The precise patterning of the developing skeletal framework relies on the appropriate control of chondrogenesis, a multistep process during which mesenchymal cells differentiate into chondrocytes. Articular cartilage destruction is a central factor in the pathogenesis of joint diseases, including osteoarthritis (OA) and rheumatoid arthritis (RA). Research on the molecular mechanisms of chondrogenesis is of developmental biological interest and has the potential to lead to novel approaches in tissue engineering and novel arthritis treatments. MicroRNAs (miRNAs) are a family of ~22-nucleotide (nt) noncoding RNAs that regulate gene expression by posttranscriptional mechanisms. To date, chondrocyte specific miRNAs, their upstream molecular signals, and target genes are largely unknown. In our preliminary experiments, we performed microarray profiling of miRNA expression in chondrocytes and miR-140 expression in cartilage development during embryogenesis and adult cartilage chondrocytes. These studies demonstrated that miR-140 has the most cartilage specific expression pattern. Cartilage specific Sox9 null mice did not express miR-140 and the potential Sox9 binding region close to pre-miR-140 was associated with Sox9 by ChIP assay and EMSA, suggesting that Sox9 is a key regulator of miR-140 gene expression. miR-140 expression was down-regulated in osteoarthritic cartilage compared with normal cartilage and by IL-1? stimulation of chondrocytes. miR-140 null mice showed earlier onset of osteoarthritic changes in knee joint at 6 months. Microarray and bioinformatics analysis revealed a set of possible miR-140 target genes in chondrocytes, including ADAMTS-5, which is a key tissue-degrading enzyme in osteoarthritis. These observations support the hypothesis that miR-140 is a novel regulator of cartilage development and homeostasis and those changes in its expression and function play an important role in diseases affecting articular cartilage, such as osteoarthritis. We propose the following specific aims:
Aim 1 : Investigate the transcriptional regulation of miR-140 expression by Sox9 in mesenchymal stem cells and chondrocytes.
Aim 2 : Examine the role of miR-140 in chondrogenesis and cartilage development.
Aim 3 : Determine the expression and function of miR-140 in osteoarthritis.
Aim 4 : Identify novel target genes of miR-140 in chondrocytes and mesenchymal stem cells. The proposed studies have the potential to reveal important new regulatory pathways that control cartilage development and homeostasis and open new insight on disease mechanisms and therapeutic interventions.

Public Health Relevance

Despite substantial recent progress in understanding OA pathogenesis, disease-modifying therapies for the most prevalent joint disease are not available. Examining the role of miR-140 in MSC and chondrocytes may provide new insight into cartilage cell biology as well as a basis for novel approaches in tissue engineering and therapeutic strategies for osteoarthritis.

Agency
National Institute of Health (NIH)
Institute
National Institute of Arthritis and Musculoskeletal and Skin Diseases (NIAMS)
Type
Research Project (R01)
Project #
5R01AR050631-10
Application #
8660645
Study Section
Skeletal Biology Structure and Regeneration Study Section (SBSR)
Program Officer
Tyree, Bernadette
Project Start
2003-03-01
Project End
2015-05-31
Budget Start
2014-06-01
Budget End
2015-05-31
Support Year
10
Fiscal Year
2014
Total Cost
$401,980
Indirect Cost
$190,300
Name
Scripps Research Institute
Department
Type
DUNS #
781613492
City
La Jolla
State
CA
Country
United States
Zip Code
92037
Inui, Masafumi; Mokuda, Sho; Sato, Tempei et al. (2018) Dissecting the roles of miR-140 and its host gene. Nat Cell Biol 20:516-518
Matsuzaki, Tokio; Alvarez-Garcia, Oscar; Mokuda, Sho et al. (2018) FoxO transcription factors modulate autophagy and proteoglycan 4 in cartilage homeostasis and osteoarthritis. Sci Transl Med 10:
Mochizuki, Yusuke; Chiba, Tomoki; Kataoka, Kensuke et al. (2018) Combinatorial CRISPR/Cas9 Approach to Elucidate a Far-Upstream Enhancer Complex for Tissue-Specific Sox9 Expression. Dev Cell 46:794-806.e6
Kataoka, Kensuke; Matsushima, Takahide; Ito, Yoshiaki et al. (2018) Bhlha9 regulates apical ectodermal ridge formation during limb development. J Bone Miner Metab 36:64-72
Nakamichi, Ryo; Kataoka, Kensuke; Asahara, Hiroshi (2018) Essential role of Mohawk for tenogenic tissue homeostasis including spinal disc and periodontal ligament. Mod Rheumatol 28:933-940
Otero, Miguel; Peng, Haibing; Hachem, Karim El et al. (2017) ELF3 modulates type II collagen gene (COL2A1) transcription in chondrocytes by inhibiting SOX9-CBP/p300-driven histone acetyltransferase activity. Connect Tissue Res 58:15-26
Ito, Yoshiaki; Inoue, Atsushi; Seers, Timothy et al. (2017) Identification of targets of tumor suppressor microRNA-34a using a reporter library system. Proc Natl Acad Sci U S A 114:3927-3932
Koda, Naoki; Sato, Tempei; Shinohara, Masahiro et al. (2017) The transcription factor mohawk homeobox regulates homeostasis of the periodontal ligament. Development 144:313-320
Nakasuji, Takashi; Ogonuki, Narumi; Chiba, Tomoki et al. (2017) Complementary Critical Functions of Zfy1 and Zfy2 in Mouse Spermatogenesis and Reproduction. PLoS Genet 13:e1006578
Miyata, Kohei; Naito, Masashi; Miyata, Tomoko et al. (2017) Bisulfite Sequencing for DNA Methylation Analysis of Primary Muscle Stem Cells. Methods Mol Biol 1668:3-13

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