title: Isogenic human pluripotent stem cell pairs reveal the role of a KCNH2 mutation in long-QT syndrome.
abstract: Patient-specific induced pluripotent stem cells (iPSCs) will assist research on genetic cardiac maladies if the disease phenotype is recapitulated in vitro. However, genetic background variations may confound disease traits, especially for disorders with incomplete penetrance, such as long-QT syndromes (LQTS). To study the LQT2-associated c.A2987T (N996I) KCNH2 mutation under genetically defined conditions, we derived iPSCs from a patient carrying this mutation and corrected it. Furthermore, we introduced the same point mutation in human embryonic stem cells (hESCs), generating two genetically distinct isogenic pairs of LQTS and control lines. Correction of the mutation normalized the current (IKr) conducted by the HERG channel and the action potential (AP) duration in iPSC-derived cardiomyocytes (CMs). Introduction of the same mutation reduced IKr and prolonged the AP duration in hESC-derived CMs. Further characterization of N996I-HERG pathogenesis revealed a trafficking defect. Our results demonstrated that the c.A2987T KCNH2 mutation is the primary cause of the LQTS phenotype. Precise genetic modification of pluripotent stem cells provided a physiologically and functionally relevant human cellular context to reveal the pathogenic mechanism underlying this specific disease phenotype.
authors.affiliation: Department of Anatomy and Embryology, Leiden University Medical Center, Leiden, The Netherlands.
authors.forename: Milena
authors.initials: M
authors.lastname: Bellin
authors.forename: Simona
authors.initials: S
authors.lastname: Casini
authors.forename: Richard P
authors.initials: RP
authors.lastname: Davis
authors.forename: Cristina
authors.initials: C
authors.lastname: D'Aniello
authors.forename: Jessica
authors.initials: J
authors.lastname: Haas
authors.forename: Dorien
authors.initials: D
authors.lastname: Ward-van Oostwaard
authors.forename: Leon G J
authors.initials: LG
authors.lastname: Tertoolen
authors.forename: Christian B
authors.initials: CB
authors.lastname: Jung
authors.forename: David A
authors.initials: DA
authors.lastname: Elliott
authors.forename: Andrea
authors.initials: A
authors.lastname: Welling
authors.forename: Karl-Ludwig
authors.initials: KL
authors.lastname: Laugwitz
authors.forename: Alessandra
authors.initials: A
authors.lastname: Moretti
authors.forename: Christine L
authors.initials: CL
authors.lastname: Mummery
mesh_terms: D000200:Action Potentials / Q000235:genetics; D000328:Adult; D002478:Cells, Cultured; D000072237:ERG1 Potassium Channel; D053595:Embryonic Stem Cells / Q000502:physiology; D051638:Ether-A-Go-Go Potassium Channels / Q000235:genetics* / Q000378:metabolism; D005260:Female; D000072576:Homeobox Protein Nkx-2.5; D018398:Homeodomain Proteins / Q000235:genetics; D006801:Humans; D057026:Induced Pluripotent Stem Cells; D008133:Long QT Syndrome / Q000235:genetics*; D009154:Mutation; D032383:Myocytes, Cardiac / Q000473:pathology / Q000502:physiology; D018408:Patch-Clamp Techniques; D010641:Phenotype; D039904:Pluripotent Stem Cells / Q000502:physiology; D021381:Protein Transport / Q000235:genetics; D014157:Transcription Factors / Q000235:genetics
chemical_list: D000072237:ERG1 Potassium Channel; D051638:Ether-A-Go-Go Potassium Channels; D000072576:Homeobox Protein Nkx-2.5; D018398:Homeodomain Proteins; C000606913:KCNH2 protein, human; C095552:NKX2-5 protein, human; D014157:Transcription Factors