Epidemiology and Biostatistics, Department of Public Health, University of Southern Denmark, 5000, Odense C, Denmark.
Clinical Cell Biology, Pathology Research Unit, Department of Clinical Research, University of Southern Denmark, J. B. Winsløvs Vej 25, 1st Floor, 5000, Odense C, Denmark.
Clin Epigenetics. 2023 Mar 13;15(1):42. doi: 10.1186/s13148-023-01449-1.
Clinical trials have shown zoledronic acid as a potent bisphosphonate in preventing bone loss, but with varying potency between patients. Human osteoclasts ex vivo reportedly displayed a variable sensitivity to zoledronic acid > 200-fold, determined by the half-maximal inhibitory concentration (IC50), with cigarette smoking as one of the reported contributors to this variation. To reveal the molecular basis of the smoking-mediated variation on treatment sensitivity, we performed a DNA methylome profiling on whole blood cells from 34 healthy female blood donors. Multiple regression models were fitted to associate DNA methylation with ex vivo determined IC50 values, smoking, and their interaction adjusting for age and cell compositions.
We identified 59 CpGs displaying genome-wide significance (p < 1e-08) with a false discovery rate (FDR) < 0.05 for the smoking-dependent association with IC50. Among them, 3 CpGs have p < 1e-08 and FDR < 2e-03. By comparing with genome-wide association studies, 15 significant CpGs were locally enriched (within < 50,000 bp) by SNPs associated with bone and body size measures. Furthermore, through a replication analysis using data from a published multi-omics association study on bone mineral density (BMD), we could validate that 29 out of the 59 CpGs were in close vicinity of genomic sites significantly associated with BMD. Gene Ontology (GO) analysis on genes linked to the 59 CpGs displaying smoking-dependent association with IC50, detected 18 significant GO terms including cation:cation antiporter activity, extracellular matrix conferring tensile strength, ligand-gated ion channel activity, etc. CONCLUSIONS: Our results suggest that smoking mediates individual sensitivity to zoledronic acid treatment through epigenetic regulation. Our novel findings could have important clinical implications since DNA methylation analysis may enable personalized zoledronic acid treatment.
临床试验表明唑来膦酸作为一种有效的双膦酸盐，可预防骨质流失，但患者之间的疗效存在差异。据报道，人类破骨细胞对唑来膦酸的敏感性存在差异，其半数最大抑制浓度（IC50）差异超过 200 倍，而吸烟是导致这种差异的因素之一。为了揭示吸烟对治疗敏感性影响的分子基础，我们对 34 名健康女性献血者的全血细胞进行了 DNA 甲基化组谱分析。我们拟合了多元回归模型，将 DNA 甲基化与体外测定的 IC50 值、吸烟以及它们的相互作用与年龄和细胞组成进行关联。
我们发现 59 个 CpG 位点与 IC50 的吸烟依赖性关联具有全基因组意义（p < 1e-08，假发现率（FDR）< 0.05）。其中 3 个 CpG 位点的 p 值< 1e-08，FDR < 2e-03。通过与全基因组关联研究进行比较，15 个显著的 CpG 位点在与骨骼和身体大小测量相关的 SNPs 附近存在局部富集（< 50,000 bp 内）。此外，通过使用已发表的骨密度（BMD）多组学关联研究的数据进行复制分析，我们可以验证 59 个 CpG 位点中有 29 个与 BMD 显著相关的基因组位点非常接近。对与 IC50 的吸烟依赖性关联相关的 59 个 CpG 位点所连接的基因进行基因本体论（GO）分析，检测到 18 个显著的 GO 术语，包括阳离子：阳离子反向转运活性、细胞外基质赋予拉伸强度、配体门控离子通道活性等。
我们的研究结果表明，吸烟通过表观遗传调控介导个体对唑来膦酸治疗的敏感性。我们的新发现可能具有重要的临床意义，因为 DNA 甲基化分析可能实现唑来膦酸个体化治疗。
