AUTHOR=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 

TITLE=TNIK regulation of interferon signaling and endothelial cell response to virus infection

JOURNAL=Frontiers in Cardiovascular Medicine

VOLUME=10

YEAR=2024

URL=https://www.frontiersin.org/journals/cardiovascular-medicine/articles/10.3389/fcvm.2023.1213428

DOI=10.3389/fcvm.2023.1213428

ISSN=2297-055X

ABSTRACT=<sec><title>Background</title><p>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.</p></sec><sec><title>Methods</title><p>Leveraging RNA-seq data and Ingenuity Pathway Analysis (IPA), we probed the potential impact of TNIK depletion on ECs.</p></sec><sec><title>Results</title><p>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 (<italic>p</italic>-value) &gt;11], downregulation of IFN-related genes, and inhibition of Hypercytokinemia/Hyperchemokinemia [-log (<italic>p</italic>-value) &gt;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).</p></sec><sec><title>Summary</title><p>Our findings suggest that TNIK plays a crucial role in regulating the EC response to virus infections through modulation of the IFN pathway.</p></sec>