Campus for Ageing and Vitality, Biosciences Institute, Newcastle University, Newcastle-upon-Tyne, United Kingdom.
Regenerative Medicine, Stem Cells, Transplantation, Biosciences Institute, Newcastle University, Newcastle upon Tyne, United Kingdom.
Elife. 2024 Oct 14;12:RP89701. doi: 10.7554/eLife.89701.
Changes in chondrocyte gene expression can contribute to the development of osteoarthritis (OA), and so recognition of the regulative processes during chondrogenesis can lead to a better understanding of OA. microRNAs (miRNAs) are key regulators of gene expression in chondrocytes/OA, and we have used a combined experimental, bioinformatic, and systems biology approach to explore the multiple miRNA-mRNA interactions that regulate chondrogenesis. A longitudinal chondrogenesis bioinformatic analysis identified paralogues miR-199a-5p and miR-199b-5p as pro-chondrogenic regulators. Experimental work in human cells demonstrated alteration of miR-199a-5p or miR-199b-5p expression led to significant inverse modulation of key chondrogenic genes and extracellular matrix production. miR-199a/b-5p targets and were identified by inhibition experiments and verified as direct targets by luciferase assay. The experimental work was used to generate and parameterise a multi-miRNA 14-day chondrogenesis kinetic model to be used as a repository for the experimental work and as a resource for further investigation of this system. This is the first multi-miRNA model of a chondrogenesis-based system, and highlights the complex relationships between regulatory miRNAs, and their target mRNAs.
软骨细胞基因表达的变化可能导致骨关节炎（OA）的发生，因此，对软骨发生过程中的调节过程的认识可以更好地理解 OA。microRNAs（miRNAs）是软骨细胞/OA 中基因表达的关键调节剂，我们采用了综合的实验、生物信息学和系统生物学方法来探索调节软骨发生的多种 miRNA-mRNA 相互作用。一项纵向软骨发生生物信息学分析确定了 miRNA-199a-5p 和 miRNA-199b-5p 的同源物作为促软骨形成的调节剂。在人类细胞中的实验工作表明，miR-199a-5p 或 miR-199b-5p 的表达改变导致关键软骨形成基因和细胞外基质产生的显著反向调节。通过抑制实验鉴定了 miR-199a/b-5p 的靶标 和 ，并通过荧光素酶测定验证为直接靶标。该实验工作用于生成和参数化一个多 miRNA 14 天软骨发生动力学模型，用作实验工作的存储库，并作为进一步研究该系统的资源。这是第一个基于软骨发生的系统的多 miRNA 模型，突出了调节 miRNA 与其靶 mRNA 之间的复杂关系。
