Li Yong, Yue Yang, Cen Chaoqun, Liu Bo
Department of Emergency, The Third Xiangya Hospital, Central South University, Changsha, 410013, Hunan, China.
Department of Orthopedics, The Third Xiangya Hospital, Central South University, Changsha, 410013, Hunan, China.
Hum Cell. 2025 Aug 27;38(5):152. doi: 10.1007/s13577-025-01273-0.
miR-487a-3p shows the potential of modulating adipose-derived stem cells (ADSCs) differentiation. This study aimed to investigate the mechanism of miR-487a-3p on the osteogenic differentiation of ADSCs. In this work, ADSCs were induced to differentiate into osteoblasts. miR-487a-3p were regulated by miRNA mimics or inhibitors in ADSCs. Wnt family member 5A (WNT5A) siRNA was used to reverse miR-487a-3p inhibitor-induced effects on WNT5A expression in ADSCs. Fat mass and obesity-associated protein (FTO) in ADSCs were altered by shRNAs or overexpression vectors. Calcium nodule, ALP activity, and biomarkers of osteogenic differentiation (OD) were investigated. Rats received ovariectomy (OVX) to construct osteoporosis (OP) model, followed by ADSCs transplantation. Histopathological changes, bone histomorphometry, and detection of OD biomarkers were performed. We found that osteogenesis induced a decrease in miR-487a-3p expression and an increase in FTO expression. miR-487a-3p upregulation inhibited the OD of ADSCs, including decreases in calcium nodule formation, ALP activity, and OD biomarkers. miR-487a-3p downregulation showed the opposite role in OD. miR-487a-3p negatively regulated WNT5A in ADSCs. WNT5A silence reversed the effect of miR-487a-3p downregulation on OD. FTO silence caused the increase in m6A of pri-miR-487a. FTO overexpression inhibited DGCR8 recruitment in pri-miR-487a, and reversed the effect of miR-487a-3p upregulation in OD. ADSCs transplantation improved OP symptom in rats, including improvement of femur tissue, increase in percent bone volume and trabecular number, and upregulation in OD biomarkers. miR-487a-3p downregulation enhanced the therapeutic role of ADSC in rats with OP. Collectively, FTO regulated pri-miR-487a maturation via m6A-dependent pathway, which altered the WNT5A-mediated osteogenesis of ADSCs.
miR-487a-3p具有调控脂肪来源干细胞（ADSCs）分化的潜力。本研究旨在探究miR-487a-3p对ADSCs成骨分化的作用机制。在本研究中，诱导ADSCs分化为成骨细胞。通过miRNA模拟物或抑制剂调控ADSCs中miR-487a-3p的表达。使用Wnt家族成员5A（WNT5A）的小干扰RNA（siRNA）来逆转miR-487a-3p抑制剂对ADSCs中WNT5A表达的影响。通过短发夹RNA（shRNAs）或过表达载体改变ADSCs中脂肪量和肥胖相关蛋白（FTO）的表达。检测钙结节、碱性磷酸酶（ALP）活性和成骨分化（OD）的生物标志物。对大鼠进行卵巢切除术（OVX）以构建骨质疏松症（OP）模型，随后进行ADSCs移植。进行组织病理学变化、骨组织形态计量学分析以及OD生物标志物检测。我们发现成骨过程中miR-487a-3p表达降低，FTO表达升高。上调miR-487a-3p抑制了ADSCs的OD，包括钙结节形成减少、ALP活性降低以及OD生物标志物水平下降。下调miR-487a-3p在OD中发挥相反作用。miR-487a-3p在ADSCs中负向调控WNT5A。沉默WNT5A可逆转下调miR-487a-3p对OD的影响。沉默FTO导致pri-miR-487a的N6-甲基腺苷（m6A）增加。过表达FTO抑制pri-miR-487a中DGCR8的募集，并逆转上调miR-487a-3p对OD的影响。ADSCs移植改善了大鼠的OP症状，包括股骨组织改善、骨体积百分比和小梁数量增加以及OD生物标志物上调。下调miR-487a-3p增强了ADSCs对OP大鼠的治疗作用。总体而言，FTO通过m6A依赖途径调控pri-miR-487a的成熟，从而改变WNT5A介导的ADSCs成骨作用。
