Abstract：
BACKGROUND: Shuangshen granules (SSG), a nationally patented Chinese medicinal formula, including Panax quinquefolium L., Panax notoginseng (Burkill) F. H. Chen, and Cordyceps sinensis (Berk.) Sacc., has demonstrated remarkable therapeutic effects on pancreatic cancer in clinical treatment for nearly 10 years. Previous pharmacological researches have found that its main components, including ginsenosides and cordycepin have anticancer or preventive effects on pancreatic ductal adenocarcinoma (PDAC), which may be associated with immune metabolism. However, the underlying pharmacological mechanism of SSG in the truncation effect of PDAC progression is still unclear.
OBJECTIVE: To comprehensively understand the infiltrating immune cells during the different stages of the PDAC development chain and search for immune-related biomarkers that could potentially serve as drug targets through bioinformatic analysis. Meanwhile, the truncation effect of SSG on PDAC progression was also investigated.
METHODS: The gene expression profiles at different PDAC developmental stages, including normal pancreas, pancreatic intraepithelial neoplasia (PanIN), and PDAC, were retrieved from the GEO database. The GEO2R tool was used to identify differentially expressed genes among the three groups. Functional enrichment analysis was performed with the GSEA software and Metascape platform. The CIBERSORT algorithm evaluated immune cell infiltration in the three groups, and immune-related biomarkers were identified. Correlation analysis was employed to examine the association between immune cells and the biomarkers. One of these biomarkers was selected for immunohistochemistry validation in human samples. Lastly, the effectiveness of SSG against PDAC progression and the influence on the selected biomarker were validated in vivo. The underlying pharmacological mechanisms were also explored.
RESULTS: One dataset was obtained, where the functional enrichment of DEGs primarily involved immune effector processes and cytokine production of immune cells. The differential immune cells reflected during the progression from PanIN to PDAC were B memory cells, monocytes, M2 macrophages, and activated dendritic cells. The upregulation of ACTA2 was closely associated with M2 macrophage regulation. The immunohistochemistry on human samples validated significant differences in ACTA2 expression levels as the PDAC progressed. Moreover, animal experiments revealed that the national patented drug SSG ameliorated the pathological changes, decreased the expression of ACTA2 and its functional protein α-smooth muscle actin during PDAC progression. The underlying pharmacological mechanism was related to the regulation of macrophage polarization and downregulation of TGF-β/Smad signaling pathway.
CONCLUSIONS: The immunosuppressive environment changes during the PDAC progression. ACTA2 is a potential immuned-target for drug prevention of PDAC, while SSG could be a promising drug candidate.
OBJECTIVE: To comprehensively understand the infiltrating immune cells during the different stages of the PDAC development chain and search for immune-related biomarkers that could potentially serve as drug targets through bioinformatic analysis. Meanwhile, the truncation effect of SSG on PDAC progression was also investigated.
METHODS: The gene expression profiles at different PDAC developmental stages, including normal pancreas, pancreatic intraepithelial neoplasia (PanIN), and PDAC, were retrieved from the GEO database. The GEO2R tool was used to identify differentially expressed genes among the three groups. Functional enrichment analysis was performed with the GSEA software and Metascape platform. The CIBERSORT algorithm evaluated immune cell infiltration in the three groups, and immune-related biomarkers were identified. Correlation analysis was employed to examine the association between immune cells and the biomarkers. One of these biomarkers was selected for immunohistochemistry validation in human samples. Lastly, the effectiveness of SSG against PDAC progression and the influence on the selected biomarker were validated in vivo. The underlying pharmacological mechanisms were also explored.
RESULTS: One dataset was obtained, where the functional enrichment of DEGs primarily involved immune effector processes and cytokine production of immune cells. The differential immune cells reflected during the progression from PanIN to PDAC were B memory cells, monocytes, M2 macrophages, and activated dendritic cells. The upregulation of ACTA2 was closely associated with M2 macrophage regulation. The immunohistochemistry on human samples validated significant differences in ACTA2 expression levels as the PDAC progressed. Moreover, animal experiments revealed that the national patented drug SSG ameliorated the pathological changes, decreased the expression of ACTA2 and its functional protein α-smooth muscle actin during PDAC progression. The underlying pharmacological mechanism was related to the regulation of macrophage polarization and downregulation of TGF-β/Smad signaling pathway.
CONCLUSIONS: The immunosuppressive environment changes during the PDAC progression. ACTA2 is a potential immuned-target for drug prevention of PDAC, while SSG could be a promising drug candidate.
摘要：
背景：双参颗粒（SSG），国家专利的中药配方，包括西洋参，三七（伯基尔）F.H.Chen，和冬虫夏草（Berk。)萨克。,在近10年的临床治疗中对胰腺癌有显著的治疗效果。以往的药理研究发现,其主要成分,包括人参皂苷和虫草素对胰腺导管腺癌（PDAC）具有抗癌或预防作用，这可能与免疫代谢有关。然而,SSG在PDAC进展的截断效应中的潜在药理机制尚不清楚。
目的：全面了解PDAC发育链不同阶段的浸润免疫细胞，并通过生物信息学分析寻找可能作为药物靶标的免疫相关生物标志物。同时,还研究了SSG对PDAC进展的截断效应.
方法：不同PDAC发育阶段的基因表达谱，包括正常的胰腺,胰腺上皮内瘤变（PanIN），PDAC,是从GEO数据库中检索的。GEO2R工具用于鉴定三组之间的差异表达基因。用GSEA软件和Metascape平台进行功能富集分析。TheCIBERSORT算法评估了三组的免疫细胞浸润，并鉴定了免疫相关的生物标志物。采用相关性分析来检查免疫细胞与生物标志物之间的关联。选择这些生物标志物之一用于人样品中的免疫组织化学验证。最后,在体内验证了SSG对抗PDAC进展的有效性以及对所选生物标志物的影响.还探讨了潜在的药理机制。
结果：获得了一个数据集，其中DEGs的功能富集主要涉及免疫效应子过程和免疫细胞的细胞因子产生。从PanIN到PDAC的过程中反映的差异免疫细胞是B记忆细胞，单核细胞,M2巨噬细胞，和激活的树突状细胞。ACTA2的上调与M2巨噬细胞的调控密切相关。人样品的免疫组织化学证实了随着PDAC进展ACTA2表达水平的显著差异。此外，动物实验表明，国家专利药物SSG改善了病理变化，PDAC过程中ACTA2及其功能蛋白α-平滑肌肌动蛋白的表达降低。潜在的药理机制与调节巨噬细胞极化和下调TGF-β/Smad信号通路有关。
结论：在PDAC进展过程中，免疫抑制环境发生变化。ACTA2是PDAC药物预防的潜在免疫靶标，而SSG可能是一个有前途的候选药物。
目的：全面了解PDAC发育链不同阶段的浸润免疫细胞，并通过生物信息学分析寻找可能作为药物靶标的免疫相关生物标志物。同时,还研究了SSG对PDAC进展的截断效应.
方法：不同PDAC发育阶段的基因表达谱，包括正常的胰腺,胰腺上皮内瘤变（PanIN），PDAC,是从GEO数据库中检索的。GEO2R工具用于鉴定三组之间的差异表达基因。用GSEA软件和Metascape平台进行功能富集分析。TheCIBERSORT算法评估了三组的免疫细胞浸润，并鉴定了免疫相关的生物标志物。采用相关性分析来检查免疫细胞与生物标志物之间的关联。选择这些生物标志物之一用于人样品中的免疫组织化学验证。最后,在体内验证了SSG对抗PDAC进展的有效性以及对所选生物标志物的影响.还探讨了潜在的药理机制。
结果：获得了一个数据集，其中DEGs的功能富集主要涉及免疫效应子过程和免疫细胞的细胞因子产生。从PanIN到PDAC的过程中反映的差异免疫细胞是B记忆细胞，单核细胞,M2巨噬细胞，和激活的树突状细胞。ACTA2的上调与M2巨噬细胞的调控密切相关。人样品的免疫组织化学证实了随着PDAC进展ACTA2表达水平的显著差异。此外，动物实验表明，国家专利药物SSG改善了病理变化，PDAC过程中ACTA2及其功能蛋白α-平滑肌肌动蛋白的表达降低。潜在的药理机制与调节巨噬细胞极化和下调TGF-β/Smad信号通路有关。
结论：在PDAC进展过程中，免疫抑制环境发生变化。ACTA2是PDAC药物预防的潜在免疫靶标，而SSG可能是一个有前途的候选药物。
