背景:散发性甲状旁腺腺瘤(PA)是甲状旁腺功能亢进的最常见原因,然而,其发病机制仍不完全清楚。
方法:手术切除的PA样本,与正常的甲状旁腺(PG)组织一起,在甲状腺全切除术中偶然解剖,使用10×基因组学ChromiumDroplet平台和CellRanger软件进行单细胞RNA测序分析。进行基因集变异分析以表征标志性通路基因特征,和单细胞调控网络推断和聚类被用来分析转录因子调控子。进行免疫组织化学和免疫荧光以验证PA组织的细胞成分。siRNA敲低和基因过表达,除了定量聚合酶链反应,蛋白质印迹和细胞增殖试验,进行功能调查。
结果:与PG细胞相比,PA细胞(PACs)中的基因转录普遍增加。这与组蛋白-赖氨酸N-甲基转移酶2A(KMT2A)的高表达有关。高KMT2A水平可能通过上调原癌基因CCND2促进PAC增殖,这是由转录因子信号转导和转录激活因子3(STAT3)和GATA结合蛋白3(GATA3)介导的。PA组织大量浸润髓系细胞,而成纤维细胞,PA组织中的内皮细胞和巨噬细胞相对于PG组织中的相应细胞通常富含促炎基因特征.
结论:我们揭示了先前未被重视的KMT2A-STAT3/GATA3-CCND2轴和慢性炎症参与PA的发病机制。这些发现强调了KMT2A抑制和抗炎策略的治疗前景。强调未来研究将这些分子见解转化为实际应用的必要性。
结论:单细胞RNA测序揭示了比较散发性甲状旁腺腺瘤(PAs)与正常甲状旁腺的转录组目录。PA细胞显示与KMT2A上调相关的基因表达普遍增加。KMT2A介导的STAT3和GATA3上调是通过细胞周期蛋白D2促进PA细胞增殖的关键。PAs表现出促炎微环境,提示慢性炎症在PA发病机制中的潜在作用。
BACKGROUND: Sporadic parathyroid adenoma (PA) is the most common cause of hyperparathyroidism, yet the mechanisms involved in its pathogenesis remain incompletely understood.
METHODS: Surgically removed PA samples, along with normal parathyroid gland (PG) tissues that were incidentally dissected during total thyroidectomy, were analysed using single-cell RNA-sequencing with the 10× Genomics Chromium Droplet platform and Cell Ranger software. Gene set variation analysis was conducted to characterise hallmark pathway gene signatures, and single-cell regulatory network inference and clustering were utilised to analyse transcription factor regulons. Immunohistochemistry and immunofluorescence were performed to validate cellular components of PA tissues. siRNA knockdown and gene overexpression, alongside quantitative polymerase chain reaction, Western blotting and cell proliferation assays, were conducted for functional investigations.
RESULTS: There was a pervasive increase in gene transcription in PA cells (PACs) compared with PG cells. This is associated with high expression of histone-lysine N-methyltransferase 2A (
KMT2A). High KMT2A levels potentially contribute to promoting PAC proliferation through upregulation of the proto-oncogene CCND2, which is mediated by the transcription factors signal transducer and activator of transcription 3 (STAT3) and GATA binding protein 3 (GATA3). PA tissues are heavily infiltrated with myeloid cells, while fibroblasts, endothelial cells and macrophages in PA tissues are commonly enriched with proinflammatory gene signatures relative to their counterparts in PG tissues.
CONCLUSIONS: We revealed the previously underappreciated involvement of the
KMT2A‒STAT3/GATA3‒CCND2 axis and chronic inflammation in the pathogenesis of PA. These findings underscore the therapeutic promise of
KMT2A inhibition and anti-inflammatory strategies, highlighting the need for future investigations to translate these molecular insights into practical applications.
CONCLUSIONS: Single-cell RNA-sequencing reveals a transcriptome catalogue comparing sporadic parathyroid adenomas (PAs) with normal parathyroid glands. PA cells show a pervasive increase in gene expression linked to
KMT2A upregulation.
KMT2A-mediated STAT3 and GATA3 upregulation is key to promoting PA cell proliferation via cyclin D2. PAs exhibit a proinflammatory microenvironment, suggesting a potential role of chronic inflammation in PA pathogenesis.