The choroid within the human eye contains a rich milieu of cells including melanocytes. Human choroidal melanocytes (HCMs) absorb light, regulate free radical production, and were recently shown to modulate inflammation. This study aimed to identify key genes and pathways involved in the inflammatory response of HCMs through the use of RNA-seq. Primary HCMs were cultured from donor choroids, RNA was extracted from control and lipopolysaccharide (LPS)-treated HCMs, and mRNA was sequenced. Functional annotation and pathway analysis were performed using gene ontology and gene set enrichment analyses. Representative RNA-seq results were verified with RT-qPCR and protein measurements. We detected 100 differentially expressed genes including an array of CCL and CXCL cytokines and mediators of cell–cell and cell–matrix adhesion, such as ICAM1, CLDN1, CCN3, ITGA1 and ITGA11. Functional annotation showed that these gene sets control inflammatory pathways, immune cell trafficking, cell–cell adhesion, interactions with the extracellular matrix and blood vessels, angiogenesis and epithelial-to-mesenchymal transitions. Our study provides insights into the transcriptional regulation of primary HCMs in response to inflammatory stimuli and identifies novel melanocyte-driven mechanisms potentially involved in choroidal homeostasis and inflammation.

中文翻译：

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黑素细胞在人类脉络膜微环境和炎症中的作用：来自转录组的见解

人眼内的脉络膜包含丰富的细胞环境，包括黑素细胞。人类脉络膜黑色素细胞 (HCM) 吸收光，调节自由基的产生，并且最近被证明可以调节炎症。本研究旨在通过使用 RNA-seq 确定参与 HCM 炎症反应的关键基因和途径。从供体脉络膜培养原代 HCM，从对照和脂多糖 (LPS) 处理的 HCM 中提取 RNA，并对 mRNA 进行测序。使用基因本体和基因集富集分析进行功能注释和通路分析。通过 RT-qPCR 和蛋白质测量验证了代表性的 RNA-seq 结果。我们检测到 100 个差异表达基因，包括一系列 CCL 和 CXCL 细胞因子以及细胞-细胞和细胞-基质粘附的介质，例如ICAM1、CLDN1、CCN3、ITGA1和ITGA11。功能注释显示这些基因组控制炎症通路、免疫细胞运输、细胞-细胞粘附、与细胞外基质和血管的相互作用、血管生成和上皮-间质转化。我们的研究提供了对原发性 HCM 响应炎症刺激的转录调控的见解，并确定了可能参与脉络膜稳态和炎症的新型黑素细胞驱动机制。