Genetic deletion of Pax2 and Pax8 in adult mice
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ABSTRACT:
PROVIDER: PRJNA593517 | ENA |
REPOSITORIES: ENA
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Project description:Background: The mammalian kidneys maintain salt and water homeostasis for proper electrolyte balance and hydration. As the glomerular filtrate passes through the nephron and into the renal medulla, electrolytes, water, and urea are reabsorbed through the concerted actions of solute carrier channels and aquaporins located at various positions along the nephron and in the outer and inner medulla. Renal epithelial cells develop from Pax2 positive proliferating stem cells that suppress Pax2 expression once differentiated into mature proximal and distal tubules, but continue to express the related Pax8 protein. The collecting tubules and renal medulla are derived from a Pax2 positive ureteric bud epithelia that continue to express Pax2 and Pax8 in adult kidneys. Despite the necessity for Pax2 in renal development, functions for Pax2 or Pax8 in adult renal epithelia have not been established. Methods: In this report, we deleted either Pax2, Pax8, or both genes in adult mice and examined the phenotypes and changes in gene expression patterns. The mechanism of Pax8 mediated activation of potential target genes was described in inner medullary collecting duct cells. Results: Mice with induced deletions of both Pax2 and Pax8 exhibit severe polyuria that can be attributed to significant changes in the expression of solute carriers, such as the urea transporter UTA1, and aquaporins within the inner and outer medulla. Furthermore, Pax8 expression is induced by high salt in collecting duct cells and activates the UTA1 gene by recruiting a histone methyltransferase complex to the promoter. Conclusions: These data uncover novel functions for Pax proteins, in adult renal epithelia, that are essential for retaining water and concentrating urine.
2019-12-05 | GSE141455 | GEO
Project description:PAX2 and PAX8 are transcription factors linked to autosomal dominant polycystic kidney disease (ADPKD). Based on observations in 3D in vitro models showing that knockdown of either PAX2 or PAX8 reduced cyst-like growth in ADPKD-derived epithelial cells, transcriptomic analysis was performed to investigate their downstream gene regulatory roles. RNA sequencing was conducted in two human ADPKD-derived cell lines, WT9-7 and WT9-12, following siRNA-mediated knockdown. In WT9-7 cells, PAX2, PAX8, or combined PAX2/PAX8 knockdowns were performed, whereas in WT9-12 cells only PAX8 knockdown was carried out due to minimal PAX2 expression. Differential expression and gene ontology enrichment analyses were conducted to identify distinct and overlapping biological pathways regulated by PAX2 and PAX8 in cystic epithelial cells.
2026-05-20 | GSE305691 | GEO
Project description:Pax2 and Pax8 are homologous transcription factors required for kidney development and medullary urine concentration. However, their function in proximal tubule homeostasis and response to acute kidney injury is unknown. Mice with proximal tubules consisting of a mosaic of wild-type and Pax2/8 mutant proximal tubules cells were generated. Gene expression of mutant and wild-type proximal tubule cells was compared under homeostatic conditions using single-nucleus RNA sequencing.
2024-11-15 | GSE278289 | GEO
Project description:PAX2 and PAX8 are homologous DNA-binding proteins that orchestrate kidney development, regulate urine concentration in the kidney medulla, and contribute to sensitivity to and recovery from acute kidney injury. In some contexts, Pax proteins can recruit KMT2C/D histone methyl-transferase complexes that deposit histone H3K4 methylation marks to promote transcription or recruit the Polycomb Repressor Complex to silence gene expression. However, the spectrum of targets and functions of Pax proteins in kidney physiology remains poorly defined. This project aimed to identify genes regulated by PAX proteins in the proximal tubule. Proximal tubule epithelial cells were derived from mice with floxed Pax2 and Pax8 alleles and expressing a constitutive tamoxifen-response CreERT2 (mice described in PMID: 32381599). During propagation, Pax2 was spontaneously recombined, yielding a Pax2-null, Pax8-conditional cell line which was named PT-22. In this submission, the PT-22 line was transduced with a Tet-on expression system linked to a hemagglutinin (HA)-tagged Pax8a transgene and a mixed population of successfully transduced cells selected. PAX8 was depleted from these modified PT-22 cells with tamoxifen, then rescued by inducing re-expression of Pax8a transgene with doxycycline. Gene expression was measured to determine gene targets altered by PAX8 re-expression.
2026-07-15 | GSE317090 | GEO
Project description:PAX2 and PAX8 are homologous DNA-binding proteins that orchestrate kidney development, regulate urine concentration in the kidney medulla, and contribute to sensitivity to and recovery from acute kidney injury. In some contexts, Pax proteins can recruit KMT2C/D histone methyl-transferase complexes that deposit histone H3K4 methylation marks to promote transcription or recruit the Polycomb Repressor Complex to silence gene expression. However, the spectrum of targets and functions of Pax proteins in kidney physiology remains poorly defined. This project aimed to identify genes regulated by PAX proteins in the proximal tubule. Proximal tubule epithelial cells were derived from mice with floxed Pax2 and Pax8 alleles and expressing a constitutive tamoxifen-response CreERT2 (mice described in PMID: 32381599). During propagation, Pax2 was spontaneously recombined, yielding a Pax2-null, Pax8-conditional cell line which was named PT-22. In the experiments in this submission, PAX8 was depleted from PT-22 cells with tamoxifen and Pax8 chromatin occupancy and histone marks (H3K4me3 and H3K27me3) were measured after 6 d using chromatin immunoprecipitation followed by sequencing (ChIP-seq).
2026-07-15 | GSE317088 | GEO
Project description:PAX2 and PAX8 are homologous DNA-binding proteins that orchestrate kidney development, regulate urine concentration in the kidney medulla, and contribute to sensitivity to and recovery from acute kidney injury. In some contexts, Pax proteins can recruit KMT2C/D histone methyl-transferase complexes that deposit histone H3K4 methylation marks to promote transcription or recruit the Polycomb Repressor Complex to silence gene expression. However, the spectrum of targets and functions of Pax proteins in kidney physiology remains poorly defined. This project aimed to identify genes regulated by PAX proteins in the proximal tubule. Proximal tubule epithelial cells were derived from mice with floxed Pax2 and Pax8 alleles and expressing a constitutive tamoxifen-response CreERT2 (mice described in PMID: 32381599). During propagation, Pax2 was spontaneously recombined, yielding a Pax2-null, Pax8-conditional cell line which was named PT-22. In the experiments in this submission, PAX8 was depleted from PT-22 cells with tamoxifen and chromatin accessibility was measured after 0, 3, and 24 d using Assay for Transposase-Accessible Chromatin sequencing (ATAC-seq).
2026-07-15 | GSE317087 | GEO
Project description:PAX2 and PAX8 are homologous DNA-binding proteins that orchestrate kidney development, regulate urine concentration in the kidney medulla, and contribute to sensitivity to and recovery from acute kidney injury. In some contexts, Pax proteins can recruit KMT2C/D histone methyl-transferase complexes that deposit histone H3K4 methylation marks to promote transcription or recruit the Polycomb Repressor Complex to silence gene expression. However, the spectrum of targets and functions of Pax proteins in kidney physiology remains poorly defined. This project aimed to identify genes regulated by PAX proteins in the proximal tubule. Proximal tubule epithelial cells were derived from mice with floxed Pax2 and Pax8 alleles and expressing a constitutive tamoxifen-response CreERT2 (mice described in PMID: 32381599). During propagation, Pax2 was spontaneously recombined, yielding a Pax2-null, Pax8-conditional cell line which was named PT-22. In the experiments in this submission, PAX8 was depleted from PT-22 cells with tamoxifen and gene expression was measured over time to identify likely activating and repressive targets of PAX8.
2026-07-15 | GSE317089 | GEO
Project description:Single nucleus RNA sequencing of mosiac Pax2 and Pax8 mutant proximal tubules
| PRJNA1166220 | ENA
Project description:Activation of lineage-specific gene expression programs is mediated by the sequence-specific recruitment of transcriptional coactivators to chromatin. This occurs via interaction with lineage specific DNA binding transcription factors. The lineage factor PAX8 drives essential gene expression in ovarian cancer cells and is required for tumor proliferation. However, the molecular details surrounding co-factor recruitment and specific activation of transcription by PAX8 remain unknow. Here, we identify an important functional interaction between PAX8 and the Switch/Sucrose Non-Fermentable (SWI/SNF) chromatin remodeling complex. We show that PAX8 can recruit SWI/SNF complexes to DNA, where they function to open chromatin and facilitate expression of PAX8 target genes. Genetic deletion of PAX8 results in loss of SWI/SNF from PAX8 bound enhancers, loss of expression of associated target genes, and reduced proliferation. These results can be phenocopied by pharmacological inhibition of SWI/SNF ATPase activity. These data indicate that PAX8 mediates the expression of an essential ovarian cancer proliferative program in part by the recruitment of chromatin remodelers.
2024-08-04 | GSE272992 | GEO
Project description:Activation of lineage-specific gene expression programs is mediated by the sequence-specific recruitment of transcriptional coactivators to chromatin. This occurs via interaction with lineage specific DNA binding transcription factors. The lineage factor PAX8 drives essential gene expression in ovarian cancer cells and is required for tumor proliferation. However, the molecular details surrounding co-factor recruitment and specific activation of transcription by PAX8 remain unknow. Here, we identify an important functional interaction between PAX8 and the Switch/Sucrose Non-Fermentable (SWI/SNF) chromatin remodeling complex. We show that PAX8 can recruit SWI/SNF complexes to DNA, where they function to open chromatin and facilitate expression of PAX8 target genes. Genetic deletion of PAX8 results in loss of SWI/SNF from PAX8 bound enhancers, loss of expression of associated target genes, and reduced proliferation. These results can be phenocopied by pharmacological inhibition of SWI/SNF ATPase activity. These data indicate that PAX8 mediates the expression of an essential ovarian cancer proliferative program in part by the recruitment of chromatin remodelers.
2024-08-04 | GSE273024 | GEO