Hnf1b renal expression directed by a distal enhancer responsive to Pax8
Abstract Xenopus provides a simple and efficient model system to study nephrogenesis and explore the mechanisms causing renal developmental defects in human. Hnf1b (hepatocyte nuclear factor 1 homeobox b), a gene whose mutations are the most commonly identified genetic cause of developmental kidney...
| Main Authors: | , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Nature Portfolio
2022-11-01
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| Series: | Scientific Reports |
| Online Access: | https://doi.org/10.1038/s41598-022-21171-x |
| _version_ | 1797983916053757952 |
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| author | L. Goea I. Buisson V. Bello A. Eschstruth M. Paces-Fessy R. Le Bouffant A. Chesneau S. Cereghini J. F. Riou M. Umbhauer |
| author_facet | L. Goea I. Buisson V. Bello A. Eschstruth M. Paces-Fessy R. Le Bouffant A. Chesneau S. Cereghini J. F. Riou M. Umbhauer |
| author_sort | L. Goea |
| collection | DOAJ |
| description | Abstract Xenopus provides a simple and efficient model system to study nephrogenesis and explore the mechanisms causing renal developmental defects in human. Hnf1b (hepatocyte nuclear factor 1 homeobox b), a gene whose mutations are the most commonly identified genetic cause of developmental kidney disease, is required for the acquisition of a proximo-intermediate nephron segment in Xenopus as well as in mouse. Genetic networks involved in Hnf1b expression during kidney development remain poorly understood. We decided to explore the transcriptional regulation of Hnf1b in the developing Xenopus pronephros and mammalian renal cells. Using phylogenetic footprinting, we identified an evolutionary conserved sequence (CNS1) located several kilobases (kb) upstream the Hnf1b transcription start and harboring epigenomic marks characteristics of a distal enhancer in embryonic and adult renal cells in mammals. By means of functional expression assays in Xenopus and mammalian renal cell lines we showed that CNS1 displays enhancer activity in renal tissue. Using CRISPR/cas9 editing in Xenopus tropicalis, we demonstrated the in vivo functional relevance of CNS1 in driving hnf1b expression in the pronephros. We further showed the importance of Pax8-CNS1 interaction for CNS1 enhancer activity allowing us to conclude that Hnf1b is a direct target of Pax8. Our work identified for the first time a Hnf1b renal specific enhancer and may open important perspectives into the diagnosis for congenital kidney anomalies in human, as well as modeling HNF1B-related diseases. |
| first_indexed | 2024-04-11T06:53:43Z |
| format | Article |
| id | doaj.art-a402f8c05af54aa7a35edda11baaa1d3 |
| institution | Directory Open Access Journal |
| issn | 2045-2322 |
| language | English |
| last_indexed | 2024-04-11T06:53:43Z |
| publishDate | 2022-11-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Scientific Reports |
| spelling | doaj.art-a402f8c05af54aa7a35edda11baaa1d32022-12-22T04:39:05ZengNature PortfolioScientific Reports2045-23222022-11-0112111710.1038/s41598-022-21171-xHnf1b renal expression directed by a distal enhancer responsive to Pax8L. Goea0I. Buisson1V. Bello2A. Eschstruth3M. Paces-Fessy4R. Le Bouffant5A. Chesneau6S. Cereghini7J. F. Riou8M. Umbhauer9Laboratoire de Biologie du Développement, CNRS, Institut de Biologie Paris Seine, IBPS, UMR7622, Sorbonne UniversitéLaboratoire de Biologie du Développement, CNRS, Institut de Biologie Paris Seine, IBPS, UMR7622, Sorbonne UniversitéLaboratoire de Biologie du Développement, CNRS, Institut de Biologie Paris Seine, IBPS, UMR7622, Sorbonne UniversitéLaboratoire de Biologie du Développement, CNRS, Institut de Biologie Paris Seine, IBPS, UMR7622, Sorbonne UniversitéLaboratoire de Biologie du Développement, CNRS, Institut de Biologie Paris Seine, IBPS, UMR7622, Sorbonne UniversitéLaboratoire de Biologie du Développement, CNRS, Institut de Biologie Paris Seine, IBPS, UMR7622, Sorbonne UniversitéParis-Saclay Institute of Neuroscience, CNRS, CERTO-Retina France, Université Paris-SaclayLaboratoire de Biologie du Développement, CNRS, Institut de Biologie Paris Seine, IBPS, UMR7622, Sorbonne UniversitéLaboratoire de Biologie du Développement, CNRS, Institut de Biologie Paris Seine, IBPS, UMR7622, Sorbonne UniversitéLaboratoire de Biologie du Développement, CNRS, Institut de Biologie Paris Seine, IBPS, UMR7622, Sorbonne UniversitéAbstract Xenopus provides a simple and efficient model system to study nephrogenesis and explore the mechanisms causing renal developmental defects in human. Hnf1b (hepatocyte nuclear factor 1 homeobox b), a gene whose mutations are the most commonly identified genetic cause of developmental kidney disease, is required for the acquisition of a proximo-intermediate nephron segment in Xenopus as well as in mouse. Genetic networks involved in Hnf1b expression during kidney development remain poorly understood. We decided to explore the transcriptional regulation of Hnf1b in the developing Xenopus pronephros and mammalian renal cells. Using phylogenetic footprinting, we identified an evolutionary conserved sequence (CNS1) located several kilobases (kb) upstream the Hnf1b transcription start and harboring epigenomic marks characteristics of a distal enhancer in embryonic and adult renal cells in mammals. By means of functional expression assays in Xenopus and mammalian renal cell lines we showed that CNS1 displays enhancer activity in renal tissue. Using CRISPR/cas9 editing in Xenopus tropicalis, we demonstrated the in vivo functional relevance of CNS1 in driving hnf1b expression in the pronephros. We further showed the importance of Pax8-CNS1 interaction for CNS1 enhancer activity allowing us to conclude that Hnf1b is a direct target of Pax8. Our work identified for the first time a Hnf1b renal specific enhancer and may open important perspectives into the diagnosis for congenital kidney anomalies in human, as well as modeling HNF1B-related diseases.https://doi.org/10.1038/s41598-022-21171-x |
| spellingShingle | L. Goea I. Buisson V. Bello A. Eschstruth M. Paces-Fessy R. Le Bouffant A. Chesneau S. Cereghini J. F. Riou M. Umbhauer Hnf1b renal expression directed by a distal enhancer responsive to Pax8 Scientific Reports |
| title | Hnf1b renal expression directed by a distal enhancer responsive to Pax8 |
| title_full | Hnf1b renal expression directed by a distal enhancer responsive to Pax8 |
| title_fullStr | Hnf1b renal expression directed by a distal enhancer responsive to Pax8 |
| title_full_unstemmed | Hnf1b renal expression directed by a distal enhancer responsive to Pax8 |
| title_short | Hnf1b renal expression directed by a distal enhancer responsive to Pax8 |
| title_sort | hnf1b renal expression directed by a distal enhancer responsive to pax8 |
| url | https://doi.org/10.1038/s41598-022-21171-x |
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