Gerini Giulia, Traversa Alice, Cece Fabrizio, Cassandri Matteo, Pontecorvi Paola, Camero Simona, Nannini Giulia, Romano Enrico, Marampon Francesco, Venneri Mary Anna, Ceccarelli Simona, Angeloni Antonio, Amedei Amedeo, Marchese Cinzia, Megiorni Francesca
Department of Experimental Medicine, Sapienza University of Rome, 00161 Rome, Italy.
Department of Life Sciences, Health and Health Professions, Link Campus University, 00165 Rome, Italy.
Cells. 2025 Jan 17;14(2):135. doi: 10.3390/cells14020135.
Adipose-derived mesenchymal stem cells (ASCs) are commonly employed in clinical treatment for various diseases due to their ability to differentiate into multi-lineage and anti-inflammatory/immunomodulatory properties. Preclinical studies support their use for bone regeneration, healing, and the improvement of functional outcomes. However, a deeper understanding of the molecular mechanisms underlying ASC biology is crucial to identifying key regulatory pathways that influence differentiation and enhance regenerative potential. In this study, we employed the NanoString nCounter technology, an advanced multiplexed digital counting method of RNA molecules, to comprehensively characterize differentially expressed transcripts involved in metabolic pathways at distinct time points in osteogenically differentiating ASCs treated with or without the pan-DNMT inhibitor RG108. annotation and gene ontology analysis highlighted the activation of ethanol oxidation, ROS regulation, retinoic acid metabolism, and steroid hormone metabolism, as well as in the metabolism of lipids, amino acids, and nucleotides, and pinpointed potential new osteogenic drivers like AOX1 and ADH1A. RG108-treated cells, in addition to the upregulation of the osteogenesis-related markers RUNX2 and ALPL, showed statistically significant alterations in genes implicated in transcriptional control (MYCN, MYB, TP63, and IRF1), ethanol oxidation (ADH1C, ADH4, ADH6, and ADH7), and glucose metabolism (SLC2A3). These findings highlight the complex interplay of the metabolic, structural, and signaling pathways that orchestrate osteogenic differentiation. Furthermore, this study underscores the potential of epigenetic drugs like RG108 to enhance ASC properties, paving the way for more effective and personalized cell-based therapies for bone regeneration.
脂肪来源的间充质干细胞（ASCs）由于其能够分化为多谱系细胞以及具有抗炎/免疫调节特性，因而常用于各种疾病的临床治疗。临床前研究支持将其用于骨再生、愈合以及改善功能结局。然而，深入了解ASC生物学背后的分子机制对于确定影响分化并增强再生潜力的关键调控途径至关重要。在本研究中，我们采用了NanoString nCounter技术，这是一种先进的RNA分子多重数字计数方法，以全面表征在用或不用泛DNA甲基转移酶抑制剂RG108处理的成骨分化ASCs中，在不同时间点参与代谢途径的差异表达转录本。注释和基因本体分析突出了乙醇氧化、ROS调节、视黄酸代谢和类固醇激素代谢的激活，以及脂质、氨基酸和核苷酸的代谢，并确定了潜在的新的成骨驱动因子，如AOX1和ADH1A。除了成骨相关标志物RUNX2和ALPL的上调外，RG108处理的细胞在涉及转录控制（MYCN、MYB、TP63和IRF1）、乙醇氧化（ADH1C、ADH4、ADH6和ADH7）和葡萄糖代谢（SLC2A3）的基因中显示出统计学上的显著变化。这些发现突出了协调成骨分化的代谢、结构和信号通路之间的复杂相互作用。此外，本研究强调了像RG108这样的表观遗传药物增强ASC特性的潜力，为更有效和个性化的基于细胞的骨再生治疗铺平了道路。
