Kidney Organoid Modeling of WT1 Mutations Reveals Key Regulatory Paths Underlying Podocyte Development

• Published in: Advanced science Vol. 11; no. 29; pp. e2308556 - n/a
• Main Authors: Wang, Gang, Wu, Hangdi, Zhai, Xiuwen, Zhang, Li, Zhang, Changming, Cheng, Chen, Xu, Xiaodong, Gao, Erzhi, Xiong, Xushen, Zhang, Jin, Liu, Zhihong
• Format: Journal Article
• Language: English
• Published: Germany John Wiley & Sons, Inc 01.08.2024; John Wiley and Sons Inc; Wiley
• Subjects: Binding sites; Cells; Gene expression; Genotype & phenotype; Humans; Induced Pluripotent Stem Cells - metabolism; Kidney - metabolism; kidney organoid; Kidneys; Mutation; Mutation - genetics; Organoids - metabolism; podocyte development; Podocytes - metabolism; scATAC‐seq; scRNA‐seq; WT1; WT1 Proteins - genetics; WT1 Proteins - metabolism; scRNA‐seq; scATAC‐seq; podocyte development; kidney organoid; WT1
• ISSN: 2198-3844; 2198-3844
• DOI: 10.1002/advs.202308556

Wilms tumor‐1(WT1) is a crucial transcription factor that regulates podocyte development. However, the epigenomic mechanism underlying the function of WT1 during podocyte development has yet to be fully elucidated. Here, single‐cell chromatin accessibility and gene expression maps of foetal kidneys and kidney organoids are generated. Functional implications of WT1‐targeted genes, which are crucial for the development of podocytes and the maintenance of their structure, including BMPER/PAX2/MAGI2 that regulates WNT signaling pathway, MYH9 that maintains actin filament organization and NPHS1 that modulates cell junction assembly are identified. To further illustrate the functional importance of WT1‐mediated transcriptional regulation during podocyte development, cultured and implanted patient‐derived kidney organoids derived from the Induced Pluripotent Stem Cell (iPSCs) of a patient with a heterozygous missense mutation in WT1 are generated. Results from single‐cell RNA sequencing (scRNA‐seq) and functional assays confirm that the WT1 mutation leads to delays in podocyte development and causes damage to cell structures, due to its failure to activate the targeting genes MAGI2, MYH9, and NPHS1. Notably, correcting the mutation in the patient iPSCs using CRISPR‐Cas9 gene editing rescues the podocyte phenotype. Collectively, this work elucidates the WT1‐related epigenomic landscape with respect to human podocyte development and identifies the disease‐causing role of a WT1 mutation. In the present study, the WT1‐related epigenomic landscape of human podocyte development is elucidated by single‐cell multiomic dissection of foetal kidney and kidney organoids, revealing a substantial epigenomic similarity between kidney organoids and their foetal kidney counterparts. Furthermore, cultured and implanted kidney organoids desirably model the podocyte phenotypes and reveal the disease‐causing role and the underlying mechanism of a newly identified WT1 mutation.