AB0074 KRÜPPEL-LIKE FACTOR 10 IS A IMPORTANT MODULATORY FACTOR OF CHONDROCYTE HYPERTROPHY IN DEVELOPING SKELETON. (2nd June 2020)

• Record Type: Journal Article
• Authors: Ko, J. Y.; Lee, E.; Kim, J.; Im, G. I.
• Is Part Of: Annals of the rheumatic diseases. Volume 79(2020)Supplement 1
• Page Start: 1337
• Page End: 1337
• Publication Date: 2020-06-02
• Subjects: Rheumatism -- Periodicals; 616.723005
• DOI: 10.1136/annrheumdis-2020-eular.2065
• Languages: English
• ISSNs: 0003-4967

Abstract

Abstract : Background: To investigate the functional role of KLF10 as a modulator of chondrocyte hypertrophy in developmental skeleton, the developmental characteristics in the long bone of KLF10 knockout mice and characteristics of MSCs from KLF10 KO mice were investigated regarding chondrogenesis and osteogenesis. Delayed long bone growth and delayed formation of primary ossification center were observed in an early embryonic stage in KLF10 KO mouse along with very low Ihh expression in epiphyseal plate. While the chondrogenic potential of mMSCs appeared normal or slight decrerased in KLF10 KO mice, osteogenesis and hypertrophy were extensively suppressed. KLF10 was found to be a mediator of chondrocyte hypertrophy in developing skeleton. Suppression of KLF10 may be considered as a new strategy for preventing hypertrophy in cartilage regeneration using MSCs. Objectives: Investigated the functional role of KLF10 to present new insights into the transcriptional network regulating skeletal development and provide a novel strategy for preventing aberrant hypertrophic differentiation in cartilage regeneration strategies using MSCs. Methods: Generation of KLF10 KO mice and genotyping / Skeletal preparations, embryo heights, and mineralized bone length measurements / Histological and Fluorescent Immunohistochemical Analysis / ALP staining and activity / Alizarin red staining / Von Kossa staining and calcium salts quantification / Isolation and Establishment of Mouse Clonal MSC Lines / Chondrogenic pellet culture and differentiation of mMSCs / DNA Quantification and GAG Contents Analysis / Rq-PCR Analysis / Statistics Results: The overall results showed that mMSCs from KLF10 KO mice have significantly decreased osteogenic potential with very low Ihh expression while an increase in chondrogenic potential was not significant. In addition to Ihh promotor demonstrated in our previous study, KLF10 can activate Runx2 expression through its proximal-promoter region. Thus, KLF10 may indirectly stimulate Ihh expression upstream of Runx2 or directly bind to Ihh promoter and activate Ihh expression. As shown in this and out previous study, KLF10 also enhances Wnt/β-catenin signalling in MSCs. KLF10 modulates β-catenin sub-cellular localization and enhances Wnt signalling in osteoblasts. Conclusion: In conclusion, primary ossification in KLF10 KO mice was critically delayed during early endochondral bone development. KLF10 KO inhibited hypertrophy via reduced Ihh expression in developing skeleton. TGF-β-induced hypertrophy was inhibited during chondrogenesis of KLF10 KO mMSCs. Our findings present new insights into the transcriptional-network system of skeletal development and provide a novel strategy for suppressing hypertrophy in cartilage tissue engineering. Acknowledgments: This research was supported by the National Research Foundation of Korea (NRF-2019R1H1A2039685 and 2019R1I1A1A01043778). Disclosure of Interests: None declared