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EZH2 specifically regulates ISL1 during embryonic urinary tract formation.

Enrico Mingardo^1,2, Jeshurun C Kalanithy^1,2, Gabriel Dworschak^2,3,4

¹Institute of Anatomy and Cell Biology, Medical Faculty, University of Bonn, 53115, Bonn, Germany.

Scientific Reports

|October 2, 2024

Summary

The gene ISL1 is crucial for urinary tract development. Its dysregulation, influenced by EZH2, is linked to classic bladder exstrophy (CBE), a condition causing bladder and urethral malformations.

Keywords:

Classic bladder exstrophy EZH2 Gene regulation HEK293 ISL1 Promoter Zebrafish

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Area of Science:

Developmental Biology
Genetics
Urology

Background:

The gene ISL1 is a known embryonic master control gene involved in genital development.
Genetic studies identified ISL1 as a key susceptibility gene for classic bladder exstrophy (CBE).
Previous research linked ISL1 deletion in mice to epispadias-like phenotypes.

Purpose of the Study:

To investigate the regulatory mechanisms of ISL1 in urinary tract formation.
To identify the role of EZH2 in ISL1 regulation and its association with CBE.
To explore the genetic basis of classic bladder exstrophy.

Main Methods:

Genome-wide association studies (GWAS) meta-analysis to identify significant markers near ISL1.
In silico analysis to predict regulatory effects of genetic markers.
Identification and characterization of a novel intragenic promoter for ISL1.
EZH2 silencing experiments in HEK cells and CRISPR/Cas9 knockout in zebrafish.

Main Results:

A novel ISL1 promoter region containing an EZH2 binding site was identified.
EZH2 silencing led to reduced ISL1 expression in HEK cells.
EZH2 knockout zebrafish larvae showed decreased ISL1 expression in the pronephric region.
EZH2 deficient zebrafish exhibited malformed nephric ducts, suggesting a role in urinary tract development.

Conclusions:

EZH2 is a critical regulator of ISL1 during urinary tract formation.
Tissue-specific dysregulation of ISL1, potentially mediated by EZH2, is implicated in the pathogenesis of classic bladder exstrophy.
This study provides novel insights into the molecular mechanisms underlying CBE.