Please use this identifier to cite or link to this item: http://bura.brunel.ac.uk/handle/2438/30073
Title: De-Suppression of Mesenchymal Cell Identities and Variable Phenotypic Outcomes Associated with Knockout of Bbs1
Authors: Freke, GM
Martins, T
Davies, RJ
Beyer, T
Seda, M
Peskett, E
Haq, N
Prasai, A
Otto, G
Jeyabalan Srikaran, J
Hernandez, V
Diwan, GD
Russell, RB
Ueffing, M
Huranova, M
Boldt, K
Beales, PL
Jenkins, D
Keywords: Bardet–Biedl syndrome;primary cilia;epithelial-to-mesenchymal transition;kidney;collecting duct cells;Wnt signalling;fibrosis
Issue Date: 20-Nov-2023
Publisher: MDPI
Citation: Freke, G.M. e al. (2023) 'De-Suppression of Mesenchymal Cell Identities and Variable Phenotypic Outcomes Associated with Knockout of Bbs1', Cells, 12 (22), 2662, pp. 1 - 21. doi: 10.3390/cells12222662.
Abstract: Bardet–Biedl syndrome (BBS) is an archetypal ciliopathy caused by dysfunction of primary cilia. BBS affects multiple tissues, including the kidney, eye and hypothalamic satiety response. Understanding pan-tissue mechanisms of pathogenesis versus those which are tissue-specific, as well as gauging their associated inter-individual variation owing to genetic background and stochastic processes, is of paramount importance in syndromology. The BBSome is a membrane-trafficking and intraflagellar transport (IFT) adaptor protein complex formed by eight BBS proteins, including BBS1, which is the most commonly mutated gene in BBS. To investigate disease pathogenesis, we generated a series of clonal renal collecting duct IMCD3 cell lines carrying defined biallelic nonsense or frameshift mutations in Bbs1, as well as a panel of matching wild-type CRISPR control clones. Using a phenotypic screen and an unbiased multi-omics approach, we note significant clonal variability for all assays, emphasising the importance of analysing panels of genetically defined clones. Our results suggest that BBS1 is required for the suppression of mesenchymal cell identities as the IMCD3 cell passage number increases. This was associated with a failure to express epithelial cell markers and tight junction formation, which was variable amongst clones. Transcriptomic analysis of hypothalamic preparations from BBS mutant mice, as well as BBS patient fibroblasts, suggested that dysregulation of epithelial-to-mesenchymal transition (EMT) genes is a general predisposing feature of BBS across tissues. Collectively, this work suggests that the dynamic stability of the BBSome is essential for the suppression of mesenchymal cell identities as epithelial cells differentiate.
Description: Data Availability Statement: All novel datasets are available on request.
Supplementary Materials: The following supporting information can be downloaded at: https://www.mdpi.com/article/10.3390/cells12222662/s1 .
URI: https://bura.brunel.ac.uk/handle/2438/30073
DOI: https://doi.org/10.3390/cells12222662
Other Identifiers: ORCiD: Tina Beyer https://orcid.org/0000-0002-6302-8997
ORCiD: Marian Seda https://orcid.org/0000-0002-5634-3585
ORCiD: Naila Haq https://orcid.org/0000-0003-1887-5449
ORCiD: Georg Otto https://orcid.org/0000-0002-3929-948X
ORCiD: Jeshmi Jeyabalan Srikaran https://orcid.org/0000-0002-0195-8478
ORCiD: Victor Hernandez https://orcid.org/0000-0002-4424-1848
ORCiD: Marius Ueffing https://orcid.org/0000-0003-2209-2113
ORCiD: Martina Huranova https://orcid.org/0000-0002-4403-1146
ORCiD: Karsten Boldt https://orcid.org/0000-0002-2693-689X
ORCiD: Philip L. Beales https://orcid.org/0000-0002-9164-9782
2662
Appears in Collections:Dept of Life Sciences Research Papers

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