Disease characterization using LQTS-specific induced pluripotent stem cells.

AIMS Long QT syndrome (LQTS) is an inheritable and life-threatening disease; however, it is often difficult to determine disease characteristics in sporadic cases with novel mutations, and more precise analysis is necessary for the successful development of evidence-based clinical therapies. This study thus sought to better characterize ion channel cardiac disorders using induced pluripotent stem cells (iPSCs). METHODS AND RESULTS We reprogrammed somatic cells from a patient with sporadic LQTS and from controls, and differentiated them into cardiomyocytes through embryoid body (EB) formation. Electrophysiological analysis of the LQTS-iPSC-derived EBs using a multi-electrode array (MEA) system revealed a markedly prolonged field potential duration (FPD). The IKr blocker E4031 significantly prolonged FPD in control- and LQTS-iPSC-derived EBs and induced frequent severe arrhythmia only in LQTS-iPSC-derived EBs. The IKs blocker chromanol 293B did not prolong FPD in the LQTS-iPSC-derived EBs, but significantly prolonged FPD in the control EBs, suggesting the involvement of IKs disturbance in the patient. Patch-clamp analysis and immunostaining confirmed a dominant-negative role for 1893delC in IKs channels due to a trafficking deficiency in iPSC-derived cardiomyocytes and human embryonic kidney (HEK) cells. CONCLUSIONS This study demonstrated that iPSCs could be useful to characterize LQTS disease as well as drug responses in the LQTS patient with a novel mutation. Such analyses may in turn lead to future progress in personalized medicine.

[1]  R. Weksberg,et al.  Isolation of MECP2-null Rett Syndrome patient hiPS cells and isogenic controls through X-chromosome inactivation , 2011, Human molecular genetics.

[2]  S. Yuasa,et al.  G-CSF promotes the proliferation of developing cardiomyocytes in vivo and in derivation from ESCs and iPSCs. , 2010, Cell stem cell.

[3]  Divya Rajamohan,et al.  Drug evaluation in cardiomyocytes derived from human induced pluripotent stem cells carrying a long QT syndrome type 2 mutation , 2011, European heart journal.

[4]  Lei Yang,et al.  Patient-specific induced pluripotent stem cell derived models of LEOPARD syndrome , 2010, Nature.

[5]  J. Stockman.  Genetic Testing in the Long QT Syndrome: Development and Validation of an Efficient Approach to Genotyping in Clinical Practice , 2007 .

[6]  E. Bettiol,et al.  Developmental Changes in Cardiomyocytes Differentiated from Human Embryonic Stem Cells: A Molecular and Electrophysiological Approach , 2007, Stem cells.

[7]  George Q. Daley,et al.  Disease-Specific Induced Pluripotent Stem Cells , 2008, Cell.

[8]  Shinsuke Yuasa,et al.  Generation of induced pluripotent stem cells from human terminally differentiated circulating T cells. , 2010, Cell stem cell.

[9]  I. Panfoli,et al.  Evidence for Ectopic Aerobic ATP Production on C6 Glioma Cell Plasma Membrane , 2011, Cellular and Molecular Neurobiology.

[10]  S. Priori,et al.  Diagnostic value of epinephrine test for genotyping LQT1, LQT2, and LQT3 forms of congenital long QT syndrome. , 2004, Heart rhythm.

[11]  D. Roden,et al.  Refining repolarization reserve. , 2011, Heart rhythm.

[12]  András Varró,et al.  Slow Delayed Rectifier Potassium Current (IKs) and the Repolarization Reserve , 2007, Annals of noninvasive electrocardiology : the official journal of the International Society for Holter and Noninvasive Electrocardiology, Inc.

[13]  T. Ichisaka,et al.  Induction of Pluripotent Stem Cells from Adult Human Fibroblasts by Defined Factors , 2007, Cell.

[14]  Dan M. Roden,et al.  ACC/AHA/ESC 2006 guidelines for management of patients with ventricular arrhythmias and the prevention of sudden cardiac death: A report of the American College of Cardiology/American Heart Association Task Force and the European Society of Cardiology Committee for Practice Guidelines , 2006 .

[15]  S. Priori,et al.  Genetic testing in the long QT syndrome: development and validation of an efficient approach to genotyping in clinical practice. , 2005, JAMA.

[16]  Ronald A. Li,et al.  Electrophysiological properties of human induced pluripotent stem cells. , 2010, American journal of physiology. Cell physiology.

[17]  Thomas D Rea,et al.  Community approaches to improve resuscitation after out-of-hospital sudden cardiac arrest. , 2010, Circulation.

[18]  C. Antzelevitch,et al.  Cellular Basis for Complex T Waves and Arrhythmic Activity Following Combined IKr and IKs Block , 2001, Journal of cardiovascular electrophysiology.

[19]  Jianming Xu,et al.  Extracellular K+ Is a Prerequisite for the Function and Plasma Membrane Stability of HERG Channels , 2010, Circulation research.

[20]  H. Huikuri,et al.  Sudden death due to cardiac arrhythmias. , 2001, The New England journal of medicine.

[21]  Hiroshi Morita,et al.  The QT syndromes: long and short , 2008, The Lancet.

[22]  篠原 隆司,et al.  Induction of pluripotent stem cell cells from germ cells , 2012 .

[23]  LimorZwi,et al.  Cardiomyocyte Differentiation of Human Induced Pluripotent Stem Cells , 2009 .

[24]  E. Marbán.  Cardiac channelopathies , 2020, Nature.

[25]  Karl-Ludwig Laugwitz,et al.  Patient-specific induced pluripotent stem-cell models for long-QT syndrome. , 2010, New England Journal of Medicine.

[26]  Susan Lindquist,et al.  Generation of Isogenic Pluripotent Stem Cells Differing Exclusively at Two Early Onset Parkinson Point Mutations , 2011, Cell.

[27]  Wojciech Zareba,et al.  Long QT Syndrome. , 1992, Current problems in cardiology.

[28]  Jonathan A. Bernstein,et al.  Using iPS cells to investigate cardiac phenotypes in patients with Timothy Syndrome , 2011, Nature.

[29]  T. Ichisaka,et al.  Induction of Pluripotent Stem Cells From Adult Human Fibroblasts by Defined Factors , 2008 .

[30]  K. Summers,et al.  Molecular genetics of long QT syndrome. , 2010, Molecular genetics and metabolism.

[31]  R. Stewart,et al.  Induced Pluripotent Stem Cell Lines Derived from Human Somatic Cells , 2007, Science.

[32]  M. Ackerman,et al.  Epinephrine QT Stress Testing in the Evaluation of Congenital Long-QT Syndrome: Diagnostic Accuracy of the Paradoxical QT Response , 2005, Circulation.

[33]  S. Priori,et al.  Low penetrance in the long-QT syndrome: clinical impact. , 1999, Circulation.

[34]  田中 智文,et al.  In vitro pharmacologic testing using human induced pluripotent stem cell-derived cardiomyocytes , 2012 .

[35]  Lior Gepstein,et al.  Modelling the long QT syndrome with induced pluripotent stem cells , 2011, Nature.

[36]  R. Virmani,et al.  Sudden cardiac death. , 1987, Human pathology.

[37]  World Medical Association Declaration of Helsinki. Recommendations guiding physicians in biomedical research involving human subjects. , 1997, Cardiovascular research.