Novel Targets to Attack Exercise Intolerance in Heart Failure - Skeletal Muscle - Volker Adams, PhD ESC, Paris 3. Aug. 211 UNIVERSITÄT LEIPZIG H E R Z Z E N T R U M Nothing to disclose
Myers et al. NEJM 22 Exercise Capacity and Mortality
Adjusted all cause mortality Exercise Capacity and Mortality 2867 men perform an exercise test (Bruce protocol or ramp test) Follow-up period 22.8 years (mean 7.8±5.1 years) 5 65 yrs (n=1 658) > 65 yrs (n=1 29) 1..8.6.69.7.4.36.36.42.2.2 < 4. 4.1-6. 6.1-8. > 8. METs < 4. 4.1-6. 6.1-8. > 8. METs Nylen et al. J Am Geriatr Soc 21
central hemodynamic pulmonal function Exercise training in healthy and diseased skeletal muscle endothelial function
Targets of Exercise Training Inflammation - Cytokines Anabolic/catabolic factors Apoptosis Oxidative stress Energy Metabolism Stem cells Capillarization Micro-RNA
Targets of Exercise Training Inflammation - Cytokines Anabolic/catabolic factors Apoptosis Oxidative stress Energy Metabolism Stem cells Capillarization Micro-RNA
Effects of Exercise Training on Circulating Cytokine Levels Randomized study, 24 patients with CHF (NYHA II/III) and 2 controls; exercise training over a period of 12 weeks. Sampling of blood at begin and after 12 weeks. TNF-a (pg/ml) IL-6 (pg/ml) 1. p<.5 1. p<.5 7.5 7.5 5. p 5. p 2.5 2.5. Beg. 12 w Beg. 12 w Control CHF. Beg. 12 w Beg. 12 w Control CHF Adamopoulos et al., J Am Coll Cardiol 22
Local IL-6 Expression [rel. units] Local TNF-a Expression [rel. units] Control Training Local IL-1ß Expression [rel. units] Effects of Exercise Training on Local Cytokine Expression 3 5 4 2 1 # 3 2 * 1 Begin 6 Months Begin 6 Months 8 6 4 2 $ * p<.5 versus Control $ p<.5 versus Begin # p<.5 for D versus Control Begin Gielen S et al., J Am Coll Cardiol 23 6 Months
Targets of Exercise Training Inflammation - Cytokines Anabolic/catabolic factors Apoptosis Oxidative stress Energy Metabolism Stem cells Capillarization Micro-RNA
Ubiquitin proteasome pathway chronic heart failure denervation immobilisation weightlessness Murf 1/ MAFbx (E3 ubiqitin ligases) proteins proteins ubiquitin ubiquitin protein complex skeletal muscle cell proteolysis
Study Design - Exercise Training and E3 Ligase Expression CHF group LAD Ligation sham OP sham group Exercise Training (3weeks, 2x per day, 3 min) Control Exercise Training (3weeks, 2x per day, 3 min) Control Analysis of Gene Expression
mrna expression [arb. Units] mrna expression [arb. Units] Exercise Training and MAFbx / Murf-1 Expression MAFbx Murf-1 3 P<.5 P<.1 4 P<.5 P<.1 3 2 p 2 p 1 1 sham-c sham-t CHF-C CHF-T sham-c sham-t CHF-C CHF-T
Murf1 /18S Correlation of Murf-1 Expression and TNF-alpha 2 r=.62 P<.1 1 5 1 15 2 TNF - a [ pg /mg Protein] Adams et al. Cardiovasc Res 27
Change in Murf-1 expression [%] Change in Murf-1 expression by exercise training - NYHA III Study D Muscle mass (%) 5 * p<.5 2 15 r=.62 25 1 Control p Training 5-15 -1-5 5 1 15 2-5 p D Murf-1 (%) -25-1 -15-5 * -2
Murf-1 / 18SrRNA Change in Murf-1 expression by exercise training Leipzig exercise intervention in CHF and ageing trail (LEICA) Murf-1 expression % change in Murf-1 expression 3 p<.5 5 p<.5 25 2 1 p Healthy control Healthy p training CHF control CHF training -25 Healthy CHF -5
Study Design and Methods WT and MuRF-KO IP injection of TNF-a (1 ng/g) IP injection of NaCl 24h inoculation time Measurement of muscle contractility (soleus, force-frequency relation) Expression analysis of MuRF1 and contractile proteins
Force [g/sq cm] Force Frequency Relation in Soleus Muscle 3 25 2 15 WT control Murf-1 -/- control Murf-1 -/- + TNF alpha WT + TNF alpha 1 5 1 Hz 1 Hz 25 Hz 5 Hz 75 Hz 1 Hz 125 Hz 15 Hz Frequency [Hz] Murf-1 -/- + TNF alpha WT + TNF alpha WT control Murf-1 -/- control Adams et al., J Mol Biol 28
Pathophysiological Model for the TNF-a Induced Reduction in Muscle Contractility Dexa NF-kB TNNT-3 TNF - a p38 MAPK Murf - 1 Translation Elongation Muscle contractility ROS Adams et al., J Mol Biol 28
Myostatin knock out animal Belgian blue Myostatin negative regulator of muscle mass
Myostatin protein expression (arb.units) Exercise Training and Myostatin Expression LAD Ligation sham OP 3 p<.5 p<.5 CHF group sham group 2 Exercise Training (4weeks, 2x/day, 3 min) Control 1 Analysis of myostatin protein expression sham CHF CHF sedentary CHF training Lenk et al., Eur J Heart Fail 29
IGF-I mrna Expression [ar. Units] IGF-1 and Skeletal Muscle Mass IGF-1 transgenic animals (IGF-1 under the control of skeletal muscle promotor) Local IGF-I Expression vs. Muscle Cross-Sectional Area 15 r=.75 p=.1 1 5 4 5 6 7 8 Muscle CSA [cm 2 ] Musaro et al.; Nature 21 Hambrecht et al.; J Am Coll Cardiol 22
[% positive tissue area] Effects of Exercise Training on Local IGF-I Expression in CHF.3 IGF-I mrna Expression * p <.1 vs. Control * IGF-I Protein Content * p <.5 vs. Control 1.5 *.2 1..1.5 Begin 6 months Begin 6 months Training Control Hambrecht et al.; Eur J Cardiovasc Prev Rehab 25
Targets of Exercise Training Inflammation - Cytokines Anabolic/catabolic factors Apoptosis Oxidative stress Energy Metabolism Stem cells Capillarization Micro-RNA
Production and Detoxification of Reactive Oxygen Species Production Detoxification NAD(P)H oxidase xanthine oxidase myeloperoxidase uncoupled NOS Enzymes of the respiratory chain SOD O 2 - H 2 O 2 GSSG GSH GPX 2 H 2 O Fe 2+ Fe 3+ OH Catalase H 2 O + O 2
enzyme activity (U/mg) Lipid peroxides (µmol/mg) enzyme activity (U/mg) enzyme activity (U/mg) Effects of Exercise Training on Oxidative Stress - Radical Scavenger Enzymes - SOD 1. GPX 1 7.5 5. 2.5 75 5 25 * * Catalase. 4 HS B 6Mo CHF Training B 6Mo CHF Control p 6 HS B 6Mo CHF Training B 6Mo CHF Control 3 5 * * 2 1 * * 4 3 2 1 HS B 6Mo B 6Mo CHF Training CHF Control HS B 6Mo B 6Mo CHF Training CHF Control Linke et al., Circulation 25 * p<.5 vs. HS p<.5 vs. CHF Training B
D ROS generation in leucocytes (%) Effects of Exercise Training on ROS Leipzig exercise intervention in CHF and ageing trail (LEICA) DHE 5 p<.1 25 ROS DHE DHE ox Healthy control Healthy training CHF control CHF training -25 DHE: Dihydroxyethidium, is able to pass the cell membrane, non-fluorescent DHE ox : Dihydroxyethidium oxidized form, is no longer able able to pass the cell membrane, fluorescent FACS -5
Targets of Exercise Training Inflammation - Cytokines Anabolic/catabolic factors Apoptosis Oxidative stress Energy Metabolism Stem cells Capillarization Micro-RNA
Capillaries / muscle fiber Capillary Density in Skeletal muscle of CHF Rabbit model of CHF > LAD ligation Patients with CHF 1.5 p<.1 1.5 p<.1 1..5 Capillaries / mm 2 1..5 Healthy CHF Healthy CHF Nusz et al., Am J Physiol 23 Williams et al., J Card Fail 24
Capaillaries / muscle fibre Impact of Exercise Training on Capillary Density in the Skeletal Muscle in Patients With CHF NYHA III Study D Capillary Density (%) 25 2 p<.5 2 p<.5 vwf staining 15 1 5 1 Training Control -5-1 Beg. 12 w. Beg. 12 w. Training Control -15 Erbs et al., Circ Heart Fail 21
Targets of Exercise Training Inflammation - Cytokines Anabolic/catabolic factors Apoptosis Oxidative stress Energy Metabolism Stem cells Capillarization Micro-RNA
Definition of micro-rna mirnas are small (~ 2-3 nucleotides) non-coding RNAs which are highly conserved from plants to mammals. Their known functions are to inhibit protein translation or to enhance mrna degradation. Mammalian genomes are predicted to encode 2 to 5 unique mirnas.
Micro-RNAs in the Skeletal Muscle and Exercise Training Endurance Resistance (single bout) single bout Training mir-1 mir-133a mir-26 mir-23a mir-1 mir-181 mir-17 mir-696 mir-21 mir-79 mir-72 Güller and Russell., J Physiol 21
mir-696 expression PGC-1a expression PGC-1a expression (arb Units) Targets for Specific mirna 3 r=.64 2 sedentary group training group (4wk) 1 mir-696 PGC-1a 12 p<.5 15 p<.5 1 2 3 mir-696 expression (arb Units) 8 1 Training Aerobic metabolism 4 5 mir-696 Mitochondrial content Fatty acid oxidation sedentary training sedentary training PGC-1a Fiber type shift
Targets of Exercise Training Inflammation - Cytokines Anabolic/catabolic factors Apoptosis Oxidative stress Energy Metabolism Stem cells Capillarization Micro-RNA