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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; 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 (2019). 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. Kidney International, 95(2):375-387.

Abstract

Erythropoietin (Epo) is essential for erythropoiesis and is mainly produced by the fetal liver and the adult kidney following hypoxic stimulation. Epo regulation is commonly studied in hepatoma cell lines, but differences in Epo regulation between kidney and liver limit the understanding of Epo dysregulation in polycythaemia and anaemia. To overcome this limitation, we have generated a novel transgenic mouse model expressing Cre recombinase specifically in the active fraction of renal Epo-producing (REP) cells. Crossing with reporter mice confirmed the inducible and highly specific tagging of REP cells, located in the corticomedullary border region where there is a steep drop in oxygen bioavailability. A novel method was developed to selectively grow primary REP cells in culture and to generate immortalized clonal cell lines, called fibroblastoid atypical interstitial kidney (FAIK) cells. FAIK cells show very early hypoxia-inducible factor (HIF)-2α induction, which precedes Epo transcription. Epo induction in FAIK cells reverses rapidly despite ongoing hypoxia, suggesting a cell autonomous feedback mechanism. In contrast, HIF stabilizing drugs resulted in chronic Epo induction in FAIK cells. RNA sequencing of three FAIK cell lines derived from independent kidneys revealed a high degree of overlap and suggests that REP cells represent a unique cell type with properties of pericytes, fibroblasts, and neurons, known as telocytes. These novel cell lines may be helpful to investigate myofibroblast differentiation in chronic kidney disease and to elucidate the molecular mechanisms of HIF stabilizing drugs currently in phase III studies to treat anemia in end-stage kidney disease.

Abstract

Erythropoietin (Epo) is essential for erythropoiesis and is mainly produced by the fetal liver and the adult kidney following hypoxic stimulation. Epo regulation is commonly studied in hepatoma cell lines, but differences in Epo regulation between kidney and liver limit the understanding of Epo dysregulation in polycythaemia and anaemia. To overcome this limitation, we have generated a novel transgenic mouse model expressing Cre recombinase specifically in the active fraction of renal Epo-producing (REP) cells. Crossing with reporter mice confirmed the inducible and highly specific tagging of REP cells, located in the corticomedullary border region where there is a steep drop in oxygen bioavailability. A novel method was developed to selectively grow primary REP cells in culture and to generate immortalized clonal cell lines, called fibroblastoid atypical interstitial kidney (FAIK) cells. FAIK cells show very early hypoxia-inducible factor (HIF)-2α induction, which precedes Epo transcription. Epo induction in FAIK cells reverses rapidly despite ongoing hypoxia, suggesting a cell autonomous feedback mechanism. In contrast, HIF stabilizing drugs resulted in chronic Epo induction in FAIK cells. RNA sequencing of three FAIK cell lines derived from independent kidneys revealed a high degree of overlap and suggests that REP cells represent a unique cell type with properties of pericytes, fibroblasts, and neurons, known as telocytes. These novel cell lines may be helpful to investigate myofibroblast differentiation in chronic kidney disease and to elucidate the molecular mechanisms of HIF stabilizing drugs currently in phase III studies to treat anemia in end-stage kidney disease.

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Contributors:T. Buch and E. Campeau for the gift of plasmids, L.G. Cantley for the Terminator mouse strain, the Zurich Integrative Rodent Physiology facility, the Transgenic and Reproductive Techniques group of the University of Lausanne, the Functional Genomics Center Zurich, and the Center for Microscopy and Image Analysis for expert contributions to this wor
Item Type:Journal Article, refereed, original work
Communities & Collections:04 Faculty of Medicine > University Children's Hospital Zurich > Medical Clinic
04 Faculty of Medicine > Institute of Physiology
07 Faculty of Science > Institute of Physiology
Dewey Decimal Classification:570 Life sciences; biology
610 Medicine & health
Uncontrolled Keywords:Nephrology
Language:English
Date:1 February 2019
Deposited On:12 Feb 2019 16:53
Last Modified:12 Feb 2019 16:59
Publisher:Elsevier
ISSN:0085-2538
OA Status:Closed
Free access at:Publisher DOI. An embargo period may apply.
Publisher DOI:https://doi.org/10.1016/j.kint.2018.08.043
PubMed ID:30502050
Project Information:
  • : FunderSNSF
  • : Grant ID31003A_165679
  • : Project TitleHydroxylation-dependent functions of OTUB1 in oxygen physiology
  • : FunderSNSF
  • : Grant ID205321_153523
  • : Project TitleHR-Kidney - High Resolution 3D Functional Anatomy Database of the Kidney
  • : FunderNCCR Kidney.CH
  • : Grant ID
  • : Project Title
  • : Project Websitehttp://www.nccr-kidney.ch/
  • : FunderHartmann Müller-Stiftung
  • : Grant ID
  • : Project Title
  • : FunderEU’s seventh FP for research
  • : Grant IDgrant agreement no. 608847
  • : Project Title

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