title: Dystrophin-deficient cardiomyocytes derived from human urine: new biologic reagents for drug discovery.
abstract: The ability to extract somatic cells from a patient and reprogram them to pluripotency opens up new possibilities for personalized medicine. Induced pluripotent stem cells (iPSCs) have been employed to generate beating cardiomyocytes from a patient's skin or blood cells. Here, iPSC methods were used to generate cardiomyocytes starting from the urine of a patient with Duchenne muscular dystrophy (DMD). Urine was chosen as a starting material because it contains adult stem cells called urine-derived stem cells (USCs). USCs express the canonical reprogramming factors c-myc and klf4, and possess high telomerase activity. Pluripotency of urine-derived iPSC clones was confirmed by immunocytochemistry, RT-PCR and teratoma formation. Urine-derived iPSC clones generated from healthy volunteers and a DMD patient were differentiated into beating cardiomyocytes using a series of small molecules in monolayer culture. Results indicate that cardiomyocytes retain the DMD patient's dystrophin mutation. Physiological assays suggest that dystrophin-deficient cardiomyocytes possess phenotypic differences from normal cardiomyocytes. These results demonstrate the feasibility of generating cardiomyocytes from a urine sample and that urine-derived cardiomyocytes retain characteristic features that might be further exploited for mechanistic studies and drug discovery.
authors.affiliation: Department of Physiology and Pharmacology, School of Medicine, Wake Forest University Health Sciences, Winston-Salem, NC, USA; Department of Rehabilitation Medicine, University of Washington, Seattle, WA, USA.
authors.forename: Xuan
authors.initials: X
authors.lastname: Guan
authors.affiliation: Department of Rehabilitation Medicine, University of Washington, Seattle, WA, USA; Institute for Stem Cell and Regenerative Medicine, University of Washington, Seattle, WA, USA.
authors.forename: David L
authors.initials: DL
authors.lastname: Mack
authors.affiliation: Department of Physiology and Biophysics, University of Washington, Seattle, WA, USA.
authors.forename: Claudia M
authors.initials: CM
authors.lastname: Moreno
authors.affiliation: Division of Cardiovascular Medicine, Medical College of Wisconsin, Milwaukee, WI, USA.
authors.forename: Jennifer L
authors.initials: JL
authors.lastname: Strande
authors.affiliation: Institute for Stem Cell and Regenerative Medicine, University of Washington, Seattle, WA, USA; Department of Biochemistry, University of Washington, Seattle, WA, USA.
authors.forename: Julie
authors.initials: J
authors.lastname: Mathieu
authors.affiliation: Wake Forest Institute for Regenerative Medicine, Wake Forest University, Winston-Salem, NC, USA; Key Laboratory of Pathobiology, Ministry of Education, Jilin University, Changchun, China.
authors.forename: Yingai
authors.initials: Y
authors.lastname: Shi
authors.affiliation: Wake Forest Institute for Regenerative Medicine, Wake Forest University, Winston-Salem, NC, USA.
authors.forename: Chad D
authors.initials: CD
authors.lastname: Markert
authors.affiliation: Fred Hutchinson Cancer Research Center, Seattle, WA, USA.
authors.forename: Zejing
authors.initials: Z
authors.lastname: Wang
authors.affiliation: Wake Forest Institute for Regenerative Medicine, Wake Forest University, Winston-Salem, NC, USA.
authors.forename: Guihua
authors.initials: G
authors.lastname: Liu
authors.affiliation: Division of Pediatric Pathology, Department of Pathology and Laboratory Medicine, Medical College of Wisconsin, Milwaukee, WI, USA.
authors.forename: Michael W
authors.initials: MW
authors.lastname: Lawlor
authors.affiliation: Wake Forest Institute for Regenerative Medicine, Wake Forest University, Winston-Salem, NC, USA.
authors.forename: Emily C
authors.initials: EC
authors.lastname: Moorefield
authors.affiliation: Wake Forest Institute for Regenerative Medicine, Wake Forest University, Winston-Salem, NC, USA.
authors.forename: Tara N
authors.initials: TN
authors.lastname: Jones
authors.affiliation: Institute for Stem Cell and Regenerative Medicine, University of Washington, Seattle, WA, USA; Department of Pathology, University of Washington, Seattle, WA, USA.
authors.forename: James A
authors.initials: JA
authors.lastname: Fugate
authors.affiliation: Wake Forest Institute for Regenerative Medicine, Wake Forest University, Winston-Salem, NC, USA.
authors.forename: Mark E
authors.initials: ME
authors.lastname: Furth
authors.affiliation: Institute for Stem Cell and Regenerative Medicine, University of Washington, Seattle, WA, USA; Department of Pathology, University of Washington, Seattle, WA, USA; Center for Cardiovascular Biology, University of Washington, Seattle, WA, USA; Department of Bioengineering, University of Washington, Seattle, WA, USA; Department of Medicine/Cardiology, University of Washington, Seattle, WA, USA.
authors.forename: Charles E
authors.initials: CE
authors.lastname: Murry
authors.affiliation: Institute for Stem Cell and Regenerative Medicine, University of Washington, Seattle, WA, USA; Department of Biochemistry, University of Washington, Seattle, WA, USA.
authors.forename: Hannele
authors.initials: H
authors.lastname: Ruohola-Baker
authors.affiliation: Wake Forest Institute for Regenerative Medicine, Wake Forest University, Winston-Salem, NC, USA.
authors.forename: Yuanyuan
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: Department of Physiology and Biophysics, University of Washington, Seattle, WA, USA.
authors.forename: Luis F
authors.initials: LF
authors.lastname: Santana
authors.affiliation: Department of Rehabilitation Medicine, University of Washington, Seattle, WA, USA; Institute for Stem Cell and Regenerative Medicine, University of Washington, Seattle, WA, USA. Electronic address: mkc8@uw.edu.
authors.forename: Martin K
authors.initials: MK
authors.lastname: Childers
mesh_terms: D000328:Adult; D000818:Animals; D016022:Case-Control Studies; D002454:Cell Differentiation / Q000502:physiology; D002478:Cells, Cultured; D055808:Drug Discovery; D016189:Dystrophin / Q000172:deficiency*; D005260:Female; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000473:pathology*; D000090062:Kruppel-Like Factor 4; D051741:Kruppel-Like Transcription Factors / Q000096:biosynthesis / Q000652:urine; D008297:Male; D051379:Mice; D016688:Mice, Inbred NOD; D016513:Mice, SCID; D020388:Muscular Dystrophy, Duchenne / Q000473:pathology* / Q000652:urine*; D032383:Myocytes, Cardiac / Q000166:cytology / Q000378:metabolism* / Q000473:pathology*; D016271:Proto-Oncogene Proteins c-myc / Q000096:biosynthesis / Q000652:urine; D019098:Telomerase / Q000652:urine; D055815:Young Adult
chemical_list: D016189:Dystrophin; C000715467:KLF4 protein, human; C000715468:Klf4 protein, mouse; D000090062:Kruppel-Like Factor 4; D051741:Kruppel-Like Transcription Factors; C489427:MYC protein, human; D016271:Proto-Oncogene Proteins c-myc; D019098:Telomerase