stem cells: a global perspective...(kondo et al. cell stem cell, 2013) + + background: parkinson’s...
TRANSCRIPT
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Stem Cells: A Global Perspective
Ole Isacson, M.D.–Ph.D.
Harvard Medical School
IMI Stakeholder Forum – 21 May 2014 - Brussels
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Heart and vascular disease
O Isacson
Diabetes
(β-cells)
Aging Processes
(Oxidative stress,…)
Neurological Diseases
(Alzheimer’s…)
Parkinson’s disease (DA cells, etc.)
Regenerative Medicine:
IMI Stakeholder Forum – 21 May 2014 - Brussels
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Regeneration in Planaria – flatworm
IMI Stakeholder Forum – 21 May 2014 - Brussels
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IMI Stakeholder Forum – 21 May 2014 - Brussels
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Regenerative Medicine:
Stem Cell based options
• Mechanism of disease
• Drug screening
• Cell therapy
Study of disease and drug screening
Restorative and cell therapy
IMI Stakeholder Forum – 21 May 2014 - Brussels
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What is a stem cell and an iPS cell?
IMI Stakeholder Forum – 21 May 2014 - Brussels
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Embryonic Stem Cells
IMI Stakeholder Forum- 21 May 2014 - Brussels
http://learn.genetics.utah.edu/content/tech/stemcells/quickref/isolatingEScells.jpg
(1) Sperm and egg join
(2) Embryo develops for 5-7 days
Blastocyst
(3) Remove inner cell mass (4) Grow in dish
(5) Change culture conditions to stimulate cells to differentiate into a variety of cell types
Skin cells Skeletal muscle cells
Neural cells
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Generation of human induced pluripotent stem cells
Davis et al., Trends in Mol Med (2012)
IMI Stakeholder Forum- 21 May 2014 - Brussels
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Why are iPS cells so important?
IMI Stakeholder Forum – 21 May 2014 - Brussels
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Electrophysiological differences between adult human and mouse
Cardiomyocytes
Davis et al., Trends in Mol Med (2012)
IMI Stakeholder Forum- 21 May 2014 - Brussels
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Medicines withdrawn from the market due to cardiotoxicity
Braam et al., Trends in Pharmacological Sciences (2009)
IMI Stakeholder Forum- 21 May 2014 - Brussels
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Human iPS cell derivation, differentiation and applications
Bellin et al., Nature Reviews: Mol Cell Biol (2012)
IMI Stakeholder Forum- 21 May 2014 - Brussels
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The global advancement of medical research & treatment
IMI Stakeholder Forum- 21 May 2014 - Brussels
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Latest developments in stem cell research
IMI Stakeholder Forum – 21 May 2014 - Brussels
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Disease modeling using Alzheimer’s disease patient iPSCs
IMI Stakeholder Forum – 21 May 2014 - Brussels
Patient iPSCs would be useful for precision early diagnosis
Monomer
Dimer
Tetramer
Endosome
ER
ER stress
Oxidative stress ROS↑
BiP↑
PRDX4↑
Amyloidβoligomer
(Kondo et al. Cell Stem Cell, 2013)
+ +
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Background: Parkinson’s disease disease
IMI Stakeholder Forum – 21 May 2014 - Brussels
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VTA/A10
SNc/A9
SNc and VTA relevance to Parkinson’s disease
Isacson, O IMI Stakeholder Forum – 21 May 2014 - Brussels
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The most vulnerable midbrain DA neuron type (A9) in PD -- allowing
basal gangliamovement initiation -- has specific connections,
physiology, cell biology and gene expression
1. Cell type specific gene expression of midbrain dopaminergic neurons.. vulnerability and protection. Chung CY et al Hum. Mol. Genet. 2005
2. Cell type analysis of functional fetal dopamine cell suspension transplants in patients with Parkinson's disease Mendez I et al Brain 2005
IMI Stakeholder Forum – 21 May 2014 - Brussels
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Pre-symptomatic stage
Symptomatic stage
Neuroprotective therapies
Small molecules
Gene therapy
Transplantation and restoration strategies
DA agonists L-DOPA and pharmacology
100%
30%
DA
neu
ron
fu
nct
ion
loss
Isacson, O
Initiation, progression and treatments of midbrain dopaminergic (DA) synaptic and cellular loss in
Parkinson’s disease
Cell Therapy
Age
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Parkinson’s Disease Patients:
Causes and diagnostically defined
human populations
IMI Stakeholder Forum – 21 May 2014 - Brussels
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Pure genetic forms are rare but provide contexts For approaches using PD iPS cells
IMI Stakeholder Forum – 21 May 2014 - Brussels
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Goal: Matching high risk individuals by iPS cell-derived cell function analyses and drug responsiveness
Efficacy
Toxicity
1. 2.
IMI Stakeholder Forum – 21 May 2014 - Brussels
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Strategies for disease modeling in
a dish using iPSCs
Adult cell
Reprogrammed cell (iPSC)
Disease Simulation Process
Readout: Disease outcome/pathology of adaptive responses
1 2 a) none
b) known genetic inherited
a) chemically induced b) genetically induced c) combined a & b
Time
Cell, organelle and molecular phenotypes
Isacson O
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PD patients iPSC derived neural cells: Spontaneous cellular phenotypic changes in dish
IMI Stakeholder Forum – 21 May 2014 - Brussels
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Validation and standardization of individuals’ PD iPS cells
Cooper et al. Science Translational Medicine, 2012
IMI Stakeholder Forum – 21 May 2014 - Brussels
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1. Mitochondrial function and aging
Lysosomal function & autophagy
Immune response
Nuclear transcription response and expression patterns (epigenetic & aging response)
Electrophysiological parameters
3. Transport, cytoskeletal & neurite extension/retraction responses Adaptive electrophysiological response
2. Golgi to Endoplasmic reticulum unfolded protein response and function
Membrane integrity
Dendritic input and neurite morphology
PDiPSC derived neurons can be used to reveal gene or disease specific cell responses
Isacson O
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1. Mitochondrial function and aging
Lysosomal function & autophagy
Immune response
Nuclear transcription response and expression patterns (epigenetic & aging response)
Electrophysiological parameters
Transport, cytoskeletal & neurite responses
Adaptive electrophysiological response
Golgi to Endoplasmic reticulum response and function
Membrane integrity
Dendritic input
Isacson O
PDiPSC derived neurons can be used to reveal gene or disease specific cell responses
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Sanders L et al, 2013
IMI Stakeholder Forum – 21 May 2014 - Brussels
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PD patients iPSC derived neural cells: Cellular phenotypic responses after
challenge in dish
IMI Stakeholder Forum – 21 May 2014 - Brussels
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PINK1 PD iPS cell-derived neuronal cells produce much higher levels of free-radical species after
mitochondrial K+ depolarization than sibling controls
Unstained Healthy Subject Val 1mM
PINK1 Val 1mM
Cooper et al. Science Translational Medicine, 2012
IMI Stakeholder Forum – 21 May 2014 - Brussels
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Genetically linked PD Neural cells from iPS cells are more sensitive to cellular mitochondrial
stress than fibroblasts
Cooper et al. Science Translational Medicine, 2012
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Chemical stressors produce both specific and converging responses from different genetic forms of PD
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Cooper et al. Science Translational Medicine, 2012
Rescue of PD patient neural cell by antioxidant-like and other drugs
IMI Stakeholder Forum – 21 May 2014 - Brussels
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PD patients iPSC derived human
neurons transplanted as an in vivo brain assay
IMI Stakeholder Forum – 21 May 2014 - Brussels
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Transplanted PDiPSC SNCA3x neurons have reduced neurite extension in brain for the
1st month in vivo
IMI Stakeholder Forum – 21 May 2014 - Brussels
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In vitro differentiated then transplanted hPDiPS cells: human specific alpha syn and TH/DA
stained after 6 months in vivo
IMI Stakeholder Forum – 21 May 2014 - Brussels
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Patients iPSC derived cells in a dish: Summary and Outlook
IMI Stakeholder Forum – 21 May 2014 - Brussels
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Strategies for disease modeling in
a dish using iPSCs
Adult cell
Reprogrammed cell (iPSC)
Disease Simulation Process
Readout: Disease outcome/pathology of adaptive responses
1 2 a) none
b) known genetic inherited
a) chemically induced b) genetically induced c) combined a & b
Time
Cell, organelle and molecular phenotypes
Isacson O
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Pre-symptomatic stage
Symptomatic stage
Neuroprotective therapies
Small molecules
Gene therapy
Transplantation and restoration strategies
DA agonists L-DOPA and pharmacology
100%
30%
DA
neu
ron
fu
nct
ion
loss
Isacson, O
Cell Therapy
Outlook: iPSC derived neurons can be used to discover cellular pre-symptomatic or pre-disposing neurobiology and neuroprotective treatments
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Promises and hurdles in using human iPS cells for disease
modeling and drug discovery
• Controls: Human induced pluripotent stem (iPS) cells from the same patient can show biological variability
• Drug discovery and toxicity: Early risk prediction for new drugs reduces attrition rates and costs
• Cell-autonomous versus non-cell-autonomous: Human iPS cells are most suitable for investigating monogenetic diseases with complete penetrance that display cell-autonomous defects on differentiated cells
• Immaturity of differentiated cells: Differentiated cells from human pluripotent stem cells are immature
• Effects of genetic and ethnic backgrounds: Many diseases manifest with incomplete penetrance so that patients harbouring the same mutation have different phenotypes
• Novel biological insights: A key challenge in human iPS cell biology is to generate new (patho)physiological insights.
Bellin et al., Nature Reviews: Mol Cell Biol (2012)
IMI Stakeholder Forum- 21 May 2014 - Brussels
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Penny Hallett Teresia Osborn Oliver Cooper Maria Sundberg Jesse McLean Gaynor Smith Emily Mangano Johan Jansson Tristan Lawson Sarah Izen Eduardo Perez-Torres Jon Beagan Hyemyung Seo Michela Deleidi Gunnar Hargus
Neuroregeneration Laboratory
U Pittsburgh J. Timothy Greenamyre Laurie Sanders Parkinson’s Institute, CA Birgitt Schuele
Harvard University
Kevin Eggan Lee Rubin Paola Arlotta
41
NINDS PDiPS Consortium
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Thank you
IMI Stakeholder Forum – 21 May 2014 - Brussels