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