Proteome profiling in IL-1β and VEGF-activated human umbilical vein endothelial cells delineates the interlink between inflammation and angiogenesis.
Proteome profiling in IL-1β and VEGF-activated human umbilical vein endothelial cells delineates the interlink between inflammation and angiogenesis.
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DOI：
10.1371/journal.pone.0179065
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发表时间：
2017
期刊：
影响因子：
3.7
通讯作者：
Gerner C
中科院分区：
文献类型：
--
作者：
Mohr T;Haudek-Prinz V;Slany A;Grillari J;Micksche M;Gerner C
内皮细胞代表炎症和血管生成的主要效应子，这些过程驱动多种病理状态，如动脉粥样硬化和癌症。炎症和血管生成两者在许多促炎蛋白具有促血管生成特性的意义上彼此相互关联，反之亦然。为了进一步阐明这种相互作用，我们提出了一个比较蛋白质组研究炎症和血管生成激活内皮细胞。分别用IL-1-β和VEGF刺激HUVEC。将培养的原代细胞分级分离成分泌的、细胞质和核蛋白级分，并进行处理以用于随后的LC-MS/MS分析。过滤获得的蛋白质谱中的组分特异性蛋白质，以解决潜在的交叉组分污染，进行比较计算生物学分析（GO-Term富集分析，加权基因共表达分析），并与已发表的IL-1β和VEGF刺激的HUVEC的mRNA谱进行比较。GO术语富集分析和比较途径分析揭示了诸如炎症激活的HUVEC的NOD和NfkB信号传导以及VEGF激活的HUVEC的VEGF和ErB信号传导的特征，其具有通过map激酶MAP 2K 2的潜在串扰。加权蛋白共表达网络分析揭示了迄今为止与炎症或血管生成中的驱动功能无关的几个潜在的枢纽基因，如HSPG 2、ANXA 3和GPI。“经典”炎症或血管生成标志物如IL 6、CXCL 8或CST 1在共表达网络内的较不中心的位置被发现。总之，本研究报告了一个框架的计算生物学为基础的蛋白质组学数据分析适用于细胞质，细胞核和细胞外部分的静止，炎症和血管生成激活HUVEC。与这些过程相关的新的潜在枢纽基因被成功地鉴定。
Endothelial cells represent major effectors in inflammation and angiogenesis, processes that drive a multitude of pathological states such as atherosclerosis and cancer. Both inflammation and angiogenesis are interconnected with each other in the sense that many pro-inflammatory proteins possess proangiogenic properties and vice versa. To elucidate this interplay further, we present a comparative proteome study of inflammatory and angiogenic activated endothelial cells. HUVEC were stimulated with interleukin 1-β and VEGF, respectively. Cultured primary cells were fractionated into secreted, cytoplasmic and nuclear protein fractions and processed for subsequent LC-MS/MS analysis. Obtained protein profiles were filtered for fraction-specific proteins to address potential cross fractional contamination, subjected to comparative computational biology analysis (GO-Term enrichment analysis, weighted gene co-expression analysis) and compared to published mRNA profiles of IL-1β respectively VEGF stimulated HUVEC. GO Term enrichment analysis and comparative pathway analysis revealed features such as NOD and NfkB signaling for inflammatory activated HUVEC and VEGF and ErB signaling for VEGF-activated HUVEC with potential crosstalk via map kinases MAP2K2. Weighted protein co-expression network analysis revealed several potential hub genes so far not associated with driver function in inflammation or angiogenesis such as HSPG2, ANXA3, and GPI. “Classical” inflammation or angiogenesis markers such as IL6, CXCL8 or CST1 were found in a less central position within the co-expression networks. In conclusion, this study reports a framework for the computational biology based analysis of proteomics data applied to cytoplasmic, nucleic and extracellular fractions of quiescent, inflammatory and angiogenic activated HUVEC. Novel potential hub genes relevant for these processes were successfully identified.