title: Genome Therapy of Myotonic Dystrophy Type 1 iPS Cells for Development of Autologous Stem Cell Therapy.
abstract: Myotonic dystrophy type 1 (DM1) is caused by expanded Cytosine-Thymine-Guanine (CTG) repeats in the 3'-untranslated region (3' UTR) of the Dystrophia myotonica protein kinase (DMPK) gene, for which there is no effective therapy. The objective of this study is to develop genome therapy in human DM1 induced pluripotent stem (iPS) cells to eliminate mutant transcripts and reverse the phenotypes for developing autologous stem cell therapy. The general approach involves targeted insertion of polyA signals (PASs) upstream of DMPK CTG repeats, which will lead to premature termination of transcription and elimination of toxic mutant transcripts. Insertion of PASs was mediated by homologous recombination triggered by site-specific transcription activator-like effector nuclease (TALEN)-induced double-strand break. We found genome-treated DM1 iPS cells continue to maintain pluripotency. The insertion of PASs led to elimination of mutant transcripts and complete disappearance of nuclear RNA foci and reversal of aberrant splicing in linear-differentiated neural stem cells, cardiomyocytes, and teratoma tissues. In conclusion, genome therapy by insertion of PASs upstream of the expanded DMPK CTG repeats prevented the production of toxic mutant transcripts and reversal of phenotypes in DM1 iPS cells and their progeny. These genetically-treated iPS cells will have broad clinical application in developing autologous stem cell therapy for DM1.
authors.affiliation: Department of Neurology, University of Florida, College of Medicine, Gainesville, Florida, USA.|The Evelyn L & William F. McKnight Brain Institute, University of Florida, Florida, USA.
authors.forename: Yuanzheng
authors.initials: Y
authors.lastname: Gao
authors.affiliation: Department of Neurology, University of Florida, College of Medicine, Gainesville, Florida, USA.|The Evelyn L & William F. McKnight Brain Institute, University of Florida, Florida, USA.|Department of Neurology, the First Affiliated Hospital of Chongqing Medical University, Chongqing, China.
authors.forename: Xiuming
authors.initials: X
authors.lastname: Guo
authors.affiliation: Center for Cellular Reprogramming, University of Florida, College of Medicine, Gainesville, Florida, USA.|Department of Pathology, Immunology & Laboratory Medicine, University of Florida, College of Medicine, Gainesville, Florida, USA.
authors.forename: Katherine
authors.initials: K
authors.lastname: Santostefano
authors.affiliation: Department of Neurology, University of Florida, College of Medicine, Gainesville, Florida, USA.|The Evelyn L & William F. McKnight Brain Institute, University of Florida, Florida, USA.|Department of Neurology, the First Affiliated Hospital of Zhengzhou University, Henan, China.
authors.forename: Yanlin
authors.initials: Y
authors.lastname: Wang
authors.affiliation: Center for NeuroGenetics, University of Florida, College of Medicine, Gainesville, Florida, USA.|Molecular Genetics and Microbiology, College of Medicine, Gainesville, Florida, USA.
authors.forename: Tammy
authors.initials: T
authors.lastname: Reid
authors.affiliation: Department of Neurology, University of Florida, College of Medicine, Gainesville, Florida, USA.|The Evelyn L & William F. McKnight Brain Institute, University of Florida, Florida, USA.
authors.forename: Desmond
authors.initials: D
authors.lastname: Zeng
authors.affiliation: Center for Cellular Reprogramming, University of Florida, College of Medicine, Gainesville, Florida, USA.|Department of Pathology, Immunology & Laboratory Medicine, University of Florida, College of Medicine, Gainesville, Florida, USA.
authors.forename: Naohiro
authors.initials: N
authors.lastname: Terada
authors.affiliation: Department of Neurology, University of Florida, College of Medicine, Gainesville, Florida, USA.|The Evelyn L & William F. McKnight Brain Institute, University of Florida, Florida, USA.|Center for Cellular Reprogramming, University of Florida, College of Medicine, Gainesville, Florida, USA.|Center for NeuroGenetics, University of Florida, College of Medicine, Gainesville, Florida, USA.
authors.forename: Tetsuo
authors.initials: T
authors.lastname: Ashizawa
authors.affiliation: Department of Neurology, University of Florida, College of Medicine, Gainesville, Florida, USA.|The Evelyn L & William F. McKnight Brain Institute, University of Florida, Florida, USA.|Center for Cellular Reprogramming, University of Florida, College of Medicine, Gainesville, Florida, USA.|Center for NeuroGenetics, University of Florida, College of Medicine, Gainesville, Florida, USA.|Department of Neuroscience, University of Florida, Gainesville, Florida, USA.
authors.forename: Guangbin
authors.initials: G
authors.lastname: Xia
mesh_terms: D000818:Animals; D002454:Cell Differentiation; D002467:Cell Nucleus / Q000378:metabolism; D002471:Cell Transformation, Neoplastic / Q000235:genetics / Q000378:metabolism; D004195:Disease Models, Animal; D018390:Gene Targeting; D056426:Genetic Loci; D015316:Genetic Therapy; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism*; D051379:Mice; D032383:Myocytes, Cardiac / Q000166:cytology / Q000378:metabolism; D009223:Myotonic Dystrophy / Q000235:genetics* / Q000378:metabolism* / Q000628:therapy; D065636:Myotonin-Protein Kinase / Q000235:genetics; D011061:Poly A; D011485:Protein Binding; D012326:RNA Splicing; D033581:Stem Cell Transplantation; D013724:Teratoma / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D000069896:Transcription Activator-Like Effector Nucleases; D014182:Transplantation, Autologous; D018911:Trinucleotide Repeats
chemical_list: D011061:Poly A; D065636:Myotonin-Protein Kinase; D000069896:Transcription Activator-Like Effector Nucleases