Serum microRNA signatures are indicative of skeletal fractures in post-menopausal women with and without type 2 diabetes and influence osteo-genic differentiation of mesenchymal stem cells

The objective of this study was the identification of serum microRNAs that can differentiate osteoporotic fracture patients with and without type-2 diabetes from healthy control subjects. For that purpose circulating microRNAs were profiled by real-time quantitative PCR using a custom 384-well panel in 200 µl serum samples. Univariate and multivariate statistical tools were used in order to identify single as well as combinations of circulating microRNas that were characteristic of patients with prevalent osteoporotic fractures: a qRT-PCR-based classifier consisting of miR-550a-5p, miR-96-5p, miR-32-3p and miR-486-5p can distinguish T2D women with (DMFx) and without fragility fractures (DM) with high specifitiy and sensitivity (AUC = 0.93). A classifier consisting of miR-188-3p, miR-382-3p, miR-942 and miR-155-5p was capable of differentiating between postmenopausal women with osteoporotic fractures and fracture-free controls with an AUC of 0.98. qPCR circulating microRNA profiling. Four groups were defined: postmenopausal women without type-2 diabetes and no history of osteoporotic fractures (CO); postmenopausal women without type-2 diabetes and a history of at least one (non-recent) osteoporotic fractures (FX); postmenopausal women with type-2 diabetes and no history of osteoporotic fractures (DM); postmenopausal women with type-2 diabetes and a history of at least one (non-recent) osteoporotic fractures (DMFX)

本研究旨在鉴定可区分合并与未合并2型糖尿病的骨质疏松性骨折患者，以及健康对照受试者的血清微RNA（microRNA）。为此，本研究采用定制384孔微RNA检测板的实时定量PCR（real-time quantitative PCR）技术，对200μL血清样本中的循环微RNA进行表达谱分析。本研究通过单变量及多变量统计工具，筛选可区分现患骨质疏松性骨折患者的单种循环微RNA及其组合。最终构建的基于qRT-PCR的分类器由miR-550a-5p、miR-96-5p、miR-32-3p及miR-486-5p组成，可高特异性与高灵敏度地区分合并脆性骨折的2型糖尿病女性（DMFx组）与未合并脆性骨折的2型糖尿病女性（DM组），受试者工作特征曲线下面积（Area Under the Curve，AUC）达0.93。另一款由miR-188-3p、miR-382-3p、miR-942及miR-155-5p组成的分类器，可区分合并骨质疏松性骨折的绝经后女性与无骨折健康对照，AUC达0.98。本研究涉及循环微RNA的qPCR表达谱分析。本研究共定义四组研究对象：①无2型糖尿病且无骨质疏松性骨折病史的绝经后女性（CO组）；②无2型糖尿病但有至少1次（非近期）骨质疏松性骨折病史的绝经后女性（FX组）；③有2型糖尿病但无骨质疏松性骨折病史的绝经后女性（DM组）；④有2型糖尿病且有至少1次（非近期）骨质疏松性骨折病史的绝经后女性（DMFX组）