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8/16/2019 Pyle Stem Cell Therapy Part 1
1/21
Role of Stem Cells in Muscle
Regeneration and Therapy for
DMD
April Pyle, Ph.D.
Associate Professor
UCLA Department of Microiology, !mmunology
and Molecular "enetics
UCLA #road Stem Cell Research Center
UCLA Center for Duchenne Muscular Dystrophy
8/16/2019 Pyle Stem Cell Therapy Part 1
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Stem Cells$ Properties, Sources %
Rele&ance to DMD
'
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A scientific (ournal )The Stem Cell Chronicle* is
holding a contest for the est ans+er to the
follo+ing uestions on stem cells.
Ta-e a minute and sumit an entry to the contest y
ans+ering the follo+ing$
. AR/ ALL ST/M C/LLS T0/ SAM/1
'. 20AT R/"ULAT/S ST/M C/LL 3AT/1
4ne Minute 2rite
5
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replicate itself
Stem Cell Properties
Self6rene+
2hat is stem cell1
A single cell that can
Differenti
differentiate into many
cell types.
or
7
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OriginAbility to
form othercell types
AbundanceAverage
half-life inculture
/mryonic
8/S, couldform all cell
types9pluripotent
!n &itroartifact
deri&ed from!CM
!mmortal
AdultL!M!T/Dpotential9
multipotent
Rare, tissuespecific Limited
Potency, :umer % Sur&i&al
;
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OriginAbility to
form tumors(teratomas)
Control of differentiation
Clinicaluse
/mryonic 8/SPoorly
understood Acti&e6
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Adult Stem Cells % Sources
? Satellite @muscle
? Myolast @muscle
? Mesangiolast @lood &essels
? Mesenchymal @stromal, one marro+,
fat, etc.
? CD55 positi&e @lood or muscle
? Pericytes @lood &essel or muscle
? Muscle Deri&ed @muscle
? Side Population @lood or muscle
B
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Locating the Satellite Cell in Muscle
? Located et+een the sarcolemma and thee=tracellular matri= of muscle.
? 3ollo+ing in(ury, satellite cells di&ide and gi&e rise
to myolasts.
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Accumulation of Satellite Cells Around the Motor /nd
Plate in an !solated Soleus Myofier
Legend$
Pa=B @green
Satellite cells @arro+s
6ungaroto=in @red
:euromuscular (unction @arro+head
E
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Satellite Cell Myogenesis % Mar-er Progression
Satellite Cell Myofier Myolast
Fuiescent Acti&ated Proliferati&e Commitment todifferentiation
3usion into myotue
Maturation intomyofier
CD57
Pa=B
Myf;
MyoD
Myogenin
MLC53
Molecular Mar-ers /=pressed During Satellite Cell Maturation
G
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Tools for Study and !solation of Satellite Cells
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The Satellite Cell :iche
Signaling +ithin the !mmediate :iche
'
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Harious Cell Types !n&ol&ed in Muscle Regenerat
#lood &esselsIcapillaries
Regenerating myofiers
Macrophages
Satellite cells3irolasts
Angiogenic
endothelial
cells
Differentiating myocytes
3i
pro
5
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Mouse S-eletal Muscle Regeneration
Unin(ured G days 5G da; days
Days Post !n(ury
7
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/mryonic &ersus !nduced Pluripotent Stem Cel
#lastocyst/mryonic Stem Cell @/SC Differentiate
!nduced Pluripotent Stem Cell @iPSC
Reprogramming @adult ac- to pluripotent cell$
4&ere=pression of -ey transcription factors
@e=. 4CT7, JL37, C6M8C, S4K'
/SC 6 deri&ed from inner cell mass of lastocyst iPSC6 deri&ed from reprogram
differentiated cell ac- plurip
;
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Potential Uses of Pluripotent Stem Cells
/mryonic Stem
Cell"enetic Manipulation
gain6of6functionloss6of6function
De&elopmental
#iology
Mesoderm
/ctoderm/ndoderm
Cell Replace
TherapyDrug Disco&ery
>
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0o+ do you Differentiate Pluripotent Stem Cells
Undifferentiated
/S cells
Analye Differentiated Cells
Cells differentiated
as emryonic odies
Cells differentiated
on stromal cells
Cells differentiated
on e=tracellular matri= pr
111!nitiate Differentiation$6L!3, 6emryonic feeders
B
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Patient Specific Approaches for Using DMD6hiPS
4tain patient cells De&elop iPSCs
Transplant corrected muscle cells
De&elop s-e
muscle proge
genetically m
to partially re
functionDe&elop a Pre6Clinical Screening
Tool using Patient Specific Cells
S C ll d D h M l D h
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Stem Cells and Duchenne Muscular Dystrophy
? Dystrophic muscles are highly s
contraction6induced in(ury due t
dystrophin.? This impairs the protecti&e shea
myofiers @the sarcolemma, le
myofier necrosis and acti&atio
satellite cells.? Acti&ated satellite cells di&ide
gi&ing rise to one satellite cell, aproliferating daughter cell that g
myolasts.? The myolasts migrate to the s
differentiate into myocytes, alig
the damaged myofier, repairin
MyofberRegeneration
E
/ h ti f S t llit C ll i DMD
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/=haustion of Satellite Cells in DMD
? 0ea&y urden to repair due to constant
muscle damage
? Lose aility to copy themsel&es
? Lo+ numer affects muscle repair? Damaged muscle fiers are replaced y$
? fat cells @adipose
? scar tissue @firosis
? Muscle can no longer contract effecti&ely
'G
0 C ld St C ll 0 l DMD1
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0o+ Could Stem Cells 0elp DMD1
. Produce ne+ healthy muscle fiers$? "i&e patients normal stem cells to continuously
rene+ muscle
?Correct patients o+n stem cells
ex vivo
andreturn to patient
'. Reduce inflammation or slo+ disease
progression? Secrete )helpful* cyto-ines, proteins, matri=, etc.
5. Personalied genetic medicine
? Screening drugs on patient cells? De&elop patient specific correction strategies
'