Download MMIG 6/9/2010

Welcome to MMIG
(Mitochondria and Metabolism
Interest Group)
What is MMIG?
• Interest-driven group open to all PIs
studying M & M, currently ~25 members
•
•
•
•
Scientific exchange and collaboration
Share resources and expertise
Strengthen training environment
Program development and visibility
Format
• Regular meetings: weekly, biweekly or
monthly?
• Invited speakers/visitors (local and
national): Monthly?
• Forum for the trainees (journal club and
work-in-progress presentations)
• Annual retreat: November 8-9, 2010
• Home base: Mitochondria and Metabolism
Center at SLU
http://depts.washington.edu/mmcslu
• Program Specialist: Karen Liebert
[email protected]
– Refer new MMIG members
– Suggest future meeting place
– Suggest speakers, meeting time and frequency
Mitochondrial
Mutants
gas-1
clk-1
mev-1
isp-1
ctb-1
MMIG - 9 June 2010 - Morgan/Sedensky 1
Affected complex in kids?
EC50s for BIS=60
% sevoflurane
5
4
complex I
3
non complex I
2
normal
1
0
Mitochondrial Defects
MMIG - 9 June 2010 - Morgan/Sedensky 2
Respiratory Supercomplexes
in C. elegans
MMIG - 9 June 2010 - Morgan/Sedensky 3
(Suthammarak et al, JBC 2008)
The complexities of regulating mitochondria in neurons
How is mitochondrial dynamics (fission/fusion) regulated in neurons?
Is the fission/fusion machinery in neurons unique?
How is mitochondrial fission/fusion regulated in neurons in response
to injury and disease?
Can we prevent neuronal apoptosis and maintain function by
modulating mitochondrial dynamics?
MMIG - 9 June 2010 - Morrison 1
MMIG - 9 June 2010 - Morrison 2
MMIG - 9 June 2010 - Morrison 3
Wang lab Research interests:
 Physiological mitochondrial permeability transition pore (mPTP) opening
and superoxide flash activity in the heart;
 Molecular mechanism of mPTP triggered superoxide flash production;
 Role of mitochondrial reactive oxygen species (ROS) in oxidative stress.
What we do?
 Focus on cardiovascular system;
 Adult cardiac myocyte culture and adenovirus mediated gene transfer;
 Confocal measurement of superoxide flash (cpYFP), mitochondrial /
cytosolic Ca2+ and mitochondrial membrane potential;
 Assessment of cardiac myocyte function: contractility & Ca2+ handling;
 Transgenic mice expressing superoxide indicator in mitochondria;
 Mitochondrial ROS signaling in disease: ischemia reperfusion injury,
heart failure, diabetes and metabolic syndrome.
MMIG - 9 June 2010 - Wang 1
Single Mitochondrial Superoxide Flash in Adult Cardiac Myocyte
10 m
Time (s)
488 nm
TMRM
488 nm
488 nm
405 nm
405 nm
10 s
20 s
MMIG - 9 June 2010 - Wang 2
0.5 F/F0
0.3 F/F0
405 nm
(Wang et al, 2008, Cell 134:279-290)
Integrative Study of Mitochondrial Function in Health & Disease
MMIG - 9 June 2010 - Wang 3
https://depts.washington.edu/mmcslu/home
PLC
How do mitochondria produce ROS?
How are mitochondrial ROS ‘decoded’?
Apoptosis
PKC
ym loss
TCA
ROS
Bioenergetics
NF-kB
Inflammation
MMIG - 9 June 2010 - Hawkins 1
DAG
IP3
Relationship between cytosolic calcium and
mitochondrial ROS ROS
MMIG - 9 June 2010 - Hawkins 2
Laboratory Interests
•
How do disparate mitochondrial calcium uptake patterns translate to cellular
function?
Microvascular
EC
Macrovascular
EC
•
How do calcium-linked oxidants influence mitochondrial function?
•
What are the cellular targets of mitochondrial reactive oxygen species?
MMIG - 9 June 2010 - Hawkins 3
Mitochondrial hub in apoptosis
Smart bombs
MMIG - 9 June 2010 - Hockenberry 1
Metabolic functions of
oncogenes
Myc
Bcl-XL
MMIG - 9 June 2010 - Hockenberry 2
Hsp90 and proteasome - dependent
degradation of mitochondrial proteins
(an ERAD-like pathway)
Mitochondrial graveyard
MMIG - 9 June 2010 - Hockenberry 3
or nursery
PCr
MbO2
ATP
HbO2
NMR
OPTICS
OPTICS
PCr
Pi
ATP
1.0
oxy
0.5
deoxy
0.0
500
ppm
MMIG - 9 June 2010 - Marcinek 1
600
700
nm
800
Less Efficient Mitochondria in Aged Muscle
H+
O2 H O
2
O2 consumption
10
ATP
*
30
*
20
4
2
ATP production
8
6
H+
H+
P/O
1
10
2
0
0
7 month
30 month
7 month
Marcinek et al., 2005, J Physiol, 569
30 month
0
7-month
MMIG - 9 June 2010 - Marcinek 2
30-month
Control of Mitochondria by Oxidative
Stress
2 wk
PQ
7 mo
SOD1-/-
Implications:
mTOR signaling, muscle atrophy, aging
apoptosis, mitochondrial biogenesis
MMIG - 9 June 2010 - Marcinek 3
HIV-related Fatigue and Mitochondrial Dysfunction
related to Antiretroviral Treatments
MMIG - 9 June 2010 - Voss 1
Projects
Proteomic Biomarker Discovery in Plasma of HIV
Patients (Dave Goodlett, School of Pharmacy)
2. Mitochondrial Function in skeletal muscle of men
with prostate cancer-related fatigue
3. Mitochondrial Dysfunction and NRTI-treatment
effects in skeletal muscle of aging mice (Dave
Marcinek, Peter Rabinovitch, School of Medicine)
4. Gene expression changes in skeletal muscle of aging
mice receiving antiretroviral treatment for 8 weeks
(Molecular Lab in the SON)
1.
MMIG - 9 June 2010 - Voss 2
Preliminary Results
 Inner mitochondrial membrane proteins in the plasma
of patients with HV-related fatigue
 Steroid hormone pathway is impacted by HIV and
ART treatment
 PGC1α and Thymidine kinase 2 are affected by AZT
treatment
 Significant changes in ATP production with AZT
treatment
MMIG - 9 June 2010 - Voss 3
CLINICAL TRIALS - FDA

THYROID HORMONE SUPPLEMENTATION IN
INFANTS UNDERGOING
CARDIOPULMONARY BYPASS (FDA)

MULTI-SITE

RECRUITED 200 PATIENTS…


ONE OF THE LARGEST CLINICAL TRIALS
EVER PERFORMED IN PEDIATRIC
CARDIOLOGY/CARDIAC SURGERY
T3 SUPPLEMENTATION IMPROVED
CLINICAL ENDPOINTS AND CARDIAC
FUNCTION IN INFANTS
MMIG - 9 June 2010 - Portman 1
MECHANISMS FOR CLINICAL RESPONSE:
ANIMAL MODELS OF PEDIATRIC
MECHANICAL-CIRCULATORY SUPPORT
Leucine
Alanine
Pyruvate
Fatty Acids
Acetyl-CoA
FA- odd carbon
Oxaloacetate
Proprionyl-CoA
Aspartate
Citrate
Malate
Isoleucine, Valine
Cis-aconitate
Phenylalanine
Isocitrate
Fumarate
CARDIAC
FUNCTION
-ketoglutarate
Succinate
T3
NiOx
glutamate
Succinyl-CoA
General scheme of
integration between
metabolism and protein
synthesis studied in
experimental plan
MMIG - 9 June 2010 - Portman 2
Glutamine
mTOR
Amino acids
mTOR-Active
Protein
AGING


Dominant negative thyroid hormone receptor alters
substrate metabolism with reduction in FFA flux
Aging in mice results in reduced FFA flux with
decreased systolic function
T3 reduced in aging mouse model
TOTAL T3 (ng/dl)
1

0.9
0.8
*
0.7
0.6
Hypothesis -
T3 treatment
modifies substrate flux and
restores cardiac function
MMIG - 9 June 2010 - Portman 3
0.5
0.4
0.3
0.2
0.1
0
YOUNG
OLD
Tian Lab studies the Heart
• Cardiac metabolism and Energetics:
• Modulating cardiac energy metabolism to improve cardiac
function during stresses – Transgenic mice and NMR
• Mitochondrial Biogenesis and Function:
• Regulation of mitochondrial biogenesis in heart failure -mtDNA replication
• Mechanistic link between mito dysfunction and the
development of pathological hypertrophy
• Metabolic Signaling:
• AMPK-activated Protein Kinase (AMPK): Signaling
cascade and isoform-specific roles of AMPK in the heart
• Metabolites regulate cardiac biology: integrating
transcriptomics with metabolomics
MMIG - 9 June 2010 - Tian 1
Tian
Functional and Metabolic Phenotyping
Transg
enic
Mice
TAC or
MI
NMR imaging +
localized
spectroscopy
Langendorff
In vivo
Perfusions
Cardiac
functio
31P NMR
n
Spectroscopy
13C
NMR
Spectroscop
y
mixed
substrates
5.5mM 1,6-13C GLU
0.4mM U-13C FA
1.2mM Lactate
50μU/ml Insulin
MMIG - 9 June 2010 - Tian 2
Tian
% oxidation of
carbon substrate by
13C NMR
WT
endo
glucose
ketones
Dynamic changes of myocardial
energetics and ATP synthesis in
beating hearts by 31P NMR
P
M
i
∞
fatty acids
PC
r
PPAR-/-
g
Pi
endo
AT
P

b
glucose
lactate
M
0
ketones
fatty acids
PPAR-/-GLUT1
endo
lactate
glucose
ketones
fatty acids
ADP + Pi  ATP
lactate
Kfor =
glucose
Flux
= Kfor x [Pi]
ketones
M0 - M∞
T 1 M∞
MMIG - 9 June 2010 - Tian 3
Tian