BACKGROUND Aldosterone-producing adenomas (APAs) are a common cause of primary aldosteronism that can lead to cardiovascular complications if left untreated. Machine learning-based bioinformatics approaches have emerged as powerful tools for identifying potential disease markers, gaining widespread recognition in biomedical research. We aimed to use machine learning to discover novel biomarkers of APAs to identify new pathophysiological mechanisms. METHODS We applied 2 machine learning algorithms to published RNA sequencing data to identify APA feature genes. Validation was performed using APA tissue samples, spatial transcriptomics, pig adrenal glands, and in vitro assays in a human adrenocortical cell line. RESULTS Machine learning identified ATP2A3 as a key feature gene in APA, and its upregulation in APAs compared with the adjacent cortex was confirmed by spatial transcriptomics. In human adrenocortical cells, angiotensin II treatment increased ATP2A3 gene expression 9.15-fold. Silencing ATP2A3 decreased basal CYP11B2 expression and aldosterone secretion by 3.51-fold and 1.46-fold, respectively, and by 1.77-fold and 1.94-fold under angiotensin II stimulation. Dietary sodium restriction in pigs significantly increased ATP2A3 mRNA and protein levels. Spatial transcriptomics showed that APA cells exhibited higher ATP2A3 gene expression compared with all other adrenal cell types. The suppressive effect of ATP2A3 silencing on CYP11B2 expression was further enhanced by Ca2+ inhibitors. CONCLUSIONS The ATP2A3 gene is highly expressed in APA and is a key regulator of CYP11B2 expression and aldosterone production.

中文翻译：

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原发性醛固酮增多症中的 ATP2A3：基于机器学习的发现和功能验证。


背景 醛固酮生成腺瘤 （APA） 是原发性醛固酮增多症的常见原因，如果不及时治疗，可导致心血管并发症。基于机器学习的生物信息学方法已成为识别潜在疾病标志物的强大工具，在生物医学研究中获得了广泛认可。我们旨在使用机器学习来发现 APA 的新生物标志物，以确定新的病理生理机制。方法 我们将 2 种机器学习算法应用于已发布的 RNA 测序数据，以识别 APA 特征基因。使用 APA 组织样本、空间转录组学、猪肾上腺和人肾上腺皮质细胞系中的体外测定进行验证。结果 机器学习确定 ATP2A3 是 APA 中的关键特征基因，与相邻皮层相比，其在 APAs 中的上调通过空间转录组学得到证实。在人肾上腺皮质细胞中，血管紧张素 II 治疗使 ATP2A3 基因表达增加了 9.15 倍。沉默 ATP2A3 使基础CYP11B2表达和醛固酮分泌分别降低了 3.51 倍和 1.46 倍，在血管紧张素 II 刺激下降低了 1.77 倍和 1.94 倍。猪日粮钠限制显著提高了 ATP2A3 mRNA 和蛋白质水平。空间转录组学显示，与所有其他肾上腺细胞类型相比，APA 细胞表现出更高的 ATP2A3 基因表达。Ca2 + 抑制剂进一步增强了 ATP2A3 沉默对 CYP11B2 表达的抑制作用。结论 ATP2A3 基因在 APA 中高表达，是 CYP11B2 表达和醛固酮产生的关键调节因子。