Altered Neurite Morphology and Cholinergic Function of Induced Pluripotent Stem Cell-derived Neurons from a Patient with Kleefstra Syndrome and Autism

Overview

Journal Transl Psychiatry

Specialties Psychiatry
Public Health

Date 2017 Jul 26

PMID 28742076

Citations 18

Authors

J Nagy

J Kobolak

S Berzsenyi

Z Abraham

H X Avci

I Bock

Z Bekes

B Hodoscsek

A Chandrasekaran

A Teglasi

P Dezso

B Kovanyi

E T Voros

L Fodor

T Szel

K Nemeth

A Balazs

A Dinnyes

B Lendvai

G Levay

V Roman

Affiliations

Soon will be listed here.

Abstract

The aim of the present study was to establish an in vitro Kleefstra syndrome (KS) disease model using the human induced pluripotent stem cell (hiPSC) technology. Previously, an autism spectrum disorder (ASD) patient with Kleefstra syndrome (KS-ASD) carrying a deleterious premature termination codon mutation in the EHMT1 gene was identified. Patient specific hiPSCs generated from peripheral blood mononuclear cells of the KS-ASD patient were differentiated into post-mitotic cortical neurons. Lower levels of EHMT1 mRNA as well as protein expression were confirmed in these cells. Morphological analysis on neuronal cells differentiated from the KS-ASD patient-derived hiPSC clones showed significantly shorter neurites and reduced arborization compared to cells generated from healthy controls. Moreover, density of dendritic protrusions of neuronal cells derived from KS-ASD hiPSCs was lower than that of control cells. Synaptic connections and spontaneous neuronal activity measured by live cell calcium imaging could be detected after 5 weeks of differentiation, when KS-ASD cells exhibited higher sensitivity of calcium responses to acetylcholine stimulation indicating a lower nicotinic cholinergic tone at baseline condition in KS-ASD cells. In addition, gene expression profiling of differentiated neuronal cells from the KS-ASD patient revealed higher expression of proliferation-related genes and lower mRNA levels of genes involved in neuronal maturation and migration. Our data demonstrate anomalous neuronal morphology, functional activity and gene expression in KS-ASD patient-specific hiPSC-derived neuronal cultures, which offers an in vitro system that contributes to a better understanding of KS and potentially other neurodevelopmental disorders including ASD.

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References

Kawai H, Zago W, Berg D . Nicotinic alpha 7 receptor clusters on hippocampal GABAergic neurons: regulation by synaptic activity and neurotrophins. J Neurosci. 2002; 22(18):7903-12. PMC: 6758091. View

Dravid S, Murray T . Spontaneous synchronized calcium oscillations in neocortical neurons in the presence of physiological [Mg(2+)]: involvement of AMPA/kainate and metabotropic glutamate receptors. Brain Res. 2004; 1006(1):8-17. DOI: 10.1016/j.brainres.2004.01.059. View

Douglas R, Martin K . Neuronal circuits of the neocortex. Annu Rev Neurosci. 2004; 27:419-51. DOI: 10.1146/annurev.neuro.27.070203.144152. View

Chung M, Dalton K, Alexander A, Davidson R . Less white matter concentration in autism: 2D voxel-based morphometry. Neuroimage. 2004; 23(1):242-51. DOI: 10.1016/j.neuroimage.2004.04.037. View

McAlonan G, Cheung V, Cheung C, Suckling J, Lam G, Tai K . Mapping the brain in autism. A voxel-based MRI study of volumetric differences and intercorrelations in autism. Brain. 2004; 128(Pt 2):268-76. DOI: 10.1093/brain/awh332. View

Zago W, Massey K, Berg D . Nicotinic activity stabilizes convergence of nicotinic and GABAergic synapses on filopodia of hippocampal interneurons. Mol Cell Neurosci. 2006; 31(3):549-59. DOI: 10.1016/j.mcn.2005.11.009. View

Vidal C, Nicolson R, DeVito T, Hayashi K, Geaga J, Drost D . Mapping corpus callosum deficits in autism: an index of aberrant cortical connectivity. Biol Psychiatry. 2006; 60(3):218-25. DOI: 10.1016/j.biopsych.2005.11.011. View

Kleefstra T, Brunner H, Amiel J, Oudakker A, Nillesen W, Magee A . Loss-of-function mutations in euchromatin histone methyl transferase 1 (EHMT1) cause the 9q34 subtelomeric deletion syndrome. Am J Hum Genet. 2006; 79(2):370-7. PMC: 1559478. DOI: 10.1086/505693. View

Liu Z, Neff R, Berg D . Sequential interplay of nicotinic and GABAergic signaling guides neuronal development. Science. 2006; 314(5805):1610-3. DOI: 10.1126/science.1134246. View

10.

Geschwind D, Levitt P . Autism spectrum disorders: developmental disconnection syndromes. Curr Opin Neurobiol. 2007; 17(1):103-11. DOI: 10.1016/j.conb.2007.01.009. View

11.

McKay B, Placzek A, Dani J . Regulation of synaptic transmission and plasticity by neuronal nicotinic acetylcholine receptors. Biochem Pharmacol. 2007; 74(8):1120-33. PMC: 2047292. DOI: 10.1016/j.bcp.2007.07.001. View

12.

Giocomo L, Hasselmo M . Neuromodulation by glutamate and acetylcholine can change circuit dynamics by regulating the relative influence of afferent input and excitatory feedback. Mol Neurobiol. 2007; 36(2):184-200. DOI: 10.1007/s12035-007-0032-z. View

13.

Schmittgen T, Livak K . Analyzing real-time PCR data by the comparative C(T) method. Nat Protoc. 2008; 3(6):1101-8. DOI: 10.1038/nprot.2008.73. View

14.

Chambers S, Fasano C, Papapetrou E, Tomishima M, Sadelain M, Studer L . Highly efficient neural conversion of human ES and iPS cells by dual inhibition of SMAD signaling. Nat Biotechnol. 2009; 27(3):275-80. PMC: 2756723. DOI: 10.1038/nbt.1529. View

15.

Kleefstra T, van Zelst-Stams W, Nillesen W, Cormier-Daire V, Houge G, Foulds N . Further clinical and molecular delineation of the 9q subtelomeric deletion syndrome supports a major contribution of EHMT1 haploinsufficiency to the core phenotype. J Med Genet. 2009; 46(9):598-606. DOI: 10.1136/jmg.2008.062950. View

16.

Heikkila T, Yla-Outinen L, Tanskanen J, Lappalainen R, Skottman H, Suuronen R . Human embryonic stem cell-derived neuronal cells form spontaneously active neuronal networks in vitro. Exp Neurol. 2009; 218(1):109-16. DOI: 10.1016/j.expneurol.2009.04.011. View

17.

Balemans M, Huibers M, Eikelenboom N, Kuipers A, van Summeren R, Pijpers M . Reduced exploration, increased anxiety, and altered social behavior: Autistic-like features of euchromatin histone methyltransferase 1 heterozygous knockout mice. Behav Brain Res. 2009; 208(1):47-55. DOI: 10.1016/j.bbr.2009.11.008. View

18.

Hutsler J, Zhang H . Increased dendritic spine densities on cortical projection neurons in autism spectrum disorders. Brain Res. 2009; 1309:83-94. DOI: 10.1016/j.brainres.2009.09.120. View

19.

Wegiel J, Kuchna I, Nowicki K, Imaki H, Wegiel J, Marchi E . The neuropathology of autism: defects of neurogenesis and neuronal migration, and dysplastic changes. Acta Neuropathol. 2010; 119(6):755-70. PMC: 2869041. DOI: 10.1007/s00401-010-0655-4. View

20.

Schumann C, Bloss C, Barnes C, Wideman G, Carper R, Akshoomoff N . Longitudinal magnetic resonance imaging study of cortical development through early childhood in autism. J Neurosci. 2010; 30(12):4419-27. PMC: 2859218. DOI: 10.1523/JNEUROSCI.5714-09.2010. View