Session Title: Biology and Pathology of Bone and Joint - Poster I
Session Type: ACR Poster Session C
Session Time: 9:00AM-11:00AM
Background/Purpose: Osteoarthritis (OA) is a chronic and debilitating disease of the articulating joints in every population. Micro-RNAs are ~22 nucleotides long non-coding single stranded RNAs that mostly regulate post-transcriptional gene expression. In order to dissect the role of novel miRNAs in OA pathogenesis we exploited the next generation sequencing technology and comprehensively profiled the miRNAs expression in OA chondrocytes.
Methods: Chondrocytes were prepared by the enzymatic digestion of discarded human cartilage from donors who underwent total knee arthroplasty due to OA. Total RNA directly from the cartilage or from chondrocytes was prepared using Trizol and miRNeasy Mini kit respectively. RNA purity, integrity and NGS library integrity was verified using Agilent 2100 Bioanalyzer. The small RNA library was prepared using a kit (Illumina). Cluster generation and sequencing was performed on MiSeq system. The small RNA sequencing reads were aligned to genomic reference (hg19) and novel miRNA sequences were extracted from NGS data using Strand NGS software. miRNA targets were identified using open web bioinformatics program Diana Tools. Expression of mature miRNAs was quantified using individual miScript Primer Assay. Expression of KiSS1 and other OA related catabolic and anabolic genes was quantified using TaqMan Assays and of proteins by Western blotting. Chondrocytes were transfected with siRNAs, control miRNA, miR-N105 mimic or inhibitor using Amaxa Nucleofactor System.
To cite this abstract in AMA style:Shahidul Makki M, Haqqi T. KiSS1 Is a Regulator of ADAMTS4 and ADAMTS5 Expression and Is Post-Transcriptionally Regulated By Micro-RNA N105 in Human OA Chondrocytes [abstract]. Arthritis Rheumatol. 2016; 68 (suppl 10). https://acrabstracts.org/abstract/kiss1-is-a-regulator-of-adamts4-and-adamts5-expression-and-is-post-transcriptionally-regulated-by-micro-rna-n105-in-human-oa-chondrocytes/. Accessed August 4, 2020.
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ACR Meeting Abstracts - https://acrabstracts.org/abstract/kiss1-is-a-regulator-of-adamts4-and-adamts5-expression-and-is-post-transcriptionally-regulated-by-micro-rna-n105-in-human-oa-chondrocytes/