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Streamlined identification of clinically and functionally relevant genetic regulators of lower tract urogenital development

ABSTRACT: Streamlined identification of clinically and functionally relevant genetic regulators of lower tract urogenital development

PROVIDER: PRJNA1013971 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR25944716.fastq.gz	Fastqsanger.gz
	SRR25944717.fastq.gz	Fastqsanger.gz
	SRR25944718.fastq.gz	Fastqsanger.gz
	SRR25944719.fastq.gz	Fastqsanger.gz
	SRR25944720.fastq.gz	Fastqsanger.gz

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Streamlined identification of clinically and functionally relevant genetic regulators of lower tract urogenital development

Project description:Congenital anomalies of the lower genitourinary (LGU) tract are frequently comorbid due to genetically linked developmental pathways. Genital phenotypes represent the second most common group of congenital structural anomalies in males. Genes involved in sexual differentiation are prime candidates for developmental anomalies of multiple LGU organs, but insufficient prospective screening tools have prevented the rapid identification of causative genes. Androgen signaling is among the most influential modulators of LGU development. The present study uses SpDamID technology in vivo to generate a comprehensive map of the pathways actively regulated by androgen receptor (AR) in the genitalia in the presence of the p300 co-activator, identifying WNT signaling as a highly enriched AR-regulated pathway in the genitalia. Transcription factor (TF) hits were then assayed for sexually dimorphic expression at two critical timepoints, and also cross-referenced to the DECIPHER database of clinically relevant copy number variations (CNVs) to identify 252 TFs exhibiting copy variation in patients with LGU phenotypes. A subset of 54 TFs was identified for which LGU phenotypes are statistically overrepresented as a proportion of total observed phenotypes. The 252 TF hitlist was then subjected to a functional screen to identify hits whose silencing affects genital mesenchymal growth rates. Overlap of these datasets results in a refined list of 133 transcription factors of both functional and clinical relevance to LGU development, 31 of which are top priority candidates, including the well-documented renal progenitor regulator, Sall1. Loss of Sall1 was examined in vivo and confirmed to be a powerful regulator of LGU development

2024-01-16 | GSE242628 | GEO

Streamlined identification of clinically and functionally relevant genetic regulators of lower-tract urogenital development.

Project description:Congenital anomalies of the lower genitourinary (LGU) tract are frequently comorbid due to genetically linked developmental pathways, and are among the most common yet most socially stigmatized congenital phenotypes. Genes involved in sexual differentiation are prime candidates for developmental anomalies of multiple LGU organs, but insufficient prospective screening tools have prevented the rapid identification of causative genes. Androgen signaling is among the most influential modulators of LGU development. The present study uses SpDamID technology in vivo to generate a comprehensive map of the pathways actively regulated by the androgen receptor (AR) in the genitalia in the presence of the p300 coactivator, identifying wingless/integrated (WNT) signaling as a highly enriched AR-regulated pathway in the genitalia. Transcription factor (TF) hits were then assayed for sexually dimorphic expression at two critical time points and also cross-referenced to a database of clinically relevant copy number variations to identify 252 TFs exhibiting copy variation in patients with LGU phenotypes. A subset of 54 TFs was identified for which LGU phenotypes are statistically overrepresented as a proportion of total observed phenotypes. The 252 TF hitlist was then subjected to a functional screen to identify hits whose silencing affects genital mesenchymal growth rates. Overlap of these datasets results in a refined list of 133 TFs of both functional and clinical relevance to LGU development, 31 of which are top priority candidates, including the well-documented renal progenitor regulator, Sall1. Loss of Sall1 was examined in vivo and confirmed to be a powerful regulator of LGU development.

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Project description:We will use micro-dissection and FACS techniques to isolate cell types from the developing lower urogenital region and perform a transcriptional analysis using RNA-SEQ. This analysis will determine the transcriptional profile of each cell type, identify compartment specific transcripts, compartment specific transcript isoforms and cell-type specific long-noncoding RNAs. In addition the unbiased nature of RNA-SEQ will potentially identify novel transcripts that have not been annotated in the database. Total RNA was obtained from FACS isolated embryonic day 17.5 bladders using Tg(Upk2-cre)6Xrw x Gt(ROSA)26Sortm4(ACTB-tdTomato,-EGFP)Luo transgenic mice. The long term goal is to generate a transcriptional atlas of developing lower urogenital region.

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