Modelling renal defects in Bardet-Biedl syndrome patients using human iPS cells
Bardet-Biedl syndrome (BBS) is a ciliopathy with pleiotropic effects on multiple tissues, including the kidney. Here we have compared renal differentiation of iPS cells from healthy and BBS donors. High content image analysis of WT1-expressing kidney progenitors showed that cell proliferation, diffe...
| Main Authors: | , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2023-06-01
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| Series: | Frontiers in Cell and Developmental Biology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fcell.2023.1163825/full |
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| author | James Williams Chloe Hurling Sabrina Munir Peter Harley Carolina Barcellos Machado Ana-Maria Cujba Mario Alvarez-Fallas Davide Danovi Davide Danovi Ivo Lieberam Ivo Lieberam Rocio Sancho Philip Beales Fiona M. Watt |
| author_facet | James Williams Chloe Hurling Sabrina Munir Peter Harley Carolina Barcellos Machado Ana-Maria Cujba Mario Alvarez-Fallas Davide Danovi Davide Danovi Ivo Lieberam Ivo Lieberam Rocio Sancho Philip Beales Fiona M. Watt |
| author_sort | James Williams |
| collection | DOAJ |
| description | Bardet-Biedl syndrome (BBS) is a ciliopathy with pleiotropic effects on multiple tissues, including the kidney. Here we have compared renal differentiation of iPS cells from healthy and BBS donors. High content image analysis of WT1-expressing kidney progenitors showed that cell proliferation, differentiation and cell shape were similar in healthy, BBS1, BBS2, and BBS10 mutant lines. We then examined three patient lines with BBS10 mutations in a 3D kidney organoid system. The line with the most deleterious mutation, with low BBS10 expression, expressed kidney marker genes but failed to generate 3D organoids. The other two patient lines expressed near normal levels of BBS10 mRNA and generated multiple kidney lineages within organoids when examined at day 20 of organoid differentiation. However, on prolonged culture (day 27) the proximal tubule compartment degenerated. Introducing wild type BBS10 into the most severely affected patient line restored organoid formation, whereas CRISPR-mediated generation of a truncating BBS10 mutation in a healthy line resulted in failure to generate organoids. Our findings provide a basis for further mechanistic studies of the role of BBS10 in the kidney. |
| first_indexed | 2024-03-13T07:55:30Z |
| format | Article |
| id | doaj.art-d171a7913adc46668842c8c82971940f |
| institution | Directory Open Access Journal |
| issn | 2296-634X |
| language | English |
| last_indexed | 2024-03-13T07:55:30Z |
| publishDate | 2023-06-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Cell and Developmental Biology |
| spelling | doaj.art-d171a7913adc46668842c8c82971940f2023-06-02T06:01:05ZengFrontiers Media S.A.Frontiers in Cell and Developmental Biology2296-634X2023-06-011110.3389/fcell.2023.11638251163825Modelling renal defects in Bardet-Biedl syndrome patients using human iPS cellsJames Williams0Chloe Hurling1Sabrina Munir2Peter Harley3Carolina Barcellos Machado4Ana-Maria Cujba5Mario Alvarez-Fallas6Davide Danovi7Davide Danovi8Ivo Lieberam9Ivo Lieberam10Rocio Sancho11Philip Beales12Fiona M. Watt13Centre for Gene Therapy and Regenerative Medicine, King’s College London, Guy’s Hospital, London, United KingdomCentre for Gene Therapy and Regenerative Medicine, King’s College London, Guy’s Hospital, London, United KingdomCentre for Gene Therapy and Regenerative Medicine, King’s College London, Guy’s Hospital, London, United KingdomCentre for Gene Therapy and Regenerative Medicine, King’s College London, Guy’s Hospital, London, United KingdomCentre for Gene Therapy and Regenerative Medicine, King’s College London, Guy’s Hospital, London, United KingdomCentre for Gene Therapy and Regenerative Medicine, King’s College London, Guy’s Hospital, London, United KingdomCentre for Gene Therapy and Regenerative Medicine, King’s College London, Guy’s Hospital, London, United KingdomCentre for Gene Therapy and Regenerative Medicine, King’s College London, Guy’s Hospital, London, United KingdomBit.bio, Babraham Research Campus, Cambridge, United KingdomCentre for Gene Therapy and Regenerative Medicine, King’s College London, Guy’s Hospital, London, United KingdomCentre for Developmental Neurobiology and MRC Centre for Neurodevelopmental Disorders, King’s College London, London, United KingdomCentre for Gene Therapy and Regenerative Medicine, King’s College London, Guy’s Hospital, London, United KingdomInstitute of Child Health, Genetic and Genomic Medicine, University College London, London, United KingdomCentre for Gene Therapy and Regenerative Medicine, King’s College London, Guy’s Hospital, London, United KingdomBardet-Biedl syndrome (BBS) is a ciliopathy with pleiotropic effects on multiple tissues, including the kidney. Here we have compared renal differentiation of iPS cells from healthy and BBS donors. High content image analysis of WT1-expressing kidney progenitors showed that cell proliferation, differentiation and cell shape were similar in healthy, BBS1, BBS2, and BBS10 mutant lines. We then examined three patient lines with BBS10 mutations in a 3D kidney organoid system. The line with the most deleterious mutation, with low BBS10 expression, expressed kidney marker genes but failed to generate 3D organoids. The other two patient lines expressed near normal levels of BBS10 mRNA and generated multiple kidney lineages within organoids when examined at day 20 of organoid differentiation. However, on prolonged culture (day 27) the proximal tubule compartment degenerated. Introducing wild type BBS10 into the most severely affected patient line restored organoid formation, whereas CRISPR-mediated generation of a truncating BBS10 mutation in a healthy line resulted in failure to generate organoids. Our findings provide a basis for further mechanistic studies of the role of BBS10 in the kidney.https://www.frontiersin.org/articles/10.3389/fcell.2023.1163825/fulliPSC (induced pluripotent stem cell)kidneyciliopathyCRISPRBardet-Biedl syndrome |
| spellingShingle | James Williams Chloe Hurling Sabrina Munir Peter Harley Carolina Barcellos Machado Ana-Maria Cujba Mario Alvarez-Fallas Davide Danovi Davide Danovi Ivo Lieberam Ivo Lieberam Rocio Sancho Philip Beales Fiona M. Watt Modelling renal defects in Bardet-Biedl syndrome patients using human iPS cells Frontiers in Cell and Developmental Biology iPSC (induced pluripotent stem cell) kidney ciliopathy CRISPR Bardet-Biedl syndrome |
| title | Modelling renal defects in Bardet-Biedl syndrome patients using human iPS cells |
| title_full | Modelling renal defects in Bardet-Biedl syndrome patients using human iPS cells |
| title_fullStr | Modelling renal defects in Bardet-Biedl syndrome patients using human iPS cells |
| title_full_unstemmed | Modelling renal defects in Bardet-Biedl syndrome patients using human iPS cells |
| title_short | Modelling renal defects in Bardet-Biedl syndrome patients using human iPS cells |
| title_sort | modelling renal defects in bardet biedl syndrome patients using human ips cells |
| topic | iPSC (induced pluripotent stem cell) kidney ciliopathy CRISPR Bardet-Biedl syndrome |
| url | https://www.frontiersin.org/articles/10.3389/fcell.2023.1163825/full |
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