Cure of ADPKD by Selection for Spontaneous Genetic Repair Events in Pkd1-Mutated iPS Cells

• Published in: PloS one Vol. 7; no. 2; p. e32018
• Main Authors: Cheng, Li-Tao, Nagata, Shogo, Hirano, Kunio, Yamaguchi, Shinpei, Horie, Shigeo, Ainscough, Justin, Tada, Takashi
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 09.02.2012; Public Library of Science (PLoS)
• Subjects: Analysis; Animals; Biology; Cell division; Cell Transplantation; Chimera; Chromosomes; Clonal deletion; Cloning; Cysts; Disorders; DNA Repair; Epigenetic inheritance; epigenetics; Gene deletion; Gene therapy; Genes; Genetic aspects; Genetic disorders; genomics; Genotype & phenotype; Genotypes; Health aspects; Homologous recombination; Homology; Hybridization; Immunoglobulins; Induced Pluripotent Stem Cells; Inhibitory postsynaptic potentials; Kidney diseases; Kidney transplantation; Kidneys; Medicine; Mice; Morphology; Mutation; Patients; Pluripotency; Polycystic kidney; Polycystic Kidney, Autosomal Dominant - genetics; Polycystic Kidney, Autosomal Dominant - therapy; Polymerase chain reaction; Recombination; Recombination, Genetic; Repair; Restoration; Somatic cells; Stem cell transplantation; Stem cells; Tissue engineering; Transplantation; Treatment Outcome; TRPP Cation Channels - genetics; Tumorigenesis; Kyoto Japan; Japan
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0032018

Induced pluripotent stem cells (iPSCs) generated by epigenetic reprogramming of personal somatic cells have limited therapeutic capacity for patients suffering from genetic disorders. Here we demonstrate restoration of a genomic mutation heterozygous for Pkd1 (polycystic kidney disease 1) deletion (Pkd1(+/-) to Pkd1(+/R+)) by spontaneous mitotic recombination. Notably, recombination between homologous chromosomes occurred at a frequency of 1~2 per 10,000 iPSCs. Southern blot hybridization and genomic PCR analyses demonstrated that the genotype of the mutation-restored iPSCs was indistinguishable from that of the wild-type cells. Importantly, the frequency of cyst generation in kidneys of adult chimeric mice containing Pkd1(+/R+) iPSCs was significantly lower than that of adult chimeric mice with parental Pkd1(+/-) iPSCs, and indistinguishable from that of wild-type mice. This repair step could be directly incorporated into iPSC development programmes prior to cell transplantation, offering an invaluable step forward for patients carrying a wide range of genetic disorders.