haqqi, Tariq M (2017) An effective and efficient method of transfecting primary human chondrocytes in suspension. Anal Biochem 526:29-32
Akhtar, Nahid; Khan, Nazir M; Ashruf, Omer S et al. (2017) Inhibition of cartilage degradation and suppression of PGE2 and MMPs expression by pomegranate fruit extract in a model of posttraumatic osteoarthritis. Nutrition 33:1-13
Haseeb, Abdul; Ansari, Mohammad Yunus; Haqqi, Tariq M (2017) Harpagoside suppresses IL-6 expression in primary human osteoarthritis chondrocytes. J Orthop Res 35:311-320
Haseeb, Abdul; Khan, Nazir M; Ashruf, Omer S et al. (2017) A Polyphenol-rich Pomegranate Fruit Extract Suppresses NF-?B and IL-6 Expression by Blocking the Activation of IKK? and NIK in Primary Human Chondrocytes. Phytother Res 31:778-782
Khan, Nazir M; Haseeb, Abdul; Ansari, Mohammad Y et al. (2017) Wogonin, a plant derived small molecule, exerts potent anti-inflammatory and chondroprotective effects through the activation of ROS/ERK/Nrf2 signaling pathways in human Osteoarthritis chondrocytes. Free Radic Biol Med 106:288-301

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