TNIK对干扰素信号传导及内皮细胞对病毒感染反应的调控
TNIK regulation of interferon signaling and endothelial cell response to virus infection.
作者信息
Chau Khanh M, Dominic Abishai, Davis Eleanor L, Kotla Sivareddy, Berrios Estefani Turcios, Fahim Arsany, Arunesh Ashwin, Li Shengyu, Zhao Dongyu, Chen Kaifu, Davis Alan R, Nguyen Minh T H, Wang Yongxing, Evans Scott E, Wang Guangyu, Cooke John P, Abe Jun-Ichi, Huston David P, Le Nhat-Tu
机构信息
Department of Cardiovascular Sciences, Center for Cardiovascular Sciences, Academic Institute, Houston Methodist Research Institute, Weill Cornell Medical College, Houston, TX, United States.
Department of Molecular and Cellular Medicine, College of Medicine Texas A&M University, College Station, TX, United States.
出版信息
Front Cardiovasc Med. 2024 Jan 9;10:1213428. doi: 10.3389/fcvm.2023.1213428. eCollection 2023.
BACKGROUND
Traf2 and Nck-interacting kinase (TNIK) is known for its regulatory role in various processes within cancer cells. However, its role within endothelial cells (ECs) has remained relatively unexplored.
METHODS
Leveraging RNA-seq data and Ingenuity Pathway Analysis (IPA), we probed the potential impact of TNIK depletion on ECs.
RESULTS
Examination of RNA-seq data uncovered more than 450 Differentially Expressed Genes (DEGs) in TNIK-depleted ECs, displaying a fold change exceeding 2 with a false discovery rate (FDR) below 0.05. IPA analysis unveiled that TNIK depletion leads to the inhibition of the interferon (IFN) pathway [-log (-value) >11], downregulation of IFN-related genes, and inhibition of Hypercytokinemia/Hyperchemokinemia [-log (-value) >8]. The validation process encompassed qRT-PCR to evaluate mRNA expression of crucial IFN-related genes, immunoblotting to gauge STAT1 and STAT2 protein levels, and ELISA for the quantification of IFN and cytokine secretion in siTNIK-depleted ECs. These assessments consistently revealed substantial reductions upon TNIK depletion. When transducing HUVECs with replication incompetent E1-E4 deleted adenovirus expressing green fluorescent protein (Ad-GFP), it was demonstrated that TNIK depletion did not affect the uptake of Ad-GFP. Nonetheless, TNIK depletion induced cytopathic effects (CPE) in ECs transduced with wild-type human adenovirus serotype 5 (Ad-WT).
SUMMARY
Our findings suggest that TNIK plays a crucial role in regulating the EC response to virus infections through modulation of the IFN pathway.
背景
肿瘤坏死因子受体相关因子2与Nck相互作用激酶（TNIK）因其在癌细胞内各种过程中的调节作用而闻名。然而，其在内皮细胞（ECs）中的作用仍相对未被探索。
方法
利用RNA测序数据和 Ingenuity 通路分析（IPA），我们探究了TNIK缺失对内皮细胞的潜在影响。
结果
对RNA测序数据的检查发现，在TNIK缺失的内皮细胞中有超过450个差异表达基因（DEGs），其变化倍数超过2，错误发现率（FDR）低于0.05。IPA分析表明，TNIK缺失导致干扰素（IFN）通路受到抑制[-log（-值）>11]，IFN相关基因下调，以及高细胞因子血症/高趋化因子血症受到抑制[-log（-值）>8]。验证过程包括qRT-PCR评估关键IFN相关基因的mRNA表达、免疫印迹检测STAT1和STAT2蛋白水平，以及ELISA定量检测siTNIK缺失的内皮细胞中IFN和细胞因子的分泌。这些评估一致显示，TNIK缺失后有显著降低。在用表达绿色荧光蛋白的复制缺陷型E1-E4缺失腺病毒（Ad-GFP）转导人脐静脉内皮细胞（HUVECs）时，结果表明TNIK缺失不影响Ad-GFP的摄取。然而，TNIK缺失在用野生型人类5型腺病毒（Ad-WT）转导的内皮细胞中诱导了细胞病变效应（CPE）。
总结
我们的研究结果表明，TNIK通过调节IFN通路在调节内皮细胞对病毒感染的反应中起关键作用。
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