Feasibility study of m6A modifier genes YTHDF2 and FMR1 for assessing the risks of postmenopausal osteoporosis
Objective To assess the expression characteristics of m6A-modified genes in postmenopausal osteoporosis(PMOP)based on microarrays,single-cell RNA sequencing(scRNA-seq)and clinical cohorts.Methods Menopausal females who underwent bone densitometry during March 2023-March 2024 in our hospital were selected as study subjects.After balancing age,years of menopause and BMI variables according to the propensity score matching method,they were enrolled in the group on a 1:1 basis,and finally 86 cases were included in each of the PMOP and control group.The m6A modification gene expression characteristics and distribution were explored based on microarrays GSE13850,GSE56815,GSE56814 and scRNA-seq dataset GSE147287.Four m6A modifier genes,IGFBP3,YTHDF2,FMR1 and RBM15,were verified for expression characteristics in neutrophils using RT-qPCR and protein immunoblotting.Expression profiles in plasma YTHDF2 and RBM15 were detected and assessed using ELISA.Plasma YTHDF2 and RBM15 expressions were assessed for correlation with bone mineral density(BMD)by Pearson test phase analysis.The accuracy of plasma YTHDF2 and RBM15 in diagnosing PMOP was assessed by receiver operating characteristic(ROC)curves.Results Microarray analysis showed that IGFBP3 expression was up-regulated in PMOP(P<0.05);YTHDF2,FMR1 and RBM15 expressions were down-regulated in PMOP(P<0.05);YTHDF2 was expressed in a variety of cells,but mainly in neutrophils and osteoblasts;RBM15 was mainly expressed in neutrophils;IGFBP3 and FMR1 were expressed in small amounts in all 9 cells.RT-qPCR and protein immunoblotting results showed that the expressions of both YTHDF2 and RBM15 were reduced in PMOP neutrophils(P<0.05).Compared with the control group,plasma YTHDF2 and RBM15 expressions were significantly reduced in PMOP(P<0.05).YTHDF2 showed positive correlations with LS_BMD(r=0.306)and FN_BMD(r=0.258)(P<0.05);RBM15 showed positive correlations with LS_BMD(r=0.551)and FN_BMD(r=0.426)(P<0.05).The area under the curve(AUC)of plasma YTHDF2 and RBM15 for the diagnosis of PMOP were 0.913(0.869-0.957)and 0.896(0.846-0.946),respectively.Conclusions The m6A-modified genes YTHDF2 and FMR1 have reduced expressions in PMOP,and the assessment and characterisation of YTHDF2 and FMR1 expressions can provide new insights for PMOP risk assessment and treatment.
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