welcome to mmig (mitochondria and metabolism interest group)

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Welcome to MMIG (Mitochondria and Metabolism Interest Group)

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Page 1: Welcome to MMIG (Mitochondria and Metabolism Interest Group)

Welcome to MMIG

(Mitochondria and Metabolism Interest Group)

Page 2: 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

Page 3: Welcome to MMIG (Mitochondria and Metabolism Interest Group)

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

Page 4: Welcome to MMIG (Mitochondria and Metabolism Interest Group)

• 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

Page 5: Welcome to MMIG (Mitochondria and Metabolism Interest Group)

gas-1

mev-1

clk-1

isp-1ctb-1

Mitochondrial Mutants

MMIG - 9 June 2010 - Morgan/Sedensky 1

Page 6: Welcome to MMIG (Mitochondria and Metabolism Interest Group)

Affected complex in kids?

EC50s for BIS=60

0

1

2

3

4

5

Mitochondrial Defects

% s

evo

flu

ran

e

complex I

non complex I

normal

MMIG - 9 June 2010 - Morgan/Sedensky 2

Page 7: Welcome to MMIG (Mitochondria and Metabolism Interest Group)

Respiratory Supercomplexes in C. elegans

Respiratory Supercomplexes in C. elegans

(Suthammarak et al, JBC 2008)MMIG - 9 June 2010 - Morgan/Sedensky 3

Page 8: Welcome to MMIG (Mitochondria and Metabolism Interest Group)

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?

The complexities of regulating mitochondria in neurons

MMIG - 9 June 2010 - Morrison 1

Page 9: Welcome to MMIG (Mitochondria and Metabolism Interest Group)

MMIG - 9 June 2010 - Morrison 2

Page 10: Welcome to MMIG (Mitochondria and Metabolism Interest Group)

MMIG - 9 June 2010 - Morrison 3

Page 11: Welcome to MMIG (Mitochondria and Metabolism Interest Group)

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

Page 12: Welcome to MMIG (Mitochondria and Metabolism Interest Group)

Time (s)

488 nm

405 nm

0.3

F/F

0

10 s488 nm405 nm

10 m

Single Mitochondrial Superoxide Flash in Adult Cardiac Myocyte

(Wang et al, 2008, Cell 134:279-290)

TMRM

488 nm

405 nm

20 s

0.5 F

/F0

MMIG - 9 June 2010 - Wang 2

Page 13: Welcome to MMIG (Mitochondria and Metabolism Interest Group)

Integrative Study of Mitochondrial Function in Health & Disease

https://depts.washington.edu/mmcslu/homeMMIG - 9 June 2010 - Wang 3

Page 14: Welcome to MMIG (Mitochondria and Metabolism Interest Group)

Ca

Ca

Ca

CaCa

Ca

Ca Ca

Ca

IP3R

Ca

Ca

Ca

Ca

Ca

CaCa

Ca

Ca

Ca

Ca

Ca

Ca

Ca

Ca

Ca

Ca

Ca

PLC

IP3

Ca

Ca

Ca Ca

Ca Ca

m loss

Apoptosis

Ca

CaCa

Ca

Ca

Ca

CaCa

ROS

NF-

Inflammation

DAGPKC

Bioenergetics

TCA

How do mitochondria produce ROS?How are mitochondrial ROS ‘decoded’?

MMIG - 9 June 2010 - Hawkins 1

Page 15: Welcome to MMIG (Mitochondria and Metabolism Interest Group)

Relationship between cytosolic calcium and mitochondrial ROS ROS

MMIG - 9 June 2010 - Hawkins 2

Page 16: Welcome to MMIG (Mitochondria and Metabolism Interest Group)

Laboratory Interests• How do disparate mitochondrial calcium uptake patterns translate to cellular

function?

• How do calcium-linked oxidants influence mitochondrial function?

• What are the cellular targets of mitochondrial reactive oxygen species?

Microvascular EC

Macrovascular EC

MMIG - 9 June 2010 - Hawkins 3

Page 17: Welcome to MMIG (Mitochondria and Metabolism Interest Group)

Mitochondrial hub in apoptosis

Smart bombs

MMIG - 9 June 2010 - Hockenberry 1

Page 18: Welcome to MMIG (Mitochondria and Metabolism Interest Group)

Myc

Bcl-XL

Metabolic functions of oncogenes

MMIG - 9 June 2010 - Hockenberry 2

Page 19: Welcome to MMIG (Mitochondria and Metabolism Interest Group)

Hsp90 and proteasome - dependentdegradation of mitochondrial proteins(an ERAD-like pathway)

Mitochondrial graveyard or nurseryMMIG - 9 June 2010 - Hockenberry

3

Page 20: Welcome to MMIG (Mitochondria and Metabolism Interest Group)

ATP

PCr

MbO2

HbO2

PCr

ATP

Pi

ppm

NMROPTICS

500 600 700 8000.0

0.5

1.0

nm

oxy

deoxy

OPTICS

MMIG - 9 June 2010 - Marcinek 1

Page 21: Welcome to MMIG (Mitochondria and Metabolism Interest Group)

Less Efficient Mitochondria in Aged Muscle

Marcinek et al., 2005, J Physiol, 569Marcinek et al., 2005, J Physiol, 569

ATP

H+

O2 H2O

H+ H+

P/O

30-month7-month0

1

2

*

0

2

4

6

8

10

7 month 30 month

O2 consumption

0

10

20

30

7 month 30 month

*

ATP production

MMIG - 9 June 2010 - Marcinek 2

Page 22: Welcome to MMIG (Mitochondria and Metabolism Interest Group)

Control of Mitochondria by Oxidative Stress

2 wkPQ

7 moSOD1-/-

mTOR signaling, muscle atrophy, agingapoptosis, mitochondrial biogenesis

Implications:

MMIG - 9 June 2010 - Marcinek 3

Page 23: Welcome to MMIG (Mitochondria and Metabolism Interest Group)

HIV-related Fatigue and Mitochondrial Dysfunction related to Antiretroviral

Treatments

MMIG - 9 June 2010 - Voss 1

Page 24: Welcome to MMIG (Mitochondria and Metabolism Interest Group)

Projects1. 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 fatigue3. 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)

MMIG - 9 June 2010 - Voss 2

Page 25: Welcome to MMIG (Mitochondria and Metabolism Interest Group)

Preliminary ResultsInner mitochondrial membrane proteins in

the plasma of patients with HV-related fatigueSteroid hormone pathway is impacted by HIV

and ART treatmentPGC1α and Thymidine kinase 2 are affected

by AZT treatmentSignificant changes in ATP production with

AZT treatment

MMIG - 9 June 2010 - Voss 3

Page 26: Welcome to MMIG (Mitochondria and Metabolism Interest Group)

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

Page 27: Welcome to MMIG (Mitochondria and Metabolism Interest Group)

MECHANISMS FOR CLINICAL RESPONSE: ANIMAL MODELS OF PEDIATRIC MECHANICAL-CIRCULATORY SUPPORT

Oxaloacetate

Citrate

Cis-aconitate

Isocitrate

-ketoglutarate

Succinyl-CoA

Succinate

Fumarate

Malate

glutamate

Glutamine

Acetyl-CoAPyruvateFatty Acids

Aspartate

Proprionyl-CoA

mTOR mTOR-Active

Phenylalanine

Isoleucine, Valine

ProteinAmino acids

CARDIAC FUNCTION

Leucine

FA- odd carbon

General scheme of integration between metabolism and protein synthesis studied in experimental plan

Alanine

T3NiOx

MMIG - 9 June 2010 - Portman 2

Page 28: Welcome to MMIG (Mitochondria and Metabolism Interest Group)

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 modelTOTAL T3 (ng/dl)

0

0.1

0.2

0.3

0.4

0.5

0.6

0.7

0.8

0.9

1

YOUNG OLD

*

TOTAL T3 (ng/dl)

0

0.1

0.2

0.3

0.4

0.5

0.6

0.7

0.8

0.9

1

YOUNG OLD

*

Hypothesis - T3 treatment modifies substrate flux and restores cardiac function

MMIG - 9 June 2010 - Portman 3

Page 29: Welcome to MMIG (Mitochondria and Metabolism Interest Group)

• 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

Tian Lab studies the Heart

Tian MMIG - 9 June 2010 - Tian 1

Page 30: Welcome to MMIG (Mitochondria and Metabolism Interest Group)

Functional and Metabolic Phenotyping

13C NMR Spectroscop

y

Transgenic Mice

TAC or MI

In vivo Cardiac functio

n31P NMR

Spectroscopy

mixed substrates

5.5mM 1,6-13C GLU 0.4mM U-13C FA 1.2mM Lactate 50μU/ml Insulin

Langendorff Perfusions

NMR imaging + localized

spectroscopy

NMR imaging + localized

spectroscopy

Tian MMIG - 9 June 2010 - Tian 2

Page 31: Welcome to MMIG (Mitochondria and Metabolism Interest Group)

% oxidation of carbon substrate by 13C NMR

glucose

lactate

glucose

fatty acids

endo

ketones

lactate

WT

ketones

Dynamic changes of myocardial energetics and ATP synthesis in beating hearts by 31P NMR

fatty acids

endo

glucoselactate

ketones

PPAR-/-

glucose

fatty acids

endo

lactate

PPAR-/-GLUT1

ketones

P

i

Pi

PCr AT

P

M

M

0

ADP + Pi ATP

Kfor =M0 - M∞

T1 M∞

Kfor x [Pi]Flux =

Tian MMIG - 9 June 2010 - Tian 3