Using brain organoids to understand neurological diseases · density of RGCs (asterisk) and...
Transcript of Using brain organoids to understand neurological diseases · density of RGCs (asterisk) and...
Using brain organoids to understand neurological diseases
Stem cells and organoids:
Unlocking their potential to
understand and treat disease
02082018
Guo-li Ming
Department of Neuroscience
University of Pennsylvania
Models to study human biology and disorders
(practicallysceince.com)
Difference between human and rodent cortical development
Mouse
Outer
Subventricular
zone (oSVZ)
Human • Stem cell pool
• Neuronal diversity
• Complex circuitry
Miniaturized multi-well spin bioreactor SpinΩ
standard
bioreactors
miniaturized
bioreactors
3D designed & printed miniaturized bioreactors
(by three high schoolers)
Miniaturized multi-well spin bioreactor SpinΩ
standard
bioreactors
miniaturized
bioreactors
3D designed & printed miniaturized bioreactors
(by three high schoolers)
Brain region-specific forebrain organoids –Laminar organization in forebrain organoids recapitulate
human cortical neurogenesis
Day 1
Neural Induction
Day 4 Day 7
Forebrain Patterning
Day 13
Differentiation media
Matrigel SpinΩ
Day 71
Maturation media
Day 14
Forebrain organoids as an experimental model system
• Laminar organization with all
6 cortical layers
Spontaneous synaptic
currents
Glutamatergic GABAergic
Dorsolateral
prefrontal
cortex
• Human specific oSVZ neural
stem cells
• Functional neurons
• Resembling fetal cortical
brain development up to the
end of second trimester at
transcriptional level
Suspected link of microcephaly cases to Zika virus outbreak
(from Driggers et al. N Engl J Med 2016)
ZIKV infection in early stage forebrain organoids causes devastating damage
Day 14+10
ZIK
VE
SO
X2 D
AP
I
GW4 GW8 Conception
Neurulation Neurogenesis
Organoid
Pregnancy
Modeling prenatal ZIKV exposure using forebrain organoids
• Increase cell
• death
• Target neural
stem cells
• Decrease cell
proliferation
ZIKV infection causes defects resembling microcephaly
(Qian* and Nguyen* et al. Cell 2016)
Therapeutics: antiviral & neuroprotective drug screens
A collection of 6000 compounds:
• FDA approved drugs
• Clinical trial drug candidates
• Pharmacological active
Wei Zheng
at National Center for Advancing
Translational Sciences (NCATs)
Therapeutics: antiviral & neuroprotective drug screens
Miao et al. Nature Medicine 2016)
Neurogenesis and disease modeling using hiPSCs
• Biology of human brain
development
• Mechanisms of
neurodevelopmental
and psychiatric
disorders
hippocampus
Acknowledgements
NINDS, NIMH, MSCRF, SAFRI
Hongjun Song @ Upenn
Xuyu Qian (BME)
Ha Nam Nguyen (CEO, 3Dnamics)
Fadi Jacob (Neuroscience)
Zhexing Wen (Emory)
Ki-jun Yoon
Wei-kai Huang (Pathobiology)
High school students
Max Song
Chris Hadiono
William Jeang
WSE Machine Shop@JHU
Niel Leon
Daren Ayres
@JHU
Hai-quan Mao (Material Sciences)
Heng Zhu (Pharmacology)
Jiang Qian (Ophthalmology)
Andrew Holland (Cell Biology)
Gabsang Lee (Neurology)
Fred Gage
Abed Al Fattah Mansour
Hengli Tang @
Florida State University
Christy Hammack
Sarah C. Ogden
Emily M. Lee
Yichen Cheng
Peng Jin @ Emory
Bing Yao
Feiran Zhang
Hao Wu
Brady Maher @
Lieber Institute
Gregory Hamersky
Wei Zheng @ NCATs
A mini-workshop on production of mini-bioreactors
and region-specific brain organoids (June 17, 2016)
Lancaster et al.
Nature (2013)
Kadoshima et al.
PNAS (2013)
Differentiation of hESC or hiPSCs into brain organoids in 3D
TUJ1 SOX2 Hoechst
Low
Attachment
dishes
Spinning
bioreactor
Cerebral
organoids
Cell growth and migration in forebrain organoids
Towards making a six-layer human cortex in organoids
Functional glutamatergic and GABAergic neurons
Day 100 Forebrain organoids
Spontaneous synaptic currents
Glutamatergic GABAergic
GFPSV2
Brady Maher at Lieber Institute
GABAergic neuron subtypes astrocytes
Disease modeling using
hiPSC-based Organoids
CTIP2 PKC-l DAPI
Zika virus (ZIKV) infects hiPSC-derived neural progenitor cells in 2D culture
(Tang et al. Cell Stem Cell. 2016;
Zheng et al. Nucleic Acid Research 2016)
• Productive
infection
• Increased cell
death
• Decreased cell
proliferation
• Down-regulation
of cell cycle genes
Hengli Tang at Florida State University
Peng Jin at Emory University
Understanding mechanisms underlying ZIKV pathogenesis
Understanding mechanisms underlying ZIKV pathogenesis
GFP
GF
P E
dU
DA
PI
GFP
Electroporation of forebrain organoids
EdU/DAPI
NS2A GFP
ZIKV-NS2A disrupts adherens junction complex formation in forebrain organoids
Yoon et al. Cell Stem Cell 2017
ZIKV-NS2A expression leads to disorganized cortical structure in forebrain organoids
“Periventricular region showing dysplastic intraventricular protrusions with lower density of RGCs (asterisk) and disorganized RG morphology and processes” Onorati et al., Cell Report 2016
Yoon et al. Cell Stem Cell 2017