Understanding the impact of

Schizophrenia and Autism risk

genes on brain development
Karun K. Singh, Ph.D.
Stem Cell and Cancer Research Institute
Department of Biochemistry and Biomedical Sciences
McMaster University

Genes and
`
Neurodevelopment

ormal Brain Development

Brain Development
Disorder

SCHIZOPHRE
NIA
Genetic
Mutation

AUTISM

Symptoms of`
Schizophrenia

Positive symptoms
Hallucinations
Delusions
Thought disorder (confused thinking and
speech)
Negative symptoms
Diminished emotional expression
Diminished motivation
Inability to experience pleasure
Cognitive symptoms
Poor concentration
Difficult to plan and organize
Poor memory

Treatments
Antipsychotic drugs (old and new
generation)
Cognitive behavior therapy

Neuropathology `of Schizophrenia

N Eng J Med. 1990.

Unaffected twin

Schizophrenic twin

Reduction of brain volume (Grey Matter)

Lateral Ventricle enlargement
Decreased GABAergic interneurons (GABA and GAD67
staining)
Less extensive arborization/neuronal complexity
(dendritic/synapse)

OVERALL Nothing absolutely conclusive

Genetics plays a major role in

`
psychiatric disorders

**Complex genetics at play in

psychiatric disorders**
(Tom Insel, Director NIMH, JCI, 20

2011
Loci for
SZ

Exome Sequencing
Whole Genome Sequencing
more and more genes!

mir137
CSMD1
TRIM26
PCGEM1
Loci for
BPD
ITIH3ITIH4
region

Modeling the` Genetics of

Neurodevelopmental and Psychiatric
Disorders
Brain DiseasePatient Cells
Risk Gene

Developing
Mouse Brain
Why do this?

Human Cellular
Reprogramming

1. Understand how genes impact brain structure/function during development

2. Understand core signaling pathways affected

Disrupted in Schizophrenia-1 (DISC1)

Chromosome
Chromosome (1;
(1; 11)
11) translocation
translocation
disrupts
disrupts the
the DISC1
DISC1 locus.
locus.

Experimental model
to
examine
`
DISC1/Dixdc1: The developing
mouse
cortex to study
Approach: In utero
electroporation
neural stem cells and neuronal
differentiation/maturation

CP Direction of
migration

neurons

IZ

axon tracts

VZ

neural stem

Dixdc1 functionally
interacts with
`
DISC1 to regulate neural progenitor
proliferation

Singh et al., 201

Dixdc1 and DISC1 `regulate neuronal

migration

DIS
Frag2 DIXDC1
C1

Singh et al., 2010

Dixdc1 is a critical
regulator of
`
DISC1 and embryonic brain
development

Early brain development

Mid-brain development

Singh et al., 201

Does Dixdc1 functionally

interact
`
with DISC1 in neural connectivity
development?

Dendrite Growth
Spine Structure
Synapse Formation

Penzes et al., Nat Neurosci 2011

Does a DISC1-Dixdc1
pathway
`
regulate the growth of dendrites
and synapses?
Control shRNA DISC1 shRNA
Dixdc1 shRNA
Dixdc1 Frag2

DIV6
Total Dendrite Branch Length

Sholl Analysis
Ctrl shRNA
Disc1 shRNA
Dixdc1 shRNA
Dix Frag2

Vickie Kwan, preliminary resu

Confirming this in vivo in utero electroporation

Examining synapses with genetic

`
tools:
Trans-synaptic
labeling using Rabies virus
Advantages
1. Single Cell
Resolution
2. Watch
dynamics
over time

Ed Callaway Lab
Salk Institute

`
Trans-synaptic labeling
in mouse
DIV 10 neurons
DIV 14
IUE E15 Culturecortical
E17

LV-Syn-HTG

Infect: Fix
Rabies cells

How does DISC/Dixdc1 regulate

synapse formation?

Rabies virus courtesy of

Ian Wickersham and
Heather Sullivan (Seung

Studying the` Genetics of

Autism

Not included:

Additional CNVs
Exome
sequencing
(de novo
mutations)
Whole genome
sequencing
Aldinger et al., 2011 Neuro

Human cellular
`
reprogramming
to create
patient-derived neural cells
Advantages
Model autism
genes
Behavior assays
Neural circuits
and cell types
involved

Some Disadvantages
Difficult to model
disease gene networks,
large CNVs, human
SNPs
Gene disruption doesnt
completely mimic
human genetics (eg.
KOmutation)
Not patient brain tissue
(human brain specific)

Human cellular
`
reprogramming
to create
patient-derived neural cells
ing
m
am
r
g
ro ne s
p
e
R
ge

D if
ere
n

tiat
e

Induced
pluripotent
stem (iPS) cells

Direct conversion
Induced Neuronal (iN) or
PatientNeural Progenitor (iNP) cells derived

patient
skin
samples
NEURAL
ADVANTAGES of Direct Conversion:
CELLS
1. Faster than iPS method
2. Epigenetic signature of patient cell is likely preserved
3. Specific Neuronal subtype generation (Spinal Motor, Dopaminergic)

Studying the` Genetics of

Autism

Aldinger et al., 2011 Neuro

Tuberous Sclerosis Complex (TSC): a

genetic disorder` with high rates of
autism

Collaboration with Dr. Philippe Major, Sainte-Justine

Hospital, Montreal
1. Benign tumors in vital organs including brain
2. Autism features (syndromic autism) 25-60% in ASD
3. Learning disabilities, developmental delay
4. Epilepsy

Mouse
Models
Protein
Translatio
n
Plasticity
(mGluRLTD)

Kelleher, III and Bear, 2012 C

Healthy or
TSC
Fibroblasts

Human
Neural
Cells

P
A h en
ss o
ay ty
D pe
ev a
el n d
op
m
en
t

D
R ire
ep ct
ro C
gr ell
am ul
m ar
in
g

Using Cellular Reprogramming to Study

`
TSC and Autism

Synapse Function
Dendrite/Spine Growth
Trans-synaptic labeling
Automated Electrophysiology

Drug
Screen

In vivo Human Neuronal Model

`
using
Xenotransplantation
Patientderived
Fluorescent LabelTransplant
neural cells

o profile of human cells:

w do patient neural cells functionally integrate into the developi

adult brain?

Acknowledgements
My lab:
Vickie Kwan
Shashwat Desai
Omar Shehab

University of
MontrealSainte Justine
Hospital
Dr. Philippe Major
(TSC)

Massachusetts
Institute of
Technology:
Dr. Li-Huei Tsai

Funding:
Ontario Research
Fund