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P.135 Autism-associated mutations in SHANK2 increase synaptic connectivity and dendrite complexity in human neurons

Published online by Cambridge University Press:  27 June 2018

K Zaslavsky
Affiliation:
(Toronto)
W Zhang
Affiliation:
(Toronto)
E Deneault
Affiliation:
(Toronto)
M Zhao
Affiliation:
(Toronto)
DC Rodrigues
Affiliation:
(Toronto)
F McReady
Affiliation:
(Toronto)
PJ Ross
Affiliation:
(Charlottetown)
A Romm
Affiliation:
(Toronto)
T Thompson
Affiliation:
(Toronto)
A Piekna
Affiliation:
(Toronto)
Z Wang
Affiliation:
(Toronto)
P Pasceri
Affiliation:
(Toronto)
S Scherer
Affiliation:
(Toronto)
MW Salter
Affiliation:
(Toronto)
J Ellis
Affiliation:
(Toronto)
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Abstract

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Background: Heterozygous loss-of-function mutations in the synaptic scaffolding gene SHANK2 are strongly associated with autism spectrum disorder (ASD). However, their impact on the function of human neurons is unknown. Derivation of induced pluripotent stem cells (iPSC) from affected individuals permits generation of live neurons to answer this question. Methods: We generated iPSCs by reprogramming dermal fibroblasts of neurotypic and ASD-affected donors. To isolate the effect of SHANK2, we used CRISPR/Cas9 to knock out SHANK2 in control iPSCs and correct a heterozygous nonsense mutation in ASD-affected donor iPSCs. We then derived cortical neurons from SOX1+ neural precursor cells differentiated from these iPSCs. Using a novel assay that overcomes line-to-line variability, we compared neuronal morphology, total synapse number, and electrophysiological properties between SHANK2 mutants and controls. Results: Relative to controls, SHANK2 mutant neurons have increased dendrite complexity, dendrite length, total synapse number (1.5-2-fold), and spontaneous excitatory postsynaptic current (sEPSC) frequency (3-7.6-fold). Conclusions: ASD-associated heterozygous loss-of-function mutations in SHANK2 increase synaptic connectivity among human neurons by increasing synapse number and sEPSC frequency. This is partially supported by increased dendrite length and complexity, providing evidence that SHANK2 functions as a suppressor of dendrite branching during neurodevelopment.

Type
POSTER PRESENTATIONS
Copyright
© The Canadian Journal of Neurological Sciences Inc. 2018