CCN5 Reduces Ligamentum Flavum Hypertrophy by Modulating the TGF-β Pathway

Sunghyeok Ye; Woo-Keun Kwon; Taegeun Bae; Sunghyun Kim; Jang-Bo Lee; Tai-Hyoung Cho; Jung-Yul Park; Kyoungmi Kim; Junho K Hur; Junseok W Hur

doi:10.1002/jor.24425

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CCN5 Reduces Ligamentum Flavum Hypertrophy by Modulating the TGF-β Pathway

Journal article

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CCN5 Reduces Ligamentum Flavum Hypertrophy by Modulating the TGF-β Pathway

Sunghyeok Ye, Woo-Keun Kwon, Taegeun Bae, Sunghyun Kim, Jang-Bo Lee, Tai-Hyoung Cho, Jung-Yul Park, Kyoungmi Kim, Junho K Hur and Junseok W Hur

Journal of orthopaedic research, Vol.37(12), pp.2634-2644

12/2019

DOI: 10.1002/jor.24425

PMCID: PMC6899892

PMID: 31334871

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Abstract

Ligamentum flavum hypertrophy (LFH) is the most important component of lumbar spinal canal stenosis. Although the pathophysiology of LFH has been extensively studied, no method has been proposed to prevent or treat it. Since the transforming growth factor-β (TGF-β) pathway is known to be critical in LFH pathology, we investigated whether LFH could be prevented by blocking or modulating the TGF-β mechanism. Human LF cells were used for the experiments. First, we created TGF-β receptor 1 (TGFBR1) knock out (KO) cells with CRISPR (clustered regularly interspaced short palindromic repeats)/Cas9 biotechnology and treated them with TGF-β1 to determine the effects of blocking the TGF-β pathway. Subsequently, we studied the effect of CCN5, which has recently been proposed to modulate the TGF-β pathway. To assess the predisposition toward fibrosis, α-smooth muscle actin (αSMA), fibronectin, collagen-1, collagen-3, and CCN2 were evaluated with quantitative real-time polymerase chain reaction, western blotting, and immunocytochemistry. The TGFBR1 KO LF cells were successfully constructed with high KO efficiency. In wild-type (WT) cells, treatment with TGF-β1 resulted in the overexpression of the messenger RNA (mRNA) of fibrosis-related factors. However, in KO cells, the responses to TGF-β1 stimulation were significantly lower. In addition, CCN5 and TGF-β1 co-treatment caused a notable reduction in mRNA expression levels compared with TGF-β1 stimulation only. The αSMA protein expression increased with TGF-β1 but decreased with CCN5 treatment. TGF-β1 induced LF cell transdifferentiation from fibroblasts to myofibroblasts. However, this cell transition dramatically decreased in the presence of CCN5. In conclusion, CCN5 could prevent LFH by modulating the TGF-β pathway. © 2019 The Authors. Journal of Orthopaedic Research published by Wiley Periodicals, Inc. on behalf of Orthopaedic Research Society. J Orthop Res 37:2634-2644, 2019.

Actins - analysis

CCN Intercellular Signaling Proteins - pharmacology

Cell Transdifferentiation - drug effects

Cells, Cultured

Clustered Regularly Interspaced Short Palindromic Repeats

Fibroblasts - pathology

Fibrosis

Humans

Hypertrophy

Ligamentum Flavum - drug effects

Ligamentum Flavum - pathology

Myofibroblasts - pathology

Receptor, Transforming Growth Factor-beta Type I - physiology

Repressor Proteins - pharmacology

Signal Transduction - physiology

Transforming Growth Factor beta - physiology

Details

Title: Subtitle: CCN5 Reduces Ligamentum Flavum Hypertrophy by Modulating the TGF-β Pathway
Creators: Sunghyeok Ye - RnD center, GeneCker, Seoul, Korea
Woo-Keun Kwon - Korea University
Taegeun Bae - Kyung Hee University
Sunghyun Kim - Department of Molecular and Computational Biology, University of Southern California, Los Angeles, California
Jang-Bo Lee - Korea University
Tai-Hyoung Cho - Korea University
Jung-Yul Park - Department of Neurosurgery, College of Medicine, Korea University Anam Hospital, 73, Inchon-ro, Seongbuk-gu, Seoul, 02841, Korea
Kyoungmi Kim - Korea University
Junho K Hur - Kyung Hee University
Junseok W Hur - Korea University
Resource Type: Journal article
Publication Details: Journal of orthopaedic research, Vol.37(12), pp.2634-2644
DOI: 10.1002/jor.24425
PMID: 31334871
PMCID: PMC6899892
NLM abbreviation: J Orthop Res
ISSN: 0736-0266
eISSN: 1554-527X
Grant note: NRF-2018M3A9H3021707 / Basic Science Research Program through the National Research Foundation (NRF) funded by the Korean Ministry of Education, Science and Technology (KR) NRF-2019R1C1C1010602 / Basic Science Research Program through the National Research Foundation (NRF) funded by the Korean Ministry of Education, Science and Technology (KR) K1808641 / College of Medicine, Korea University NRF-2017R1E1A1A01074529 / Basic Science Research Program through the National Research Foundation (NRF) funded by the Korean Ministry of Education, Science and Technology (KR) K1722461 / College of Medicine, Korea University K1809751 / College of Medicine, Korea University NRF-2017R1D1A1B03035760 / Basic Science Research Program through the National Research Foundation (NRF) funded by the Korean Ministry of Education, Science and Technology (KR) NRF-2017R1D1A1B03035094 / Basic Science Research Program through the National Research Foundation (NRF) funded by the Korean Ministry of Education, Science and Technology (KR)
Language: English
Date published: 12/2019
Academic Unit: Neurosurgery
Record Identifier: 9984460330402771

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