Generation of renal Epo-producing cell lines by conditional gene tagging reveals rapid HIF-2 driven Epo kinetics, cell autonomous feedback regulation, and a telocyte phenotype

Imeri, Faik; Nolan, Karen A.; Bapst, Andreas M.; Santambrogio, Sara; Abreu-Rodríguez, Irene; Spielmann, Patrick; Pfundstein, Svende; Libertini, Silvana; Crowther, Lisa M.; Orlando, Ilaria M.C.; Dahl, Sophie L.; Keodara, Anna; Kuo, Willy; Kurtcuoglu, Vartan; Scholz, Carsten C.; Qi, Weihong; Hummler, Edith; Hoogewijs, David; Wenger, Roland H.

doi:10.1016/j.kint.2018.08.043

Cited by 41 publications

(90 citation statements)

References 74 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…In non-EPO-producing cells, increased DNA methylation has been observed at the EPO gene locus 186 , which might possibly occur in the REPCs in CKD 205 . In support of this theory, one study reported that 5-azacytidine increases EPO production by immortalized REPCs in culture 208 . However, methylation-based silencing of the EPO gene is inconsistent with the known modes of action of PHD inhibitors, which can rapidly reactivate EPO expression, and was not supported by direct analysis of the EPO promoter 208 .…”

Section: [H2] Extinction and Reactivation Of Erythropoietin Productiomentioning

confidence: 90%

“…In support of this theory, one study reported that 5-azacytidine increases EPO production by immortalized REPCs in culture 208 . However, methylation-based silencing of the EPO gene is inconsistent with the known modes of action of PHD inhibitors, which can rapidly reactivate EPO expression, and was not supported by direct analysis of the EPO promoter 208 . Given that EPO is a specific HIF-2 target gene, a switch from HIF-2α towards HIF-1α expression in REPCs could theoretically have the potential to decrease EPO production, as would altered PHD expression or a reduction in oxygen consumption by these specific cells.…”

Section: [H2] Extinction and Reactivation Of Erythropoietin Productiomentioning

confidence: 90%

“…These studies have indicated that REPCs are of neural crest origin and are contained within a lineage specified by expression of the transcription factor forkhead box protein D1 (FOXD1) 207 . In normal kidneys, most REPCs are positive for the cell surface markers platelet-derived growth factor receptor-β (PDGFR-β) and 5'-nucleotidase (CD73), lie closely adjacent to the peritubular capillaries, and manifest long cellular protrusions 202,207,208 . Interestingly, it has been proposed that REPCs or other cells derived from FOXD1-expressing precursors, rather than cells derived by transdifferentiation of the renal epithelium, are the source of the fibroblasts responsible for the fibrosis that characterizes progressive CKD 209 .…”

Section: [H2] Extinction and Reactivation Of Erythropoietin Productiomentioning

confidence: 99%

See 2 more Smart Citations

Mechanisms of hypoxia signalling: new implications for nephrology

2019

View full text Add to dashboard Cite

Hypoxia-inducible factors (HIFs) transduce transcriptional responses to hypoxia that involve hundreds to thousands of target genes. • The oxygen-sensitive signal regulating HIF activity is generated by 2-oxoglutarate-dependent dioxygenases that catalyze the hydroxylation of specific HIF prolyl and asparaginyl residues to inactivate HIF in the presence of oxygen. • Inhibition of the HIF prolyl hydroxylases by 2-oxoglutarate analogues mimics hypoxia and activates many, but not all, components of the HIF transcriptional response. • Erythropoietin production by cortical interstitial fibroblasts in the kidney is very sensitive to activation of the HIF pathway. • In diseased kidneys, erythropoietin production is reduced, but can be increased by HIF prolyl hydroxylase inhibitors. • Activation of HIF has the potential to generate many other renal and systemic effects that will require consideration when HIF prolyl hydroxylase inhibitors are used clinically.

show abstract

Section: [H2] Extinction and Reactivation Of Erythropoietin Productiomentioning

confidence: 90%

Section: [H2] Extinction and Reactivation Of Erythropoietin Productiomentioning

confidence: 90%

Section: [H2] Extinction and Reactivation Of Erythropoietin Productiomentioning

confidence: 99%

See 1 more Smart Citation

Mechanisms of hypoxia signalling: new implications for nephrology

2019

View full text Add to dashboard Cite

show abstract

“…In the murine cardiomyocytes, the knockdown of Hif3a has been associated with a very minor upregulation of EPO expression, but an upregulation of HIF-1α and HIF-2α mRNA was also observed [38]. No human kidney-derived cell lines with inducible EPO expression exist, while the derivation of a murine kidney cell line with inducible EPO expression was reported only recently [39].…”

Section: Discussionmentioning

confidence: 99%

A long hypoxia-inducible factor 3 isoform 2 is a transcription activator that regulates erythropoietin

Tolonen

Heikkilä

Malinen

et al. 2019

Cell. Mol. Life Sci.

View full text Add to dashboard Cite

Hypoxia-inducible factor (HIF), an αβ dimer, is the master regulator of oxygen homeostasis with hundreds of hypoxiainducible target genes. Three HIF isoforms differing in the oxygen-sensitive α subunit exist in vertebrates. While HIF-1 and HIF-2 are known transcription activators, HIF-3 has been considered a negative regulator of the hypoxia response pathway. However, the human HIF3A mRNA is subject to complex alternative splicing. It was recently shown that the long HIF-3α variants can form αβ dimers that possess transactivation capacity. Here, we show that overexpression of the long HIF-3α2 variant induces the expression of a subset of genes, including the erythropoietin (EPO) gene, while simultaneous downregulation of all HIF-3α variants by siRNA targeting a shared HIF3A region leads to downregulation of EPO and additional genes. EPO mRNA and protein levels correlated with HIF3A silencing and HIF-3α2 overexpression. Chromatin immunoprecipitation analyses showed that HIF-3α2 binding associated with canonical hypoxia response elements in the promoter regions of EPO. Luciferase reporter assays showed that the identified HIF-3α2 chromatin-binding regions were sufficient to promote transcription by all three HIF-α isoforms. Based on these data, HIF-3α2 is a transcription activator that directly regulates EPO expression.

show abstract

“…hepatocytes is necessary for Epo upregulation (Kapitsinou et al, 2010;Minamishima and Kaelin, 2010;Haase, 2013;Koury and Haase, 2015;Imeri et al, 2019), a critical feature to increasing red blood cell density. Of the small molecules tested here, AK9 is the most robust Epo inducer, at least in vivo.…”

Section: Figure 2 | Potency Of Ar Compounds In Hif Stabilization and mentioning

confidence: 99%

Rank Order of Small Molecule Induced Hypoxiamimesis to Prevent Retinopathy of Prematurity

Hoppe

Bolok

McCollum

et al. 2020

Front. Cell Dev. Biol.

View full text Add to dashboard Cite

Here we rank order small molecule inhibitors of hypoxia inducible factor (HIF) prolyl hydroxylases (PHDs) using severity of oxygen induced retinopathy (OIR) as an outcome measure. Dose response analyses in cell cultures of hepatoma (Hep3B), retinal Müller cells (MIO-M1) and primary retinal endothelial cells were conducted to evaluate potency by comparing dose to HIF-1,2 protein levels by western blotting. In vivo dose response was determined using the luciferase-transgene HIF reporter (luc-ODD). Each compound was placed in rank order by their ability to reduce neovascularization and capillary drop out in the OIR mouse model. An Epas1 KO confined to retinal Müller cells was used to determine whether successful protection by HIF stabilization requires HIF-2. Two candidate small molecules can prevent OIR by stabilizing HIF-1 to prevent oxygen induced growth attenuation and vascular obliteration. Müller cell HIF-2, the mediator of pathologic retinal angiogenesis, is not required for protection. The lack of dependence on Müller cell HIF-2 predicts that inhibition of HIF PHD will not drive pathological angiogenesis.

show abstract

Generation of renal Epo-producing cell lines by conditional gene tagging reveals rapid HIF-2 driven Epo kinetics, cell autonomous feedback regulation, and a telocyte phenotype

Cited by 41 publications

References 74 publications

Mechanisms of hypoxia signalling: new implications for nephrology

Mechanisms of hypoxia signalling: new implications for nephrology

A long hypoxia-inducible factor 3 isoform 2 is a transcription activator that regulates erythropoietin

Rank Order of Small Molecule Induced Hypoxiamimesis to Prevent Retinopathy of Prematurity

Contact Info

Product

Resources

About