1. Welcome to MMIG (Mitochondria and Metabolism Interest Group)

2. 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

3. 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

4. Home base: Mitochondria and Metabolism Center at SLU http://depts.washington.edu/mmcslu http://depts.washington.edu/mmcslu Program Specialist: Karen Liebert mmcslu@u.washington.edu mmcslu@u.washington.edu –Refer new MMIG members –Suggest future meeting place –Suggest speakers, meeting time and frequency

5. gas-1 mev-1 clk-1 isp-1 ctb-1 Mitochondrial Mutants MMIG - 9 June 2010 - Morgan/Sedensky 1

6. Affected complex in kids? MMIG - 9 June 2010 - Morgan/Sedensky 2

7. Respiratory Supercomplexes in C. elegans (Suthammarak et al, JBC 2008) MMIG - 9 June 2010 - Morgan/Sedensky 3

8. 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

9. MMIG - 9 June 2010 - Morrison 2

10. MMIG - 9 June 2010 - Morrison 3

11. 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 Ca 2+ and mitochondrial membrane potential;  Assessment of cardiac myocyte function: contractility & Ca 2+ 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

12. Time (s) 488 nm 405 nm 0.3  F/F 0 10 s 488 nm 405 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/F 0 MMIG - 9 June 2010 - Wang 2

13. Integrative Study of Mitochondrial Function in Health & Disease https://depts.washington.edu/mmcslu/home MMIG - 9 June 2010 - Wang 3

14. Ca IP 3 R Ca PLC IP 3 Ca  m loss Apoptosis Ca ROS NF-  Inflammation DAG PKC Bioenergetics TCA How do mitochondria produce ROS? How are mitochondrial ROS ‘decoded’? MMIG - 9 June 2010 - Hawkins 1

15. Relationship between cytosolic calcium and mitochondrial ROS ROS MMIG - 9 June 2010 - Hawkins 2

16. 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

17. Mitochondrial hub in apoptosis Smart bombs MMIG - 9 June 2010 - Hockenberry 1

18. Myc Bcl-X L Metabolic functions of oncogenes MMIG - 9 June 2010 - Hockenberry 2

19. Hsp90 and proteasome - dependent degradation of mitochondrial proteins (an ERAD-like pathway) Mitochondrial graveyard or nursery MMIG - 9 June 2010 - Hockenberry 3

20. ATP PCr MbO 2 HbO 2 PCr ATP PiPi ppm NMR OPTICS nm oxy deoxy OPTICS MMIG - 9 June 2010 - Marcinek 1

21. Less Efficient Mitochondria in Aged Muscle Marcinek et al., 2005, J Physiol, 569 ATP H+H+ O2O2 H2OH2O H+H+ H+H+ P/O 30-month7-month * O 2 consumption * ATP production MMIG - 9 June 2010 - Marcinek 2

22. Control of Mitochondria by Oxidative Stress 2 wk PQ 7 mo SOD1 -/- mTOR signaling, muscle atrophy, aging apoptosis, mitochondrial biogenesis Implications: MMIG - 9 June 2010 - Marcinek 3

23. HIV-related Fatigue and Mitochondrial Dysfunction related to Antiretroviral Treatments MMIG - 9 June 2010 - Voss 1

24. Projects 1. 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) MMIG - 9 June 2010 - Voss 2

25. 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

26. CLINICAL TRIALS - FDA l THYROID HORMONE SUPPLEMENTATION IN INFANTS UNDERGOING CARDIOPULMONARY BYPASS (FDA) l MULTI-SITE l RECRUITED 200 PATIENTS… l ONE OF THE LARGEST CLINICAL TRIALS EVER PERFORMED IN PEDIATRIC CARDIOLOGY/CARDIAC SURGERY l T3 SUPPLEMENTATION IMPROVED CLINICAL ENDPOINTS AND CARDIAC FUNCTION IN INFANTS MMIG - 9 June 2010 - Portman 1

27. MECHANISMS FOR CLINICAL RESPONSE: ANIMAL MODELS OF PEDIATRIC MECHANICAL-CIRCULATORY SUPPORT T3 NiOx MMIG - 9 June 2010 - Portman 2

28. AGING Dominant negative thyroid hormone receptor alters substrate metabolism with reduction in FFA flux l Aging in mice results in reduced FFA flux with decreased systolic function l T3 reduced in aging mouse model Hypothesis - T3 treatment modifies substrate flux and restores cardiac function MMIG - 9 June 2010 - Portman 3

29. 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

30. Functional and Metabolic Phenotyping 13 C NMR Spectroscop y Transg enic Mice TAC or MI In vivo Cardiac functio n 31 P NMR Spectroscopy mixed substrates 5.5mM 1,6- 13 C GLU 0.4mM U- 13 C FA 1.2mM Lactate 50μU/ml Insulin Langendorff Perfusions NMR imaging + localized spectroscopy Tian MMIG - 9 June 2010 - Tian 2

31. % oxidation of carbon substrate by 13 C NMR glucose lactate glucose fatty acids endo ketones lactate WT ketones Dynamic changes of myocardial energetics and ATP synthesis in beating hearts by 31 P NMR fatty acids endoglucose lactate ketones PPAR  -/- glucose fatty acids endo lactate PPAR  -/- GLUT1 ketones PiPi PiPi PC r AT P    M∞M∞ M0M0 ADP + P i  ATP K for = M0 - M∞M0 - M∞ T 1 M ∞ K for x [P i ]Flux = Tian MMIG - 9 June 2010 - Tian 3