Zhuang Xingxing, Xiao Fei, Chen Feihu, Ni Shoudong
Department of Pharmacy, Chaohu Hospital of Anhui Medical University, No. 64 North Chaohu Road, Chaohu, Anhui, 238000, People's Republic of China.
School of Pharmacy, Anhui Medical University, No. 81 Meishan Road, Hefei, Anhui, 230000, People's Republic of China.
Clin Exp Nephrol. 2025 Jan 21. doi: 10.1007/s10157-024-02620-5.
This study seeks to investigate the fundamental molecular processes through which histone deacetylase 9 (HDAC9) governs the proliferation of glomerular mesangial cells in the context of immunoglobulin A nephropathy (IgAN) and to identify novel targets for clinical research on IgAN.
Data from high-throughput RNA sequencing for IgAN were procured from the Gene Expression Omnibus database to assess the expression profiles and clinical diagnostic significance of histone deacetylase family proteins (HDACs). Blood samples from 20 IgAN patients were employed in RT-qPCR analysis, and the spearman linear regression method was utilized to analyze the clinical correlation. The proliferation of glomerular mesangial cells (GMCs) under the influence of HDAC9 was examined using the 5-ethynyl-2'-deoxyuridine (EdU) assay. Proteins interacting with HDAC9 were predicted utilizing the STRING database. Immunoprecipitation and protein immunoblotting employing anti-acetylated lysine antibodies were conducted to determine the acetylation status of calmodulin-like protein 6 (CALML6).
Analysis of the GSE141295 dataset revealed a significant upregulation of HDAC9 expression in IgAN and the results of RT-qPCR demonstrated a substantial increase in HDAC9 expression in IgAN patients. Receiver operating characteristic (ROC) analysis indicated that the area under the curve (AUC) value for HDAC9 were 0.845 and Spearman correlation analysis showed that HDAC9 expression was positively correlated with blood levels of blood urea nitrogen (BUN) and serum creatinine (Crea). The EdU cell proliferation assay indicated that HDAC9 facilitated the excessive proliferation of GMCs. The STRING database and recovery experiments identified CALML6 as a downstream effector of HDAC9 in controlling abnormal GMC multiplication. Co-immunoprecipitation assays demonstrated that HDAC9 modulates CALML6 expression through acetylation modification.
HDAC9 is markedly upregulated in IgAN, and it mediates the excessive proliferation of GMCs by regulating the deacetylation of CALML6.
本研究旨在探讨组蛋白去乙酰化酶9（HDAC9）在免疫球蛋白A肾病（IgAN）背景下调控肾小球系膜细胞增殖的基本分子过程，并确定IgAN临床研究的新靶点。
从基因表达综合数据库获取IgAN的高通量RNA测序数据，以评估组蛋白去乙酰化酶家族蛋白（HDACs）的表达谱和临床诊断意义。采用20例IgAN患者的血样进行RT-qPCR分析，并利用Spearman线性回归方法分析临床相关性。使用5-乙炔基-2'-脱氧尿苷（EdU）检测法检测HDAC9影响下肾小球系膜细胞（GMCs）的增殖情况。利用STRING数据库预测与HDAC9相互作用的蛋白质。进行免疫沉淀和使用抗乙酰化赖氨酸抗体的蛋白质免疫印迹，以确定钙调蛋白样蛋白6（CALML6）的乙酰化状态。
对GSE141295数据集的分析显示，IgAN中HDAC9表达显著上调，RT-qPCR结果表明IgAN患者中HDAC9表达大幅增加。受试者操作特征（ROC）分析表明，HDAC9的曲线下面积（AUC）值为0.845，Spearman相关性分析显示HDAC9表达与血尿素氮（BUN）和血清肌酐（Crea）水平呈正相关。EdU细胞增殖检测表明HDAC9促进了GMCs的过度增殖。STRING数据库和回收实验确定CALML6是HDAC9控制GMC异常增殖的下游效应分子。免疫共沉淀实验表明HDAC9通过乙酰化修饰调节CALML6表达。
HDAC9在IgAN中明显上调，它通过调节CALML6的去乙酰化介导GMCs的过度增殖。
