Jiangsu Province Key Laboratory of Oral Diseases, Nanjing Medical University, No.136, Hanzhong Road, Nanjing, 210029, Jiangsu Province, China.
Department of Oral and Maxillofacial Surgery, Affiliated Hospital of Stomatology, Nanjing Medical University, Nanjing, 210029, China.
Mol Med. 2021 Jul 22;27(1):81. doi: 10.1186/s10020-021-00341-z.
Fibrous dysplasia (FD) is a bone marrow stromal cell (BMSC) disease caused by activating mutations of guanine nucleotide-binding protein alpha-stimulating activity polypeptide (GNAS) and is characterized by increased proliferative activity and disrupted osteogenesis of BMSCs. However, the molecular mechanisms regulating the pathophysiologic features of BMSCs in FD remain unknown. This study aimed to identify and verify the roles of the CREB1-miR-181a-5p regulatory loop in FD pathophysiology.
MicroRNA (miRNA) sequencing analysis was used to identify the possible miRNAs implicated in FD. The proliferation, apoptosis, and osteogenic differentiation of BMSCs, as well as the osteoclast-induced phenotype, were measured and compared after exogenous miR-181a-5p transfection into FD BMSCs or miR-181a-5p inhibitor transfection into normal BMSCs. Chromatin immunoprecipitation and luciferase reporter assays were performed to verify the interactions between CREB1 and miR-181a-5p and their effects on the FD pathological phenotype.
Compared to normal BMSCs, FD BMSCs showed decreased miR-181a-5p levels and exhibited increased proliferative activity, decreased apoptotic capacity, and impaired osteogenesis. FD BMSCs also showed a stronger osteoclast activation effect. miR-181a-5p overexpression reversed the pathophysiologic features of FD BMSCs, whereas miR-181a-5p suppression induced an FD-like phenotype in normal BMSCs. Mechanistically, miR-181a-5p was the downstream target of CREB1, and CREB1 was posttranscriptionally regulated by miR-181a-5p.
Our study identifies that the interaction loop between CREB1 and miR-181a-5p plays a crucial role in regulating the pathophysiologic features of FD BMSCs. MiR-181a-5p may be a potential therapeutic target for the treatment of FD.
纤维发育不良（FD）是一种由鸟嘌呤核苷酸结合蛋白α刺激活性多肽（GNAS）的激活突变引起的骨髓基质细胞（BMSC）疾病，其特征是 BMSC 的增殖活性增加和成骨作用受到破坏。然而，调节 FD 中 BMSC 病理生理特征的分子机制尚不清楚。本研究旨在鉴定和验证 CREB1-miR-181a-5p 调节环在 FD 病理生理学中的作用。
采用 microRNA（miRNA）测序分析鉴定可能与 FD 相关的 miRNAs。在 FD BMSCs 中转染外源性 miR-181a-5p 或在正常 BMSCs 中转染 miR-181a-5p 抑制剂后，测量和比较 BMSC 的增殖、凋亡和成骨分化以及破骨细胞诱导的表型。进行染色质免疫沉淀和荧光素酶报告基因测定以验证 CREB1 和 miR-181a-5p 之间的相互作用及其对 FD 病理表型的影响。
与正常 BMSCs 相比，FD BMSCs 显示 miR-181a-5p 水平降低，表现出增殖活性增加、凋亡能力降低和成骨作用受损。FD BMSCs 还表现出更强的破骨细胞激活作用。miR-181a-5p 的过表达逆转了 FD BMSCs 的病理生理特征，而 miR-181a-5p 的抑制在正常 BMSCs 中诱导出 FD 样表型。机制上，miR-181a-5p 是 CREB1 的下游靶标，而 CREB1 是由 miR-181a-5p 进行转录后调节的。
本研究确定 CREB1 和 miR-181a-5p 之间的相互作用环在调节 FD BMSC 的病理生理特征中起着关键作用。miR-181a-5p 可能是治疗 FD 的潜在治疗靶点。
