Adenine base editor corrected ADPKD point mutations in hiPSCs and kidney organoids

Area of Science:

• Genetics
• Molecular Biology
• Regenerative Medicine

Background:

• Autosomal dominant polycystic kidney disease (ADPKD) is a genetic disorder caused by PKD1 mutations, leading to kidney failure.
• Current treatments for ADPKD are limited, and no gene therapy approaches exist to correct the underlying genetic defects.

Purpose of the Study:

• To investigate the potential of adenine base editor (ABE) technology for correcting specific PKD1 mutations associated with ADPKD.
• To develop and validate an in vitro model of ADPKD using patient-derived induced pluripotent stem cells (iPSCs) and kidney organoids.
• To evaluate ABE-mediated gene editing as a therapeutic strategy for ADPKD.

Main Methods:

• Identified two ADPKD-associated mutation sites (c.1198 C>T and c.8311 G>A) in the PKD1 gene.
• Tested ABE variant correction efficiencies in reporter cell lines and generated iPSCs from a patient's peripheral blood mononuclear cells (PBMCs).
• Differentiated iPSCs into kidney organoids, induced cystic phenotypes using forskolin, and applied a dual AAV split-ABEmax system for gene editing.

Main Results:

• ABE variants demonstrated varying correction efficiencies for the targeted PKD1 mutations.
• Patient-derived iPSC kidney organoids exhibited ADPKD-like cystic expansion upon cAMP stimulation, which was prevented in ABE-corrected organoids.
• The dual AAV split-ABEmax system achieved an average editing efficiency of approximately 6.56% in kidney organoids.

Conclusions:

• ABE single-base editing provides a viable framework for correcting pathogenic PKD1 mutations in ADPKD.
• ABE-corrected iPSC-derived kidney organoids successfully modeled ADPKD and demonstrated the therapeutic potential of gene editing.
• This study establishes a promising gene therapy strategy for ADPKD with potential for future clinical translation.