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Effect of the sGC activator BAY 60-2770 on human proximal tubular (HK-2) cells under hypoxia and after TGF-β1 treatment

ABSTRACT: Effect of the sGC activator BAY 60-2770 on human proximal tubular (HK-2) cells under hypoxia and after TGF-β1 treatment

PROVIDER: PRJNA1121890 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR29337534_1.fastq.gz	Fastqsanger.gz
	SRR29337534_2.fastq.gz	Fastqsanger.gz
	SRR29337535_1.fastq.gz	Fastqsanger.gz
	SRR29337535_2.fastq.gz	Fastqsanger.gz
	SRR29337536_1.fastq.gz	Fastqsanger.gz

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Effect of the sGC activator BAY 60-2770 on human proximal tubular (HK-2) cells under hypoxia and after TGF-β1 treatment

Project description:Increasing cGMP levels by modulating soluble guanylyl cyclase (sGC) can be renoprotective. While sGC stimulators can modulate cGMP levels, their application is limited as they require native sGC to function. sGC activators on the other hand can function with heme-free sGC. Therefore, we tested the renoprotective effects of the sGC activator BAY 60-2770 that also functions with oxidized sGC in an acute kidney injury (AKI) model with transition to chronic kidney disease (CKD). In order to study the effects of BAY 60-2770 on gene expression in tubular cells, we used the proximal tubular cell line HK-2 as a model. To mimic the ischemia and fibrosis observed in vivo, we subjected HK-2 cells to hypoxia and TGF-β1 treatment, respectively. RNASeq analysis showed that BAY 60-2770 had a significant influence on the expression of genes involved in fibrosis, inflammation and tissue damage after both hypoxia and TGF-β1 treatments.

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Project description:Tubular epithelial cells (TECs) play a major role in potentiating renal fibrosis. While TGF-β1 is a key inducer of the pro-fibrotic phenotype in TECs, inhibition of TGF-β1 also blocks its protective effects, making this an unsuccessful strategy for treating renal fibrosis. Molecules induced by TGF-β1 to regulate TEC phenotype would serve as more specific and effective targets. To uncover such molecules, we performed high throughput analyses on TGF-β1 treated renal tubular epithelial cells (HKC). RNA sequencing identified 3565 genes and proteomics identified 74 proteins differentially regulated by TGF-β1 in HKC. We then selected 47 genes that were most upregulated in our in vitro datasets and that do not have well-characterized roles in renal fibrosis based on our review of the literature. We searched 8 publicly available datasets containing transcriptomic data from diseased human kidneys for gene expression patterns of the selected genes and found that MARCKS and DOCK2 were amongst the most consistently upregulated in the human datasets. In HKC, MARCKS knockdown decreased TGF-β1 mediated upregulation of pro-fibrotic markers, while DOCK2 knockdown had the opposite effect. Our data suggests novel roles for these proteins in renal TECs as potential promoters the pro-fibrotic phenotype.

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