Neuronal maturation defect in induced pluripotent stem cells from patients with Rett syndrome

Rett syndrome (RTT) is one of the most prevalent female neurodevelopmental disorders that cause severe mental retardation. Mutations in methyl CpG binding protein 2 (MeCP2) are mainly responsible for RTT. Patients with classical RTT exhibit normal development until age 6–18 mo, at which point they become symptomatic and display loss of language and motor skills, purposeful hand movements, and normal head growth. Murine genetic models and postmortem human brains have been used to study the disease and enable the molecular dissection of RTT. In this work, we applied a recently developed reprogramming approach to generate a novel in vitro human RTT model. Induced pluripotent stem cells (iPSCs) were derived from RTT fibroblasts by overexpressing the reprogramming factors OCT4, SOX2, KLF4, and MYC. Intriguingly, whereas some iPSCs maintained X chromosome inactivation, in others the X chromosome was reactivated. Thus, iPSCs were isolated that retained a single active X chromosome expressing either mutant or WT MeCP2, as well as iPSCs with reactivated X chromosomes expressing both mutant and WT MeCP2. When these cells underwent neuronal differentiation, the mutant monoallelic or biallelelic RTT-iPSCs displayed a defect in neuronal maturation consistent with RTT phenotypes. Our in vitro model of RTT is an important tool allowing the further investigation of the pathophysiology of RTT and the development of the curative therapeutics.

[1]  Colin A. Johnson,et al.  Transcriptional repression by the methyl-CpG-binding protein MeCP2 involves a histone deacetylase complex , 1998, Nature.

[2]  H. Zoghbi,et al.  Rett syndrome is caused by mutations in X-linked MECP2, encoding methyl-CpG-binding protein 2 , 1999, Nature Genetics.

[3]  H. Zoghbi,et al.  Influence of mutation type and X chromosome inactivation on Rett syndrome phenotypes , 2000, Annals of neurology.

[4]  Jeffrey J. Clare,et al.  Distribution of voltage‐gated sodium channel α‐subunit and β‐subunit mRNAs in human hippocampal formation, cortex, and cerebellum , 2000 .

[5]  R. Jaenisch,et al.  Deficiency of methyl-CpG binding protein-2 in CNS neurons results in a Rett-like phenotype in mice , 2001, Nature Genetics.

[6]  Y. Makita,et al.  The role of different X-inactivation pattern on the variable clinical phenotype with Rett syndrome , 2001, Brain and Development.

[7]  A. Bird,et al.  A mouse Mecp2-null mutation causes neurological symptoms that mimic Rett syndrome , 2001, Nature Genetics.

[8]  Hengbin Wang,et al.  Role of Histone H3 Lysine 27 Methylation in X Inactivation , 2003, Science.

[9]  Andrew J. Grimm,et al.  RettBASE: The IRSA MECP2 variation database—a new mutation database in evolution , 2003, Human mutation.

[10]  A mutant form of MeCP2 protein associated with human Rett syndrome cannot be displaced from methylated DNA by notch in Xenopus embryos. , 2003, Molecular cell.

[11]  Juan I. Young,et al.  X-chromosome inactivation patterns are unbalanced and affect the phenotypic outcome in a mouse model of rett syndrome. , 2004, American journal of human genetics.

[12]  Rodney C. Samaco,et al.  Multiple pathways regulate MeCP2 expression in normal brain development and exhibit defects in autism-spectrum disorders. , 2004, Human molecular genetics.

[13]  Wei Cui,et al.  Differentiation of Human Embryonic Stem Cells to Neural Lineages in Adherent Culture by Blocking Bone Morphogenetic Protein Signaling , 2005, Stem cells.

[14]  U. Francke.  Mechanisms of Disease: neurogenetics of MeCP2 deficiency , 2006, Nature Clinical Practice Neurology.

[15]  Su-Chun Zhang,et al.  In vitro differentiation of neural precursors from human embryonic stem cells. , 2006, Methods in molecular biology.

[16]  M. Bieda,et al.  Integrated epigenomic analyses of neuronal MeCP2 reveal a role for long-range interaction with active genes , 2007, Proceedings of the National Academy of Sciences.

[17]  Huda Y. Zoghbi,et al.  The Story of Rett Syndrome: From Clinic to Neurobiology , 2007, Neuron.

[18]  N. Nag,et al.  Behavioral and anatomical abnormalities in Mecp2 mutant mice: A model for Rett syndrome , 2007, Neuroscience.

[19]  Chad A. Cowan,et al.  Marked differences in differentiation propensity among human embryonic stem cell lines , 2008, Nature Biotechnology.

[20]  George Q. Daley,et al.  Reprogramming of human somatic cells to pluripotency with defined factors , 2008, Nature.

[21]  K. Hochedlinger,et al.  Defining molecular cornerstones during fibroblast to iPS cell reprogramming in mouse. , 2008, Cell stem cell.

[22]  Stephen T. C. Wong,et al.  MeCP2, a Key Contributor to Neurological Disease, Activates and Represses Transcription , 2008, Science.

[23]  Hynek Wichterle,et al.  Induced Pluripotent Stem Cells Generated from Patients with ALS Can Be Differentiated into Motor Neurons , 2008, Science.

[24]  R. Rowntree,et al.  X-chromosome inactivation and epigenetic fluidity in human embryonic stem cells , 2008, Proceedings of the National Academy of Sciences.

[25]  Paul H Lerou,et al.  Generation of human-induced pluripotent stem cells , 2008, Nature Protocols.

[26]  G. Daley,et al.  Live cell imaging distinguishes bona fide human iPS cells from partially reprogrammed cells , 2009, Nature Biotechnology.

[27]  Jehyuk Lee,et al.  A Robust Approach to Identifying Tissue-Specific Gene Expression Regulatory Variants Using Personalized Human Induced Pluripotent Stem Cells , 2009, PLoS genetics.

[28]  A. Viale,et al.  Modeling Pathogenesis and Treatment of Familial Dysautonomia using Patient Specific iPSCs , 2009, Nature.

[29]  G. Mandel,et al.  Non–cell autonomous influence of MeCP2-deficient glia on neuronal dendritic morphology , 2009, Nature Neuroscience.

[30]  K. Hochedlinger,et al.  Epigenetic reprogramming and induced pluripotency , 2009, Development.

[31]  James A. Thomson,et al.  Induced pluripotent stem cells from a spinal muscular atrophy patient , 2009, Nature.

[32]  D. Harvey,et al.  Rett Syndrome Astrocytes Are Abnormal and Spread MeCP2 Deficiency through Gap Junctions , 2009, The Journal of Neuroscience.

[33]  Izumi Maezawa,et al.  Rett Syndrome Microglia Damage Dendrites and Synapses by the Elevated Release of Glutamate , 2010, The Journal of Neuroscience.

[34]  Jennifer A. Erwin,et al.  Derivation of Pre-X Inactivation Human Embryonic Stem Cells under Physiological Oxygen Concentrations , 2010, Cell.

[35]  Fred H. Gage,et al.  A Model for Neural Development and Treatment of Rett Syndrome Using Human Induced Pluripotent Stem Cells , 2010, Cell.

[36]  Tamar Dvash,et al.  Variations of X Chromosome Inactivation Occur in Early Passages of Female Human Embryonic Stem Cells , 2010, PloS one.

[37]  C. Lengner,et al.  Human embryonic stem cells with biological and epigenetic characteristics similar to those of mouse ESCs , 2010, Proceedings of the National Academy of Sciences.

[38]  K. Plath,et al.  Female human iPSCs retain an inactive X chromosome. , 2010, Cell stem cell.

[39]  H. Tsukamoto,et al.  MeCP2 controls an epigenetic pathway that promotes myofibroblast transdifferentiation and fibrosis. , 2010, Gastroenterology.

[40]  Krishanu Saha,et al.  Pluripotency and Cellular Reprogramming: Facts, Hypotheses, Unresolved Issues , 2010, Cell.

[41]  James A Thomson,et al.  Neural differentiation of human induced pluripotent stem cells follows developmental principles but with variable potency , 2010, Proceedings of the National Academy of Sciences.

[42]  N. Benvenisty,et al.  Meta-analysis of the heterogeneity of X chromosome inactivation in human pluripotent stem cells. , 2011, Stem cell research.

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