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

• DOI: 10.1007/s44307-024-00026-8
• Publication Date: 2024-06-11T06:01:50Z
• Authors: Jingwen Wang; Yanling Qiu; Lei Zhang; Xinyao Zhou; Sihui Hu; Qianyi Liu; Sisi Yin; Zehong Su; Simiao Liu; Haiying Liu; Xueqing Wu; Junjiu Huang
• Keywords: Organoid; Base (topology)

Autosomal dominant polycystic kidney disease (ADPKD) is a genetic disorder caused primarily by mutations in the PKD1 gene, resulting formation of numerous cysts and eventually failure. However, there are currently no gene therapy studies aimed at correcting mutations. In this study, we identified two mutation sites associated with ADPKD, c.1198 (C > T) c.8311 (G A), which could potentially be corrected adenine base editor (ABE). The correction efficiencies different ABE variants were tested using HEK293T-PKD1 A) reporter cell lines. We then generated induced pluripotent stem cells (iPSCsmut/WT) from peripheral blood mononuclear (PBMCs) heterozygous patient to develop model. Since iPSCsmut/WT did not exhibit typical phenotype status, differentiation into organoids vitro led expression organ-specific marker proteins. Stimulation cAMP signaling forskolin resulted cystic expansion renal epithelial tissue iPSCmut/WT-derived organoids, resembling observed ADPKD patients. differentiated ABE-corrected iPSCs display phenotype. Furthermore, used dual AAV split-ABEmax system as therapeutic strategy achieved an average editing efficiency approximately 6.56% organoids. Overall, study provides framework for targeting through single-base editing, offering promising prospects future interventions.