Graded and pan-neural disease phenotypes of Rett Syndrome linked with dosage of functional MeCP2
Xiaoying Chen1(), Xu Han2, Bruno Blanchi3, Wuqiang Guan2, Weihong Ge3, Yong-Chun Yu2(), Yi E. Sun1()
1. Shanghai Institute of Stem Cell Research and Clinical Translation, Shanghai East Hospital, Tongji University School of Medicine, Shanghai 200120, China 2. State Key Laboratory of Medical Neurobiology and MOE Frontiers Center for Brain Science, Institutes of Brain Science, Jing’an District Centre Hospital of Shanghai, Fudan University, Shanghai 200032, China 3. Department of Psychiatry and Biobehavioral Sciences, David Geffen School of Medicine, UCLA, Los Angeles, CA 90095, USA
Rett syndrome (RTT) is a progressive neurodevelopmental disorder, mainly caused by mutations in MeCP2 and currently with no cure. We report here that neurons from R106W MeCP2 RTT human iPSCs as well as human embryonic stem cells after MeCP2 knockdown exhibit consistent and long-lasting impairment in maturation as indicated by impaired action potentials and passive membrane properties as well as reduced soma size and spine density. Moreover, RTT-inherent defects in neuronal maturation could be pan-neuronal and occurred in neurons with both dorsal and ventral forebrain features. Knockdown of MeCP2 led to more severe neuronal deficits as compared to RTT iPSC-derived neurons, which appeared to retain partial function. Strikingly, consistent deficits in nuclear size, dendritic complexity and circuitry-dependent spontaneous postsynaptic currents could only be observed in MeCP2 knockdown neurons but not RTT iPSC-derived neurons. Both neuron-intrinsic and circuitry-dependent deficits of MeCP2-deficient neurons could be fully or partially rescued by re-expression of wild type or T158M MeCP2, strengthening the dosage dependency of MeCP2 on disease phenotypes and also the partial function of the mutant. Our findings thus reveal stable neuronal maturation deficits and unexpectedly, graded sensitivities of neuron-inherent and neural transmission phenotypes towards the extent of MeCP2 deficiency, which is informative for future therapeutic development.
Corresponding Author(s):
Xiaoying Chen,Yong-Chun Yu,Yi E. Sun
引用本文:
. [J]. Protein & Cell, 2021, 12(8): 639-652.
Xiaoying Chen, Xu Han, Bruno Blanchi, Wuqiang Guan, Weihong Ge, Yong-Chun Yu, Yi E. Sun. Graded and pan-neural disease phenotypes of Rett Syndrome linked with dosage of functional MeCP2. Protein Cell, 2021, 12(8): 639-652.
RE Amir, IB Van den Veyver, M Wan, CQ Tran, U Francke, HY Zoghbi (1999) Rett syndrome is caused by mutations in X-linked MECP2, encoding methyl-CpG-binding protein 2. Nat Genet 23:185–188 https://doi.org/10.1038/13810
2
G Ananiev, EC Williams, H Li, Q Chang (2011) Isogenic pairs of wild type and mutant induced pluripotent stem cell (iPSC) lines from Rett syndrome patients as in vitro disease model. PLoS One 6: e25255 https://doi.org/10.1371/journal.pone.0025255
3
Y Asaka, DG Jugloff, L Zhang, JH Eubanks, RM Fitzsimonds (2006) Hippocampal synaptic plasticity is impaired in the Mecp2-null mouse model of Rett syndrome. Neurobiol Dis 21:217–227 https://doi.org/10.1016/j.nbd.2005.07.005
4
SJ Baudouin, J Gaudias, S Gerharz, L Hatstatt, K Zhou, P Punnakkal, KF Tanaka, W Spooren, R Hen, CI De Zeeuwet al. (2012) Shared synaptic pathophysiology in syndromic and nonsyndromic rodent models of autism. Science 338:128–132 https://doi.org/10.1126/science.1224159
5
Q Bu, A Wang, H Hamzah, A Waldman, K Jiang, Q Dong, R Li, J Kim, D Turner, Q Chang (2017) CREB signaling is involved in Rett syndrome pathogenesis. J Neurosci 37:3671–3685 https://doi.org/10.1523/JNEUROSCI.3735-16.2017
6
G Calfa, AK Percy, L Pozzo-Miller (2011) Experimental models of Rett syndrome based on Mecp2 dysfunction. Exp Biol Med (Maywood) 236:3–19 https://doi.org/10.1258/ebm.2010.010261
7
M Chahrour, SY Jung, C Shaw, X Zhou, ST Wong, J Qin, HY Zoghbi (2008) MeCP2, a key contributor to neurological disease, activates and represses transcription. Science 320:1224–1229 https://doi.org/10.1126/science.1153252
8
HT Chao, HY Zoghbi, C Rosenmund (2007) MeCP2 controls excitatory synaptic strength by regulating glutamatergic synapse number. Neuron 56:58–65 https://doi.org/10.1016/j.neuron.2007.08.018
9
CA Chapleau, GD Calfa, MC Lane, AJ Albertson, JL Larimore, S Kudo, DL Armstrong, AK Percy, L Pozzo-Miller (2009) Dendritic spine pathologies in hippocampal pyramidal neurons from Rett syndrome brain and after expression of Rett-associated MECP2 mutations. Neurobiol Dis 35:219–233 https://doi.org/10.1016/j.nbd.2009.05.001
10
X Chen, K Zhang, L Zhou, X Gao, J Wang, Y Yao, F He, Y Luo, Y Yu, S Liet al. (2016) Coupled electrophysiological recording and single cell transcriptome analyses revealed molecular mechanisms underlying neuronal maturation. Protein Cell 7:175–186 https://doi.org/10.1007/s13238-016-0247-8
11
Z Chen, X Ren, X Xu, X Zhang, Y Hui, Z Liu, L Shi, Y Fang, L Ma, Y Liuet al. (2018) Genetic engineering of human embryonic stem cells for precise cell fate tracing during human lineage development. Stem Cell Reports 11:1257–1271 https://doi.org/10.1016/j.stemcr.2018.09.014
12
X Chen, A Chanda, Y Ikeuchi, X Zhang, JV Goodman, NC Reddy, SP Majidi, DY Wu, SE Smith, A Godecet al. (2019) The transcriptional regulator SnoN promotes the proliferation of cerebellar granule neuron precursors in the postnatal mouse brain. J Neurosci 39:44–62 https://doi.org/10.1523/JNEUROSCI.0688-18.2018
13
AY Cheung, LM Horvath, D Grafodatskaya, P Pasceri, R Weksberg, A Hotta, L Carrel, J Ellis (2011) Isolation of MECP2-null Rett Syndrome patient hiPS cells and isogenic controls through X-chromosome inactivation. Hum Mol Genet 20:2103–2115 https://doi.org/10.1093/hmg/ddr093
14
VS Dani, SB Nelson (2009) Intact long-term potentiation but reduced connectivity between neocortical layer 5 pyramidal neurons in a mouse model of Rett syndrome. J Neurosci 29:11263–11270 https://doi.org/10.1523/JNEUROSCI.1019-09.2009
15
AD Ebert, J Yu, FF Jr Rose, VB Mattis, CL Lorson, JA Thomson, CN Svendsen (2009) Induced pluripotent stem cells from a spinal muscular atrophy patient. Nature 457:277–280 https://doi.org/10.1038/nature07677
16
N Farra, WB Zhang, P Pasceri, JH Eubanks, MW Salter, J Ellis (2012) Rett syndrome induced pluripotent stem cell-derived neurons reveal novel neurophysiological alterations. Mol Psychiatry 17:1261–1271 https://doi.org/10.1038/mp.2011.180
17
GM Jentarra, SL Olfers, SG Rice, N Srivastava, GE Homanics, M Blue, S Naidu, V Narayanan (2010) Abnormalities of cell packing density and dendritic complexity in the MeCP2 A140V mouse model of Rett syndrome/X-linked mental retardation. BMC Neurosci 11:19 https://doi.org/10.1186/1471-2202-11-19
18
XJ Li, X Zhang, MA Johnson, ZB Wang, T Lavaute, SC Zhang (2009) Coordination of sonic hedgehog and Wnt signaling determines ventral and dorsal telencephalic neuron types from human embryonic stem cells. Development 136:4055–4063 https://doi.org/10.1242/dev.036624
19
Y Li, H Wang, J Muffat, AW Cheng, DA Orlando, J Loven, SM Kwok, DA Feldman, HS Bateup, Q Gaoet al. (2013) Global transcriptional and translational repression in human-embryonic-stemcell-derived Rett syndrome neurons. Cell Stem Cell 13:446–458 https://doi.org/10.1016/j.stem.2013.09.001
20
Y Liu, JP Weick, H Liu, R Krencik, X Zhang, L Ma, GM Zhou, M Ayala, SC Zhang (2013) Medial ganglionic eminence-like cells derived from human embryonic stem cells correct learning and memory deficits. Nat Biotechnol 31:440–447 https://doi.org/10.1038/nbt.2565
21
S Luikenhuis, E Giacometti, CF Beard, R Jaenisch (2004) Expression of MeCP2 in postmitotic neurons rescues Rett syndrome in mice. Proc Natl Acad Sci USA 101:6033–6038 https://doi.org/10.1073/pnas.0401626101
22
MC Marchetto, C Carromeu, A Acab, D Yu, GW Yeo, Y Mu, G Chen, FH Gage, AR Muotri (2010) A model for neural development and treatment of Rett syndrome using human induced pluripotent stem cells. Cell 143:527–539 https://doi.org/10.1016/j.cell.2010.10.016
23
S Mekhoubad, C Bock, AS de Boer, E Kiskinis, A Meissner, K Eggan (2012) Erosion of dosage compensation impacts human iPSC disease modeling. Cell Stem Cell 10:595–609 https://doi.org/10.1016/j.stem.2012.02.014
24
M Mellen, P Ayata, S Dewell, S Kriaucionis, N Heintz (2012) MeCP2 binds to 5hmC enriched within active genes and accessible chromatin in the nervous system. Cell 151:1417–1430 https://doi.org/10.1016/j.cell.2012.11.022
25
P Moretti, JM Levenson, F Battaglia, R Atkinson, R Teague, B Antalffy, D Armstrong, O Arancio, JD Sweatt, HY Zoghbi (2006) Learning and memory and synaptic plasticity are impaired in a mouse model of Rett syndrome. J Neurosci 26:319–327 https://doi.org/10.1523/JNEUROSCI.2623-05.2006
26
X Nan, HH Ng, CA Johnson, CD Laherty, BM Turner, RN Eisenman, A Bird (1998) Transcriptional repression by the methyl-CpGbinding protein MeCP2 involves a histone deacetylase complex. Nature 393:386–389 https://doi.org/10.1038/30764
27
JL Neul, P Fang, J Barrish, J Lane, EB Caeg, EO Smith, H Zoghbi, A Percy, DG Glaze (2008) Specific mutations in methyl-CpGbinding protein 2 confer different severity in Rett syndrome. Neurology 70:1313–1321 https://doi.org/10.1212/01.wnl.0000291011.54508.aa
28
T Nikitina, X Shi, RP Ghosh, RA Horowitz-Scherer, JC Hansen, CL Woodcock (2007) Multiple modes of interaction between the methylated DNA binding protein MeCP2 and chromatin. Mol Cell Biol 27:864–877 https://doi.org/10.1128/MCB.01593-06
29
IH Park, N Arora, H Huo, N Maherali, T Ahfeldt, A Shimamura, MW Lensch, C Cowan, K Hochedlinger, GQ Daley (2008) Disease-specific induced pluripotent stem cells. Cell 134:877–886 https://doi.org/10.1016/j.cell.2008.07.041
30
K Plath, J Fang, SK Mlynarczyk-Evans, R Cao, KA Worringer, H Wang, CC de la Cruz, AP Otte, B Panning, Y Zhang (2003) Role of histone H3 lysine 27 methylation in X inactivation. Science 300:131–135 https://doi.org/10.1126/science.1084274
31
O Pomp, O Dreesen, DF Leong, O Meller-Pomp, TT Tan, F Zhou, A Colman (2011) Unexpected X chromosome skewing during culture and reprogramming of human somatic cells can be alleviated by exogenous telomerase. Cell Stem Cell 9:156–165 https://doi.org/10.1016/j.stem.2011.06.004
32
P Przanowski, U Wasko, Z Zheng, J Yu, R Sherman, LJ Zhu, MJ McConnell, J Tushir-Singh, MR Green, S Bhatnagar (2018) Pharmacological reactivation of inactive X-linked Mecp2 in cerebral cortical neurons of living mice. Proc Natl Acad Sci USA 115:7991–7996 https://doi.org/10.1073/pnas.1803792115
33
S Ricciardi, F Ungaro, M Hambrock, N Rademacher, G Stefanelli, D Brambilla, A Sessa, C Magagnotti, A Bachi, E Giardaet al. (2012) CDKL5 ensures excitatory synapse stability by reinforcing NGL-1-PSD95 interaction in the postsynaptic compartment and is impaired in patient iPSC-derived neurons. Nat Cell Biol 14:911–923 https://doi.org/10.1038/ncb2566
GM Stettner, P Huppke, C Brendel, DW Richter, J Gartner, M Dutschmann (2007) Breathing dysfunctions associated with impaired control of postinspiratory activity in Mecp2-/y knockout mice. J Physiol 579:863–876 https://doi.org/10.1113/jphysiol.2006.119966
36
K Takahashi, K Tanabe, M Ohnuki, M Narita, T Ichisaka, K Tomoda, S Yamanaka (2007) Induction of pluripotent stem cells from adult human fibroblasts by defined factors. Cell 131:861–872 https://doi.org/10.1016/j.cell.2007.11.019
J Tchieu, E Kuoy, MH Chin, H Trinh, M Patterson, SP Sherman, O Aimiuwu, A Lindgren, S Hakimian, JA Zacket al. (2010) Female human iPSCs retain an inactive X chromosome. Cell Stem Cell 7:329–342 https://doi.org/10.1016/j.stem.2010.06.024
39
JA Thomson, J Itskovitz-Eldor, SS Shapiro, MA Waknitz, JJ Swiergiel, VS Marshall, JM Jones (1998) Embryonic stem cell lines derived from human blastocysts. Science 282:1145–1147 https://doi.org/10.1126/science.282.5391.1145
40
K Tomoda, K Takahashi, K Leung, A Okada, M Narita, NA Yamada, KE Eilertson, P Tsang, S Baba, MP Whiteet al. (2012) Derivation conditions impact X-inactivation status in female human induced pluripotent stem cells. Cell Stem Cell 11:91–99 https://doi.org/10.1016/j.stem.2012.05.019
41
SC Wu, Y Zhang (2010) Active DNA demethylation: many roads lead to Rome. Nat Rev Mol Cell Biol 11:607–620 https://doi.org/10.1038/nrm2950
42
SC Zhang, M Wernig, ID Duncan, O Brustle, JA Thomson (2001) In vitro differentiation of transplantable neural precursors from human embryonic stem cells. Nat Biotechnol 19:1129–1133 https://doi.org/10.1038/nbt1201-1129
43
J Zou, ML Maeder, P Mali, SM Pruett-Miller, S Thibodeau-Beganny, BK Chou, G Chen, Z Ye, IH Park, GQ Daleyet al. (2009) Gene targeting of a disease-related gene in human induced pluripotent stem and embryonic stem cells. Cell Stem Cell 5:97–110 https://doi.org/10.1016/j.stem.2009.05.023
