Gene targeting of the transcription factor Mohawk in rats causes heterotopic ossification of Achilles tendon via failed tenogenesis
| Autor: | Akio Kishida, Shizuko Ichinose, Ryo Nakamichi, Atsushi Okawa, Yoshiaki Ito, Masahiro Shinohara, Martin Lotz, Hiroshi Asahara, Tomohiro Kayama, Takuya Oyaizu, Kazuyoshi Yagishita, Satoshi Yamashita, Hidetsugu Suzuki, Naoki Koda |
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| Jazyk: | angličtina |
| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
musculoskeletal diseases Male SOX9 Biology Achilles Tendon 03 medical and health sciences Gene Knockout Techniques Tissue engineering Osteogenesis medicine Animals Rats Wistar Homeodomain Proteins Achilles tendon Multidisciplinary Adipogenesis Ossification Heterotopic Gene targeting Anatomy Biological Sciences Chondrogenesis musculoskeletal system Tendon Cell biology 030104 developmental biology medicine.anatomical_structure Knockout mouse Stress Mechanical |
| Popis: | Cell-based or pharmacological approaches for promoting tendon repair are currently not available because the molecular mechanisms of tendon development and healing are not well understood. Although analysis of knockout mice provides many critical insights, small animals such as mice have some limitations. In particular, precise physiological examination for mechanical load and the ability to obtain a sufficient number of primary tendon cells for molecular biology studies are challenging using mice. Here, we generated Mohawk (Mkx)(-/-) rats by using CRISPR/Cas9, which showed not only systemic hypoplasia of tendons similar to Mkx(-/-) mice, but also earlier heterotopic ossification of the Achilles tendon compared with Mkx(-/-) mice. Analysis of tendon-derived cells (TDCs) revealed that Mkx deficiency accelerated chondrogenic and osteogenic differentiation, whereas Mkx overexpression suppressed chondrogenic, osteogenic, and adipogenic differentiation. Furthermore, mechanical stretch stimulation of Mkx(-/-) TDCs led to chondrogenic differentiation, whereas the same stimulation in Mkx(+/+) TDCs led to formation of tenocytes. ChIP-seq of Mkx overexpressing TDCs revealed significant peaks in tenogenic-related genes, such as collagen type (Col)1a1 and Col3a1, and chondrogenic differentiation-related genes, such as SRY-box (Sox)5, Sox6, and Sox9 Our results demonstrate that Mkx has a dual role, including accelerating tendon differentiation and preventing chondrogenic/osteogenic differentiation. This molecular network of Mkx provides a basis for tendon physiology and tissue engineering. |
| Databáze: | OpenAIRE |
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