Atorvastatin and blood flow regulate expression of distinctive sets of genes in mouse carotid artery endothelium

Kumar, Sandeep; Sur, Sandeepa; Perez, Julian I.; Demos, Catherine; Kang, Dong‐Won; Kim, Chan Woo; Hu, Sarah; Xu, Ke; Yang, Jing; Jo, Hanjoong

doi:10.1016/bs.ctm.2021.08.004

Cited by 4 publications

(9 citation statements)

References 69 publications

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“…Thus, elucidating the potential relationship between ferroptosis and IH may provide new ideas and targets for the in-depth study of IH. Kumar et al mainly investigated the effects of atorvastatin at two time points on global endothelial gene expression by performing microarray studies using their mouse partial carotid ligation model ( Kumar and Sur, 2021 ). In our study, there were four time points in the dataset; however, a comprehensive analysis found no intersection across these, and FRGs were very few at 12 and 24 h, which not enough to support the following functional enrichment and pathway analysis.…”

Section: Discussionmentioning

confidence: 99%

Expression profiles and functions of ferroptosis-related genes in intimal hyperplasia induced by carotid artery ligation in mice

Zhang

Shen

et al. 2022

Front. Genet.

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Intimal hyperplasia (IH) is a prominent pathological event that occurs during in-stent restenosis and atherosclerosis. Ferroptosis, characterized by iron-dependent and lipid peroxidation, has become the recent focus of studies on the occurrence and progress of cardiovascular diseases. However, there are few studies on ferroptosis and IH. Therefore, we aimed to identify and validate ferroptosis-related markers in IH to explore new possibilities for IH diagnosis and treatment. The IH microarray dataset (GSE182291) was downloaded from the Gene Expression Omnibus (GEO) database and ferroptosis-related genes (FRGs) were obtained from the FerrDb databases. The differentially expressed genes (DEGs) were analyzed using the GEO2R. Overlapping was performed to identify the ferroptosis-related DEGs among the DEGs and FRGs. Then, clustering, Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment, and protein–protein interaction (PPI) analyses were performed. Subsequently, the hub genes were identified using Cytoscape and hub gene–transcription factors and hub gene–microRNA networks were constructed. Finally, real-time qPCR (RT-qPCR) and immunohistochemistry (IHC) were used to verify the mRNA and protein levels of the hub FRGs in IH. Thirty-four FRGs showing significantly different expression were identified from a total of 1,197 DEGs 2 days after ligation; 31 FRGs were selected from a total of 1,556 DEGs 14 days after ligation. The GO and KEGG analyses revealed that these 34 ferroptosis-related DEGs identified 2 days after ligation were mainly enriched in the basolateral plasma membrane, ferroptosis, lipid and atherosclerosis, and IL-17 signaling pathways. The 31 ferroptosis-related DEGs in endometrial hyperplasia identified 14 days after ligation were mainly enriched in response to oxidative stress, ferroptosis, tumor necrosis factor signaling pathway, and lipid and atherosclerosis. Five hub FRGs (Il1b, Ptgs2, Cybb, Cd44, and Tfrc) were identified using PPI networks; four hub FRGs (Il1b, Ptgs2, Cybb, and Cd44) were validated to be upregulated 2 and 14 days after ligation using RT-qPCR and show significantly different expression 14 days after ligation via IHC. Our findings verify the expression of hub DEGs related to ferroptosis in IH and elucidate the potential relationship between ferroptosis and IH, providing more evidence about the vital role of ferroptosis in IH.

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Section: Discussionmentioning

confidence: 99%

Expression profiles and functions of ferroptosis-related genes in intimal hyperplasia induced by carotid artery ligation in mice

Zhang

Shen

et al. 2022

Front. Genet.

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“…To identify FSTFs in mouse artery ECs in vivo, we first re-analyzed our genome-wide transcriptome data using the ECenriched bulk RNAs from the mouse PCL study (12). The gene array dataset (GSE182291) was comprised of transcript expression results of LCAs (exposed to d-flow) and RCAs (contralateral s-flow) obtained at four-time points: 12 h, 24 h, 48 h, and 2 weeks post-PCL.…”

Section: Identification and Validation Of Novel Factors Regulated In ...mentioning

confidence: 99%

“…D-flow and s-flow are recognized by mechanosensors, which trigger a wide spectrum of different endothelial responses. While s-flow/ULS promotes a healthy endothelial phenotype, d-flow/OSS induces atherosclerosis development and progression (7)(8)(9)(10)(11) by regulating gene expression on a genome-wide scale (9,(12)(13)(14)(15)(16)(17). Transcription factors regulated in a flow-sensitive manner (FSTFs) play a key role in genomewide gene expression by driving transcription of flow-sensitive genes (9,10,13,15,18).…”

Section: Introductionmentioning

confidence: 99%

“…To identify novel FSTFs in artery ECs in vivo, we reanalyzed two independent datasets that we previously reported using our mouse partial carotid ligation (PCL) model, the d-flow-induced mouse model of atherosclerosis (12,13). In the PCL model (Figure 1), three of the four caudal branches (left external carotid, internal carotid, and occipital artery) of the left common carotid artery (LCA) are ligated, leaving the superior thyroid artery patent.…”

Section: Introductionmentioning

confidence: 99%

“…The PCL causes d-flow with characteristic OSS in the LCA, rapidly inducing robust atherosclerosis within 2 weeks, directly demonstrating a causal relationship between d-flow and atherosclerosis (31,32). To understand the mechanisms of flow-induced atherosclerosis, we previously carried out a gene array study using endothelial-enriched "bulk" RNAs from the LCAs and RCAs obtained at four-time points (12 h, 24 h, 48 h, and 2 weeks) after the PCL surgery (12). In addition, we performed a single-cell RNA sequencing (scRNAseq) and single-cell Assay for Transposase-Accessible Chromatin with high-throughput sequencing (scATACseq) using the LCA and RCA RNAs obtained at 2-day and 2-week post-PCL time points (13).…”

Section: Introductionmentioning

confidence: 99%

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Sox13 is a novel flow-sensitive transcription factor that prevents inflammation by repressing chemokine expression in endothelial cells

Demos

Johnson

Andueza

et al. 2022

Front. Cardiovasc. Med.

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Atherosclerosis is a chronic inflammatory disease and occurs preferentially in arterial regions exposed to disturbed blood flow (d-flow) while the stable flow (s-flow) regions are spared. D-flow induces endothelial inflammation and atherosclerosis by regulating endothelial gene expression partly through the flow-sensitive transcription factors (FSTFs). Most FSTFs, including the well-known Kruppel-like factors KLF2 and KLF4, have been identified from in vitro studies using cultured endothelial cells (ECs). Since many flow-sensitive genes and pathways are lost or dysregulated in ECs during culture, we hypothesized that many important FSTFs in ECs in vivo have not been identified. We tested the hypothesis by analyzing our recent gene array and single-cell RNA sequencing (scRNAseq) and chromatin accessibility sequencing (scATACseq) datasets generated using the mouse partial carotid ligation model. From the analyses, we identified 30 FSTFs, including the expected KLF2/4 and novel FSTFs. They were further validated in mouse arteries in vivo and cultured human aortic ECs (HAECs). These results revealed 8 FSTFs, SOX4, SOX13, SIX2, ZBTB46, CEBPβ, NFIL3, KLF2, and KLF4, that are conserved in mice and humans in vivo and in vitro. We selected SOX13 for further studies because of its robust flow-sensitive regulation, preferential expression in ECs, and unknown flow-dependent function. We found that siRNA-mediated knockdown of SOX13 increased endothelial inflammatory responses even under the unidirectional laminar shear stress (ULS, mimicking s-flow) condition. To understand the underlying mechanisms, we conducted an RNAseq study in HAECs treated with SOX13 siRNA under shear conditions (ULS vs. oscillatory shear mimicking d-flow). We found 94 downregulated and 40 upregulated genes that changed in a shear- and SOX13-dependent manner. Several cytokines, including CXCL10 and CCL5, were the most strongly upregulated genes in HAECs treated with SOX13 siRNA. The robust induction of CXCL10 and CCL5 was further validated by qPCR and ELISA in HAECs. Moreover, the treatment of HAECs with Met-CCL5, a specific CCL5 receptor antagonist, prevented the endothelial inflammation responses induced by siSOX13. In addition, SOX13 overexpression prevented the endothelial inflammation responses. In summary, SOX13 is a novel conserved FSTF, which represses the expression of pro-inflammatory chemokines in ECs under s-flow. Reduction of endothelial SOX13 triggers chemokine expression and inflammatory responses, a major proatherogenic pathway.

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Restoration of normal blood flow in atherosclerotic arteries promotes plaque stabilization

et al. 2023

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Atorvastatin and blood flow regulate expression of distinctive sets of genes in mouse carotid artery endothelium

Cited by 4 publications

References 69 publications

Expression profiles and functions of ferroptosis-related genes in intimal hyperplasia induced by carotid artery ligation in mice

Expression profiles and functions of ferroptosis-related genes in intimal hyperplasia induced by carotid artery ligation in mice

Sox13 is a novel flow-sensitive transcription factor that prevents inflammation by repressing chemokine expression in endothelial cells

Restoration of normal blood flow in atherosclerotic arteries promotes plaque stabilization

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