慢性肾脏病成骨细胞的染色质可及性和表观遗传 DNA 修饰:对慢性肾脏病儿科患者骨骼和成骨细胞的研究

IF 3.4 Q2 ENDOCRINOLOGY & METABOLISM JBMR Plus Pub Date : 2024-01-04 DOI:10.1093/jbmrpl/ziad015
Aline Martin, R. Kawaguchi, Qing Wang, I. Salusky, Renata C Pereira, K. Wesseling-Perry
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引用次数: 0

摘要

成熟缺陷是 CKD 患者成骨细胞系细胞的固有特征。这些缺陷在体内外持续存在,表明 CKD 会诱导骨细胞发生表观遗传学变化。为了深入了解哪些信号通路有助于CKD介导的表观遗传学驱动的成骨细胞成熟障碍,我们通过RNA-Seq和甲基化史诗鉴定了9名患有终末期肾病的青少年和年轻成人透析患者以及3名健康参考者的原代成骨细胞的RNA表达和DNA甲基化模式。还对成骨细胞子集进行了 ATAC-Seq。从髂嵴提取骨基质蛋白表达,并通过蛋白质组学进行评估。GSEA 分析用于确定染色质可及性、DNA 甲基化和 RNA 表达模式发生一致改变的信号通路。利用 shRNA 抑制了原发性成骨细胞中的单个基因,并在体外鉴定了矿化特征。我们还使用 3-(4,5-二甲基噻唑-2-基)-5-(3-羧基甲氧基苯基)-2-(4-磺酸苯基)-2H-四氮唑(MTS)掺入法评估了 NFAT 信号抑制的效果。我们发现,在 CKD 成骨细胞中,对成骨细胞分化至关重要的信号通路被强烈下调。GSEA 发现了 NFAT 信号靶点的高度甲基化变化、染色质可及性差异和 RNA 表达改变。抑制 NFAT 可减少成骨细胞的增殖。对成骨细胞 RNA 表达和整个骨基质组成的综合分析确定了 13 对潜在的配体-受体。总之,CKD 成骨细胞的表观遗传学变化与多个成骨细胞基因和信号通路的表达改变有关。NFAT 信号的增加可能是 CKD 成骨细胞成熟受损的原因之一。表观遗传学变化还与骨基质的改变有关,这可能会导致骨质脆弱。有必要开展进一步研究,以阐明受这些基因改变影响的通路,因为阐明这些通路对于纠正 CKD 患者骨病的潜在生物学特性至关重要。
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Chromatin accessibility and epigenetic DNA modifications in CKD osteoblasts: a study of bone and osteoblasts from pediatric patients with chronic kidney disease
Maturation defects are intrinsic features of osteoblast lineage cells in CKD patients. These defects persist ex vivo, suggesting that CKD induces epigenetic changes in bone cells. To gain insights into which signaling pathways contribute to CKD-mediated, epigenetically-driven, impairments in osteoblast maturation, we characterized RNA expression and DNA methylation patterns by RNA-Seq and Methylation Epic in primary osteoblasts from 9 adolescent and young adult dialysis patients with end-stage kidney disease and 3 healthy references. ATAC-Seq was also performed on a subset of osteoblasts. Bone matrix protein expression was extracted from iliac crest and evaluated by proteomics. GSEA analysis was used to establish signaling pathways consistently altered in chromatin accessibility, DNA methylation, and RNA expression patterns. Single genes were suppressed in primary osteoblasts using shRNA and mineralization characterized in vitro. The effect of NFAT signaling suppression was also assessed using 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) incorporation. We found that signaling pathways critical for osteoblast differentiation were strongly downregulated in CKD osteoblasts. GSEA identified highly significant methylation changes, differential chromatin accessibility, and altered RNA expression in NFAT signaling targets. NFAT inhibition reduced osteoblast proliferation. Combined analysis of osteoblast RNA expression and whole bone matrix composition identified thirteen potential ligand-receptor pairs were identified. In summary, epigenetic changes in CKD osteoblasts associate with altered expression of multiple osteoblast genes and signaling pathways. An increase in NFAT signaling may play a role in impaired CKD osteoblast maturation. Epigenetic changes also associate with an altered bone matrix which may contribute to bone fragility. Further studies are necessary to elucidate the pathways affected by these genetic alterations since elucidating these pathways will be vital to correcting the underlying biology of bone disease in the CKD population.
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来源期刊
JBMR Plus
JBMR Plus Medicine-Orthopedics and Sports Medicine
CiteScore
5.80
自引率
2.60%
发文量
103
审稿时长
8 weeks
期刊最新文献
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