neuroprotective and regenerative stem cell therapies in a pig … · 2020-02-19 · 1) a...
TRANSCRIPT
![Page 1: Neuroprotective and Regenerative Stem Cell Therapies in a Pig … · 2020-02-19 · 1) a regenerative cell therapy that will not only protect cells from ischemic injury but replace](https://reader033.vdocuments.net/reader033/viewer/2022060500/5f1a6381e75dad1bf51b91f7/html5/thumbnails/1.jpg)
Neuroprotective and Regenerative Stem Cell
Therapies in a Pig Stroke Model
Franklin D. WestRegenerative Bioscience Center
University of Georgia
![Page 2: Neuroprotective and Regenerative Stem Cell Therapies in a Pig … · 2020-02-19 · 1) a regenerative cell therapy that will not only protect cells from ischemic injury but replace](https://reader033.vdocuments.net/reader033/viewer/2022060500/5f1a6381e75dad1bf51b91f7/html5/thumbnails/2.jpg)
Financial Disclosure
• No financials to disclose
![Page 3: Neuroprotective and Regenerative Stem Cell Therapies in a Pig … · 2020-02-19 · 1) a regenerative cell therapy that will not only protect cells from ischemic injury but replace](https://reader033.vdocuments.net/reader033/viewer/2022060500/5f1a6381e75dad1bf51b91f7/html5/thumbnails/3.jpg)
• Second leading cause of death globally
• Number one cause of long term disability in US
• Every 40 seconds someone has a stroke
• Every 4 minutes someone dies of stroke
Mozaffarian et al. Circulation. 2015.
www.educatehealth.ca
Stroke: A Global Killer
![Page 4: Neuroprotective and Regenerative Stem Cell Therapies in a Pig … · 2020-02-19 · 1) a regenerative cell therapy that will not only protect cells from ischemic injury but replace](https://reader033.vdocuments.net/reader033/viewer/2022060500/5f1a6381e75dad1bf51b91f7/html5/thumbnails/4.jpg)
![Page 5: Neuroprotective and Regenerative Stem Cell Therapies in a Pig … · 2020-02-19 · 1) a regenerative cell therapy that will not only protect cells from ischemic injury but replace](https://reader033.vdocuments.net/reader033/viewer/2022060500/5f1a6381e75dad1bf51b91f7/html5/thumbnails/5.jpg)
Over 700 treatments have gone to clinical trial for stroke and only two have been FDA approved:
▪Tissue Plasminogen Activator (tPA)▪Thrombectomy
•Limited window of treatment•No regenerative potential•<25% of patients can receive
tPA and/or thrombectomy
Peter McMeekin, et al. European Stroke Journal, 2017. 2(4): p. 319-326; Sharma, V.K., et al. Stroke Res Treat, 2010; Lapchak, P.A., Transl Stroke Res, 2010; Serbest, G., et al. FASEB J, 2006.
Limited Treatments for Stroke
![Page 6: Neuroprotective and Regenerative Stem Cell Therapies in a Pig … · 2020-02-19 · 1) a regenerative cell therapy that will not only protect cells from ischemic injury but replace](https://reader033.vdocuments.net/reader033/viewer/2022060500/5f1a6381e75dad1bf51b91f7/html5/thumbnails/6.jpg)
An assessment of failed treatments by the Stem Cell Therapies as an Emerging Paradigm in Stroke (STEPS) group has resulted in the identifying 2 major needs of the research community:
1) a regenerative cell therapy that will not only protect cells from ischemic injury but replace the lost and damaged tissues
2) a translational animal model more similar to humans for testing treatments
Stem Cell Therapies as an Emerging Paradigm in Stroke (STEPS): bridging basic and clinical science for cellular and neurogenic factor therapy in treating stroke. Stroke, 2009. Savitz, S.I., et al., Stem Cell Therapy as an Emerging Paradigm for Stroke (STEPS) II. Stroke, 2011.
Why Do All the Drugs Fail?
![Page 7: Neuroprotective and Regenerative Stem Cell Therapies in a Pig … · 2020-02-19 · 1) a regenerative cell therapy that will not only protect cells from ischemic injury but replace](https://reader033.vdocuments.net/reader033/viewer/2022060500/5f1a6381e75dad1bf51b91f7/html5/thumbnails/7.jpg)
Induced Pluripotent Stem Cell-Derived Neural Stem Cells (iNSC) to Treat Neural Injury
![Page 8: Neuroprotective and Regenerative Stem Cell Therapies in a Pig … · 2020-02-19 · 1) a regenerative cell therapy that will not only protect cells from ischemic injury but replace](https://reader033.vdocuments.net/reader033/viewer/2022060500/5f1a6381e75dad1bf51b91f7/html5/thumbnails/8.jpg)
Induced Pluripotent Stem Cell-Derived Neural Stem Cells (iNSC) to Treat Neural Injury
![Page 9: Neuroprotective and Regenerative Stem Cell Therapies in a Pig … · 2020-02-19 · 1) a regenerative cell therapy that will not only protect cells from ischemic injury but replace](https://reader033.vdocuments.net/reader033/viewer/2022060500/5f1a6381e75dad1bf51b91f7/html5/thumbnails/9.jpg)
iPSC-NSCs Express SOX1 and Nestin
SOX1 Nestin
Merge
![Page 10: Neuroprotective and Regenerative Stem Cell Therapies in a Pig … · 2020-02-19 · 1) a regenerative cell therapy that will not only protect cells from ischemic injury but replace](https://reader033.vdocuments.net/reader033/viewer/2022060500/5f1a6381e75dad1bf51b91f7/html5/thumbnails/10.jpg)
Differentiated iPSC-NSCs Express βIII-Tubulin and MAP2 Neuron Markers
βIII Tubulin MAP2
Merge
![Page 11: Neuroprotective and Regenerative Stem Cell Therapies in a Pig … · 2020-02-19 · 1) a regenerative cell therapy that will not only protect cells from ischemic injury but replace](https://reader033.vdocuments.net/reader033/viewer/2022060500/5f1a6381e75dad1bf51b91f7/html5/thumbnails/11.jpg)
Differentiated iPSC-NSCs Express the GFAP Astrocyte Marker
GFAP Merge
![Page 12: Neuroprotective and Regenerative Stem Cell Therapies in a Pig … · 2020-02-19 · 1) a regenerative cell therapy that will not only protect cells from ischemic injury but replace](https://reader033.vdocuments.net/reader033/viewer/2022060500/5f1a6381e75dad1bf51b91f7/html5/thumbnails/12.jpg)
Differentiated iPSC-NSCs Express the O4 Oligodendrocyte Marker
MergeO4
![Page 13: Neuroprotective and Regenerative Stem Cell Therapies in a Pig … · 2020-02-19 · 1) a regenerative cell therapy that will not only protect cells from ischemic injury but replace](https://reader033.vdocuments.net/reader033/viewer/2022060500/5f1a6381e75dad1bf51b91f7/html5/thumbnails/13.jpg)
An assessment of failed treatments by the Stem Cell Therapies as an Emerging Paradigm in Stroke (STEPS) group has resulted in the identifying 2 major needs of the research community:
1) a regenerative cell therapy that will not only protect cells from ischemic injury but replace the lost and damaged tissues
2) a translational animal model more similar to humans for testing treatments
Stem Cell Therapies as an Emerging Paradigm in Stroke (STEPS): bridging basic and clinical science for cellular and neurogenic factor therapy in treating stroke. Stroke, 2009. Savitz, S.I., et al., Stem Cell Therapy as an Emerging Paradigm for Stroke (STEPS) II. Stroke, 2011.
Why Do All the Drugs Fail?
![Page 14: Neuroprotective and Regenerative Stem Cell Therapies in a Pig … · 2020-02-19 · 1) a regenerative cell therapy that will not only protect cells from ischemic injury but replace](https://reader033.vdocuments.net/reader033/viewer/2022060500/5f1a6381e75dad1bf51b91f7/html5/thumbnails/14.jpg)
1.Size
2.Lissencephalic Vs Gyrencephalic
3.White Matter
Stem Cell Therapies as an Emerging Paradigm in Stroke (STEPS): bridging basic and clinical science for cellular and neurogenic factor therapy in treating stroke. Stroke, 2009; Nakamura, M., et al. J Neurosurg Pediatr, 2009; Kuluz, J.W., et al. Stroke, 2007; Tanaka, Y., et al. Stroke, 2008; Recommendations for standards regarding preclinical neuroprotective and restorative drug development. Stroke; DeFelipe, J. Frontiers in Neuroanatomy, 2011.
Differences in the Rodent Brain Relative to Human Brain
![Page 15: Neuroprotective and Regenerative Stem Cell Therapies in a Pig … · 2020-02-19 · 1) a regenerative cell therapy that will not only protect cells from ischemic injury but replace](https://reader033.vdocuments.net/reader033/viewer/2022060500/5f1a6381e75dad1bf51b91f7/html5/thumbnails/15.jpg)
![Page 16: Neuroprotective and Regenerative Stem Cell Therapies in a Pig … · 2020-02-19 · 1) a regenerative cell therapy that will not only protect cells from ischemic injury but replace](https://reader033.vdocuments.net/reader033/viewer/2022060500/5f1a6381e75dad1bf51b91f7/html5/thumbnails/16.jpg)
T2-FLAIR DWI ADC
![Page 17: Neuroprotective and Regenerative Stem Cell Therapies in a Pig … · 2020-02-19 · 1) a regenerative cell therapy that will not only protect cells from ischemic injury but replace](https://reader033.vdocuments.net/reader033/viewer/2022060500/5f1a6381e75dad1bf51b91f7/html5/thumbnails/17.jpg)
An assessment of failed treatments by the Stem Cell Therapies as an Emerging Paradigm in Stroke (STEPS) group has resulted in the identifying 2 major needs of the research community:
1) a regenerative cell therapy that will not only protect cells from ischemic injury but replace the lost and damaged tissues
2) a translational animal model more similar to humans for testing treatments
Stem Cell Therapies as an Emerging Paradigm in Stroke (STEPS): bridging basic and clinical science for cellular and neurogenic factor therapy in treating stroke. Stroke, 2009. Savitz, S.I., et al., Stem Cell Therapy as an Emerging Paradigm for Stroke (STEPS) II. Stroke, 2011.
Why Do All the Drugs Fail?
![Page 18: Neuroprotective and Regenerative Stem Cell Therapies in a Pig … · 2020-02-19 · 1) a regenerative cell therapy that will not only protect cells from ischemic injury but replace](https://reader033.vdocuments.net/reader033/viewer/2022060500/5f1a6381e75dad1bf51b91f7/html5/thumbnails/18.jpg)
Overarching Hypothesis
Transplanted iNSCs will produce regenerative
trophic factors and undergo differentiation
leading to cellular, tissue and functional
recovery in a pig model of ischemic stroke.
Emily Baker
![Page 19: Neuroprotective and Regenerative Stem Cell Therapies in a Pig … · 2020-02-19 · 1) a regenerative cell therapy that will not only protect cells from ischemic injury but replace](https://reader033.vdocuments.net/reader033/viewer/2022060500/5f1a6381e75dad1bf51b91f7/html5/thumbnails/19.jpg)
Experimental Design
Tissue Collection
Sample Collection:
1. Magnetic Resonance Imaging (MRI)2. Immunohistochemistry
MRI
10x106
![Page 20: Neuroprotective and Regenerative Stem Cell Therapies in a Pig … · 2020-02-19 · 1) a regenerative cell therapy that will not only protect cells from ischemic injury but replace](https://reader033.vdocuments.net/reader033/viewer/2022060500/5f1a6381e75dad1bf51b91f7/html5/thumbnails/20.jpg)
DWI and ADC Maps Confirm Ischemic Stroke
T2-FLAIR DWI ADC
![Page 21: Neuroprotective and Regenerative Stem Cell Therapies in a Pig … · 2020-02-19 · 1) a regenerative cell therapy that will not only protect cells from ischemic injury but replace](https://reader033.vdocuments.net/reader033/viewer/2022060500/5f1a6381e75dad1bf51b91f7/html5/thumbnails/21.jpg)
iNSC Treatment Leads to Slowed Tissue Atrophy
Tis
su
e V
olu
me
Ch
an
ge
(% o
f C
on
tra
late
ral
He
mis
ph
ere
)
24hr 1wk 4wk 12wk-20
-10
0
10
20 Non-TreatediNSC Treated
***** *
*#
24hr 1wk
4wk 12wk
T2 FLAIR
indicates significance from contralateral hemisphere# indicates significance between treatment groups
Normal
![Page 22: Neuroprotective and Regenerative Stem Cell Therapies in a Pig … · 2020-02-19 · 1) a regenerative cell therapy that will not only protect cells from ischemic injury but replace](https://reader033.vdocuments.net/reader033/viewer/2022060500/5f1a6381e75dad1bf51b91f7/html5/thumbnails/22.jpg)
iNSC Treatment Leads to Improved White Matter Integrity
indicates significance from 24 hours, within treatmentiN
SC
Tre
ate
dN
on
-Tre
ate
d
Ch
an
ge in
FA
24hr 1wk 4wk 12wk0
50
100
150
200
250
300
350
400 Non-Treated
iNSC Treated
*Normal
![Page 23: Neuroprotective and Regenerative Stem Cell Therapies in a Pig … · 2020-02-19 · 1) a regenerative cell therapy that will not only protect cells from ischemic injury but replace](https://reader033.vdocuments.net/reader033/viewer/2022060500/5f1a6381e75dad1bf51b91f7/html5/thumbnails/23.jpg)
iNSC Treatment Leads to Recovery in NAA C
han
ge in
NA
A
24hr 1wk 4wk 12wk0
2
4
6
8
10Non-Treated
iNSC Treated
*#
#
*
No
n-T
reat
ed
iNSC
Tre
ate
dindicates significance between treatment groups
# indicates significance from 24 hours, within treatment
Normal
![Page 24: Neuroprotective and Regenerative Stem Cell Therapies in a Pig … · 2020-02-19 · 1) a regenerative cell therapy that will not only protect cells from ischemic injury but replace](https://reader033.vdocuments.net/reader033/viewer/2022060500/5f1a6381e75dad1bf51b91f7/html5/thumbnails/24.jpg)
iNSC Treatment Leads to Recovery in Choline
Ch
an
ge in
Ch
o
24hr 1wk 4wk 12wk0
2
4
6
8
10Non-Treated
iNSC Treated
* *
No
n-T
reat
ed
iNSC
Tre
ate
dindicates significance between treatment groups
Normal
![Page 25: Neuroprotective and Regenerative Stem Cell Therapies in a Pig … · 2020-02-19 · 1) a regenerative cell therapy that will not only protect cells from ischemic injury but replace](https://reader033.vdocuments.net/reader033/viewer/2022060500/5f1a6381e75dad1bf51b91f7/html5/thumbnails/25.jpg)
iNSC Treatment Leads to Recovery in Creatine
Ch
an
ge in
Cr
24hr 1wk 4wk 12wk0
5
10
15Non-Treated
iNSC Treated
*
No
n-T
reat
ed
iNSC
Tre
ate
dindicates significance between treatment groups
Normal
![Page 26: Neuroprotective and Regenerative Stem Cell Therapies in a Pig … · 2020-02-19 · 1) a regenerative cell therapy that will not only protect cells from ischemic injury but replace](https://reader033.vdocuments.net/reader033/viewer/2022060500/5f1a6381e75dad1bf51b91f7/html5/thumbnails/26.jpg)
iNSC Treatment Improves Cerebral Blood Perfusion
% o
f C
on
tra
late
ral
He
mis
ph
ere
24hr 1wk 4wk 12wk0
50
100
150
200 Non-TreatediNSC Treated
*
Mean Transit Time%
of
Co
ntr
ala
tera
l H
em
isp
he
re
24hr 1wk 4wk 12wk0
50
100
150
200 Non-TreatediNSC Treated
*
Time to Peak
iNSC
Tre
ate
dN
on
-Tre
ate
d
indicates significance between treatment groups
Normal
Normal
![Page 27: Neuroprotective and Regenerative Stem Cell Therapies in a Pig … · 2020-02-19 · 1) a regenerative cell therapy that will not only protect cells from ischemic injury but replace](https://reader033.vdocuments.net/reader033/viewer/2022060500/5f1a6381e75dad1bf51b91f7/html5/thumbnails/27.jpg)
iNSC Treatment Reduces Neuron Loss at the Lesion Border
A
DNormal iNSC TreatedNon-Treated
NeuNNeuN CBA
Ne
uN
B
Neu
N+
cel
ls/m
m2
Normal Non-Treated iNSC Treated0
50
100
150
200
250
300
350
*
D
![Page 28: Neuroprotective and Regenerative Stem Cell Therapies in a Pig … · 2020-02-19 · 1) a regenerative cell therapy that will not only protect cells from ischemic injury but replace](https://reader033.vdocuments.net/reader033/viewer/2022060500/5f1a6381e75dad1bf51b91f7/html5/thumbnails/28.jpg)
iNSC Treatment Reduces Microglia Activation at the Lesion Border
A
% Ib
a1+
Are
a
Normal Non-Treated iNSC Treated0
5
10
15
20
25 # #
#
H
F G HE
Iba1
F GNormal iNSC TreatedNon-Treated
![Page 29: Neuroprotective and Regenerative Stem Cell Therapies in a Pig … · 2020-02-19 · 1) a regenerative cell therapy that will not only protect cells from ischemic injury but replace](https://reader033.vdocuments.net/reader033/viewer/2022060500/5f1a6381e75dad1bf51b91f7/html5/thumbnails/29.jpg)
iNSC Treatment Promotes Endogenous NeuroblastProliferation and Migration to the Lesion Border
DC
X+
cells
/mm
2
Normal Non-Treated iNSC Treated0
5
10
15
20
***
D
A B CNormal Non-Treated iNSC Treated
DC
X
![Page 30: Neuroprotective and Regenerative Stem Cell Therapies in a Pig … · 2020-02-19 · 1) a regenerative cell therapy that will not only protect cells from ischemic injury but replace](https://reader033.vdocuments.net/reader033/viewer/2022060500/5f1a6381e75dad1bf51b91f7/html5/thumbnails/30.jpg)
Transplanted iNSCs Differentiate into NeuronsHNA
NeuN HNA NeuN DAPI
![Page 31: Neuroprotective and Regenerative Stem Cell Therapies in a Pig … · 2020-02-19 · 1) a regenerative cell therapy that will not only protect cells from ischemic injury but replace](https://reader033.vdocuments.net/reader033/viewer/2022060500/5f1a6381e75dad1bf51b91f7/html5/thumbnails/31.jpg)
Transplanted iNSCs Differentiate into Neurons
HNA NeuN HNA NeuN DAPI
![Page 32: Neuroprotective and Regenerative Stem Cell Therapies in a Pig … · 2020-02-19 · 1) a regenerative cell therapy that will not only protect cells from ischemic injury but replace](https://reader033.vdocuments.net/reader033/viewer/2022060500/5f1a6381e75dad1bf51b91f7/html5/thumbnails/32.jpg)
Transplanted iNSCs Differentiate into Oligodendrocytes
HNA Olig2 HNA Olig2 DAPI
![Page 33: Neuroprotective and Regenerative Stem Cell Therapies in a Pig … · 2020-02-19 · 1) a regenerative cell therapy that will not only protect cells from ischemic injury but replace](https://reader033.vdocuments.net/reader033/viewer/2022060500/5f1a6381e75dad1bf51b91f7/html5/thumbnails/33.jpg)
Transplanted iNSCs Differentiate Primarily into Neurons
% C
olo
calizati
on
wit
h H
NA
NeuN Olig20
20
40
60
80
100
![Page 34: Neuroprotective and Regenerative Stem Cell Therapies in a Pig … · 2020-02-19 · 1) a regenerative cell therapy that will not only protect cells from ischemic injury but replace](https://reader033.vdocuments.net/reader033/viewer/2022060500/5f1a6381e75dad1bf51b91f7/html5/thumbnails/34.jpg)
ConclusionsiNSC treatment promotes brain tissue recovery through:
– Improved white matter integrity
– Improved neurometabolism
– Enhanced cerebral perfusion
– Neuroprotective
– Promoted neurogenesis
– Cell replacement
– Immunomodulatory
![Page 35: Neuroprotective and Regenerative Stem Cell Therapies in a Pig … · 2020-02-19 · 1) a regenerative cell therapy that will not only protect cells from ischemic injury but replace](https://reader033.vdocuments.net/reader033/viewer/2022060500/5f1a6381e75dad1bf51b91f7/html5/thumbnails/35.jpg)
Funding Sources:NIH-NINDS, DoD, Bill and Melinda Gates Foundation, USDA, ArunA Biomedical, REM, BIRC
Collaborators:Simon Platt, DVMElizabeth Howerth, DVM/PhDKylee Jo Duberstein, PhDSteve Stice, PhDDavid Hess, MDQun Zhao, PhDHea Jin Park, PhD
West Laboratory:Emily Baker, PhDVivian Lau, DVM,MSHarrison Grace, PhDHolly Kinder, PhDErin KaiserKelly ScheulinElizabeth WatersSoo Shin Sydney Sneed