Disclosure Paid consultant for Stealth Biotherapeutics David Marcinek Department of Radiology University of Washington dmarc uwedu Muscle Study Group Buffalo NY September 23 2014 ID: 344816
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Slide1
Targeting Mitochondrial Free Radical Production to Improve Aged Skeletal Muscle Function
Disclosure: Paid consultant for Stealth BiotherapeuticsDavid MarcinekDepartment of RadiologyUniversity of Washingtondmarc@uw.edu
Muscle Study Group
Buffalo, NY
September 23, 2014Slide2
Marriotto
; Liao; NIDDK.Slide3
Multifactorial Origins of Sarcopenia
OxidativeDamageReduced Physical ActivityDecreased Anabolic Hormones
Mitochondrial
Dysfunction
Inflammation
Reduced Satellite Cell Function
Neuromuscular
Junction Defects
?Slide4
STRUCTURAL CHANGES
Irreversible (carbonyls, HNE, …)mtDNA mutationsOXIDATIVE DAMAGEMITOCHONDRIAL DEFECTSIntrinsic to mitochondria
MUSCLE DYSFUNCTION
REDOX STRESS
REDOX SIGNALING
Reversible redox modification
Morphology
INHIBITION
Intact system
Homeostasis
Redox signaling
(adaptive)
Redox Stress
(inhibition)
Oxidative Damage
HEALTH
DISEASE
OXIDATIVE STRESS
Marcinek and Siegel. Aging. 2013Slide5
UCP3
H+Redox control of mitochondrial proteins
Mailloux
et al., JBC. 2011
Garcia et al., JBC. 2010
McLain et al., Free
Radic
Res. 2011
Taylor et al., JBC. 2003
Fratelli
et al., PNAS. 2002
Han et al.,
Biochem
. 2005
Odin et al., J
Clin
Invest. 2001
Aon et al.,
Biochim
Biophys
Acta
. 2010Slide6
In Vivo Metabolic Spectroscopy
Siegel et al., (2012) Mech Ageing DevOptical Spectroscopy: O2 cons.
MR Spectroscopy:
ATP prod.
Hindlimb
Optics
Cuff
MR CoilSlide7
ATP
Metabolic Spectroscopy
MbO
2
HbO
2
PCr
CK
Metabolic Spectroscopy
Optical Spectra
31
P NMRSlide8
Parallel effects of ox.
stress and aging on in vivo energetics
#
*
Siegel et al.
Mech
Age
Devel
. 2012
#
*
*Slide9
Aging In Vivo Energetics Similar in Mice and Humans
Conley et al. J Physiol. 2000QUAD
Amara et al. PNAS. 2007
FDISlide10
Summary
Age-related mito deficits similar in mice and elderly humansInduced oxidative stress similar to age-related deficits
Acute
manipulatio
n of oxidative stress will reverse
mito
deficits in aged muscle.
HypothesisSlide11
Szeto
, et al. Antiox and Redox Sig., 2008
Anderson, et al.,
J
Clin
Invest. 2009
Mitochondrial-targeted peptide
Bendavia
(SS-31)
Zhao, et al., JBC. 2004Slide12
Bendavia interacts with cardiolipin
(CL)Birk et al. JASN 2013.CL interacts with each ETC complexNecessary for cristae structureAnchors supercomplexesPreferentially oxidized by stress and disease
MLCL + SS-31
POPC
TLCL
MLCL
SS-31Slide13
H2O
2 ProductionBendavia (SS-31) Mitochondrial H2O2 ProductionSlide14
Bendavia Reverses
In Vivo Mitochondrial DeficitsSiegel et al., Aging Cell. 2013
Inject young and old mice with 3mg/kg SS-31 (~2μM) → in vivo spectroscopy 1 hour later
(5
mo
)
(27
mo
)Slide15
Inject young and old mice with 3mg/kg SS-31 (~2μM) → in vivo spectroscopy 1 hour later
Siegel et al., Aging Cell. 2013Bendavia Reverses In Vivo Mitochondrial DeficitsSlide16
Siegel et al.
, Aging Cell. (2013)Bendavia GSH redox state in aged muscle Slide17
Function?
Does reversal of mitochondrial deficits translate to improved skeletal muscle performance?
Summary
Reduces
mito
H
2
O
2
– not a scavenger.
Acute
Bendavia
reverses
mito
deficits
.
Does not effect healthy
mito
functionSlide18
Improved muscle performance
1 hr after single 3 mg/kgyoung
Siegel et al.,
Aging Cell. 2013 and unpublished
*Slide19
Improved Exercise C
apacity in Old Mice7 daily 3 mg/kg doses
OLD (26
mo
)
YOUNG (6
mo
)Slide20
Improved Exercise C
apacity with 4 week BendaviaMale6 mo25
mo
Female
6
mo
27
moSlide21
Take Home Messages
Mitochondrial redox environment controls in vivo energetics and function in aging skeletal muscle
Bendavia
represents a new class of compound to target mitochondrial oxidative stress.
Acute
Bendavia
improves age-related skeletal muscle
function and exercise tolerance
.Slide22
Phase
IIa test of acute BendaviaPI – Baback Roshanravan
, MD (UW)Coord-Inv – Kevin
Conley, PhD
(UW
)
(SPITM-201)
Randomized, double-blind, placebo-controlled
Single dose, pre, post measurements
Target n=20 per group
Muscle function, in vivo energeticsSlide23
Collaborators
Wei Jun Qian (PNNL)Justin Percival (UW)Terry Kavanagh (UW)Collin White (UW)Dave Brown (ECU)Hazel Szeto (Cornel)
Stealth
Biotherapeutics
Acknowledgements
Marcinek Lab
Michael Siegel (OHSU)
Shane Kruse
Matt Campbell
Rudy
Stuppard
Gary
Knowels
Preston Kendrick
UW Nathan Shock Center
Peter
Rabinovitch
Ann
Chiao
Pabalu
Karunadharma
Nate
Basisty
Mike
MacCoss
Lab Website:
www.blogs.uw.edu
/
marcinekSlide24Slide25
Effect on
Thiol
Redox Status
Sohal
and Orr. Free Rad
Biol
Med. 2012Slide26
Thiol
oxidation: with age, with Bendavia (4wk)
Increased S-glut with age
Partial reversal with SS-31Slide27
Thiol
oxidation: with age, with Bendavia (SS-31, 4wk)Slide28
Take Home Messages
Bendavia improves mitochondrial function and exercise tolerance.
Bendavia
restores redox homeostasis as well as energy homeostasis
.
Restoring redox homeostasis can effect cell physiology through the
thiol
proteome in addition to directly improving energetics. Slide29
NAD
+
H
+
Q
II
H
+
H
+
H
+
H
+
H
+
H
+
H
+
H
+
H
+
H
+
H
+
H
+
H
+
H
+
H
+
H
+
H
+
H
+
H
+
I
III
IV
ADP
P
i
ATP
Proton Motive Force
(predominantly
DY
m
)
H
+
ATP
V
Mitochondrial Matrix
C
Cardiolipin
Peroxidized
Cardiolipin
Reactive Oxygen Species (ROS)
DY
m
ATP
Bendavia
Physiological Mitochondrial Respiration
Pathological Mitochondrial Respiration
OxygenSlide30
Parallel effects of PQ and aging on skeletal muscle energetics
#
*
*
#
*
In Vivo
Maximum
(state 3)
Proton leak
(state 4)
Ex Vivo
p
erm fibers
Siegel et al.
Mech
Age
Devel
. 2012