Abstract 232: Bendavia, a Novel Mitochondria Targeting Peptide, Improves Mitochondrial Function in Circulating Blood Monocytes From Dogs With Chronic Heart Failure

2015 ◽  
Vol 117 (suppl_1) ◽  
Author(s):  
Hani N Sabbah ◽  
Vinita Sing-Gupta ◽  
Ramesh C Gupta
Keyword(s):  
Heart Failure ◽  
Atp Synthesis ◽  
Proton Leak ◽  
Blood Monocytes ◽  
Consumption Rates ◽  
Presence And Absence ◽  
Targeting Peptide ◽  
Dose Dependent Decrease ◽  
Measure Oxygen Consumption ◽  
Dose Dependent

Background: Mitochondrial (MITO) function is abnormal in heart failure (HF) as evidenced by reduced MITO respiration and rate of ATP synthesis. We showed that MITO dysfunction can be normalized in HF dogs after therapy with Bendavia (BEN), a novel MITO targeting peptide. This study tested the hypothesis that BEN will also reverse abnormalities of MITO function present in blood monocytes (MCs) of HF dogs. Methods: Blood samples obtained from 6 normal (NL) dogs and 6 dogs with coronary microembolizations-induced HF (LV ejection fraction ~30%) were used to isolate MCs by sequential Ficoll and Percoll density gradients. An XFe/XF96 analyzer (Seahorse Bioscience) was used to measure oxygen consumption rates (OCR) in MCs in the presence and absence of 1 μM oligomycin, 0.5 μM FCCP, or 1 μM each rotenone and antimycin. MITO proton leak, maximal respiration (MAXresp) and spare respiratory capacity (SRC) were measure in the presence and absence of 0.1, 1.0 and 10 μM concentrations of BEN and results expressed in pmols OCR/min/μg protein. Results: Proton leak, MAXresp and SRC were abnormal in MCs of HF dogs compared to NL. Incubation with BEN had no effect on measures of MCs MITO function of NL dogs but nearly normalized MITO function of MCs of HF dogs as evidenced by a dose-dependent increase MAXresp and SRC and dose-dependent decrease in proton leak (Table). Conclusions: MITO function is abnormal in blood MCs of HF dogs. BEN had no effect on MITO function of MCS from NL dogs but normalized MITO function in MCs from HF dogs. These findings support the use of circulating blood MCs as means of assessing MITO dysfunction in HF and as a marker of potential benefits derived from treatment with MITO targeted therapies.

Abstract 90: Bendavia (MTP-131), a Novel Mitochondria-Targeting Peptide, Improves ADP-Stimulated Mitochondrial Respiration in Cardiomyocytes Isolated From Dogs with Chronic Heart failure

2014 ◽  
Vol 115 (suppl_1) ◽  
Author(s):  
Hani Sabbah ◽  
Ramesh C Gupta ◽  
Sharad Rastogi ◽  
Paula Mohyi ◽  
Kristina Szekely
Keyword(s):  
Heart Failure ◽  
Mitochondrial Respiration ◽  
Systolic Function ◽  
Atp Synthesis ◽  
Vehicle Control ◽  
Electrode System ◽  
Lv Function ◽  
Targeting Peptide ◽  
Lv Systolic Function ◽  
Mitochondria Targeting

Background: Mitochondria (MITO) of failed human hearts and hearts of dogs with experimental heart failure (HF) manifest structural and functional abnormalities characterized by hyperplasia and reduced organelle size and reduced respiration. These abnormalities lead to reduced ATP synthesis that adversely impacts LV function. We previously showed that chronic therapy (3 months) with Bendavia (MTP-131), a novel mitochondria-targeting peptide, improves LV systolic function in dogs with heart failure (HF), reverses MITO abnormalities and normalizes mitochondria ATP synthesis in myocardium from Bendavia-treated HF dogs. In the present study we examined the direct effects of Bendavia on mitochondria ADP-stimulated state 3 respiration in freshly isolated cardiomyocytes from dogs with advanced chronic HF. Methods: Cardiomyocytes were isolated from LV free wall of 3 untreated dogs with HF produced by intracoronary microembolizations (LV ejection fraction <30%). A standard collagenase-based enzymatic process was used for isolation that yielded ~70% viable rod-shaped cardiomyocytes that excluded trypan blue. Equal aliquotes of cardiomyocytes were incubated in 0, 0.01, 0.10, 1.0 and 10 μM concentration of Bendavia for one hour at 37°C. At the end of incubation, ADP-stimulated state-3 respiration was measured using a Clark electrode system and quatified in nAtom Oxygen/min/mg protein. Results: State-3 respiration in the absence of Bendavia (Vehicle-Control) was 248±9 nAtom Oxygen/min/mg protein. Compared to vehicle-control, incubation of failing cardiomyocytes with Bendavia significantly increased state-3 respiration to 303±33 at 0.01 μM, p<0.05; 405±39 at 0.10 μM, p<0.05; 371±28 at 1.0 μM, p<0.05; and 346±29 at 10.0 μM, p<0.05. Conclusions: Results of this study indicate that the effects of Bendavia on mitochondrial respiration in cardiomyocytes is direct and not a consequence of improved global LV structure or function. Furthermore, the results indicate that the improvement in mitochondrial respiration after treatment with Bendavia can occur early after initiation of therapy (within one hour) and is dose-dependent up to concentrations of 0.10 μM.

Dose-dependent decrease of platelet activation and tissue factor by omega-3 polyunsaturated fatty acids in patients with advanced chronic heart failure

2011 ◽  
Vol 106 (09) ◽  
pp. 457-465 ◽  
Author(s):  
Rudolf Berger ◽  
Alexandra Hammer ◽  
Raisa Hutuleac ◽  
Renate Koppensteiner ◽  
Christoph Kopp ◽  
...  
Keyword(s):  
Heart Failure ◽  
Fatty Acids ◽  
Chronic Heart Failure ◽  
Polyunsaturated Fatty Acids ◽  
Platelet Activation ◽  
Interleukin 6 ◽  
Omega 3 ◽  
Chemotactic Protein ◽  
Dose Dependent Decrease ◽  
Dose Dependent

SummaryChronic heart failure (CHF) is characterised by activation of neuroendocrine and inflammatory pathways, and both are linked to a prothrombotic state. Treatment with omega-3 polyunsaturated fatty acids (n3-PUFA) showed significant benefits including mortality reduction in CHF, but exact mechanisms of action are still unclear. We investigated the effects of n3-PUFA on markers of platelet activation and thrombogenesis in patients with severe CHF. Thirty-six patients with non-ischaemic CHF (LVEF<35%, NYHA class>2) under optimised therapy were randomised to supplementation with 1g/day or 4g/day n3-PUFA, or placebo for 12 weeks. Using whole-blood flow cytometry, monocyteplatelet aggregates characterised by CD14+/CD42b+ co-expression and monocytic tissue factor (TF) were determined. Plasma levels of P-selectin, sCD40L, fibrinogen, prothrombin fragment F1.2, TF and proinflammatory markers (high sensitive[hs] interleukin-6, hsCRP, hsTNFalpha, monocyte chemotactic protein-1) were measured by immunoassay. Supplementation with 1g/day and 4g/day n3-PUFA but not placebo significantly reduced monocyte-platelet aggregates in a dose-dependent manner (p for trend=0.02 across the groups). A dose of 4g/day but not 1g/day n3-PUFA significantly decreased P-selectin (p=0.03). Plasma TF decreased dose-dependently upon n3-PUFA supplementation (p for trend=0.02), paralleled by a significant decrease of TF+-monocytes (p for trend=0.01). The amount of 4g/day n3-PUFA exhibited modest anti-inflammatory effects with a significant reduction of hs interleukin-6 (p<0.01) and a trend-wise reduction of hsTNF-alpha (p=0.09). No changes were seen for sCD40L, fibrinogen, hsCRP and monocyte chemotactic protein-1, while F1.2 was decreased by 4g/day n3-PUFA (P=0.03). In patients with severe non-ischaemic CHF, treatment with n3-PUFA leads to a dose-dependent decrease of platelet activation and TF. Higher dosage exhibits also anti-inflammatory effects.* ClinicalTrials.gov registration number: NCT00149409

scholarly journals Blunted nitric oxide-mediated inhibition of renal nerve discharge within PVN of rats with heart failure

2001 ◽  
Vol 281 (3) ◽  
pp. H995-H1004 ◽  
Author(s):  
Kun Zhang ◽  
Yi-Fan Li ◽  
Kaushik P. Patel
Keyword(s):  
Nitric Oxide ◽  
Heart Failure ◽  
Blood Pressure ◽  
No Synthase ◽  
Coronary Artery Ligation ◽  
Artery Ligation ◽  
Renal Nerve ◽  
Dose Dependent Decrease ◽  
Dose Dependent ◽  
Nerve Discharge

We have demonstrated a decreased neuronal nitric oxide (NO) synthase (nNOS) message in the hypothalamus of rats with heart failure (HF). Subsequently, we have demonstrated that NADPH diaphorase (a commonly used marker for nNOS activity) positive neurons are decreased in paraventricular nucleus (PVN) of rats with coronary artery ligation model of HF. The goal of the present study was to examine the influence of endogenous NO within the PVN on renal sympathetic nerve discharge (RSND) during HF. In α-chloralose- and urethane-anesthetized rats, an inhibitor of NO synthase, N G-monomethyl-l-arginine (l-NMMA) microinjected into the PVN (50, 100, and 200 pmol in 50–200 nl) produced a dose-dependent increase in RSND, blood pressure, and heart rate in control and HF rats. These responses were attenuated in rats with HF compared with control rats. On the other hand, the NO agonist, sodium nitroprusside, microinjected in PVN produced a dose-dependent decrease in RSND and blood pressure in control and HF rats. These responses were less in rats with HF compared with control rats. These data suggest that the endogenous NO-mediated effect within the PVN of HF rats is less potent in suppressing RSND compared with control rats. These data support the conclusion that the NO system within the PVN involved in controlling autonomic outflow is altered during HF and may contribute to the elevated levels of renal sympathoexcitation commonly observed in HF.

Abstract 770: Coordinated Upregulation of Cardiomyocyte Urotensin II (U-II) and its Receptor (UT) Exhibits Enhanced Inhibition of L-Type Calcium Current in Heart Failure

Circulation ◽  
2007 ◽  
Vol 116 (suppl_16) ◽  
Author(s):  
Heng-Jie Cheng ◽  
Qing-Hua Han ◽  
Peng Zhou ◽  
Atsushi Morimoto ◽  
Satoshi Masutani ◽  
...  
Keyword(s):  
Heart Failure ◽  
Cell Voltage ◽  
Urotensin Ii ◽  
Adult Rats ◽  
Rat Cardiomyocytes ◽  
Protein Levels ◽  
Cardiac Depression ◽  
Dose Dependent Decrease ◽  
Induced Changes ◽  
Dose Dependent

Background . Heart failure has been linked to urotensin II (U-II), the most potent mammalian vasoconstrictor. However, the alteration and functional significance of U-II in HF are uncertain. Methods . We compared LV myocyte U-II and its receptor (UT) protein levels and I Ca,L response to U-II stimulation in 10 normal (N) adult rats and 10 age-matched rats with isoproterenol (ISO)-induced HF (4 months after 340 mg/kg, sq, for 2 days). I Ca,L was measured using whole-cell voltage clamp technique. In 3 subgroups, U-II-mediated actions were determined after pretreatment of myocytes with U-II receptor antagonist, Urantide (10 −5 M); pertussis toxin (PTX, 2μg/ml, 36°C, 6h); and dibutyryl-cAMP (Db-cAMP, 10 −4 M). Results . Compared with normal myocytes, in HF myocytes, both U-II (93%, 0.081 vs 0.042) and UT (52%, 0.038 vs 0.025) protein levels were significantly increased. Importantly, these changes were associated with enhanced U-II -mediated negative modulation on L Ca,L . Superfusion of U-II (10 −8 -10 −5 M) caused a dose-dependent decrease in I Ca,L in both normal and HF myocytes with maximal inhibition at 10 −5 M. In normal myocytes, U-II (10 −5 M) decreased I Ca,L by 18% (5.0±0.29 vs 6.1±0.5 pA/pF, p<0.01). In HF myocytes, the baseline I Ca,L was significantly lower (3.3±0.1 vs 5.9±0.3 pA/pF, p<0.01) and was further reduced by 27% with U-II 10 −5 M (2.4±0.1 vs 3.3±0.1 pA/pF), that was significantly greater than U-II-induced changes in normal myocytes. U-II-induced reductions in I Ca,L were significantly attenuated by U-II-antagonist. In both groups of rats, U-II-induced decreases of I Ca,L were prevented with pretreatment myocytes with PTX (normal: 5.6±0.5 vs 5.5±0.4; HF: 3.4±0.4 vs 3.5±0.2 pA/pF) or Db-cAMP (normal: 5.6±0.5 vs 5.6±0.7; HF: 3.7±0.3 vs 3.8±0.4 pA/pF). Conclusions . In HF rat cardiomyocytes, U-II and UT-coupled, G i -mediated signaling pathway is upregulated, which amplifies U-II stimulation-induced inhibition of I Ca,L . This may lead to exacerbation of the dysfunctional [Ca 2+ ] i homeostasis and enhance cardiac depression of the failing myocardium.

Abstract 090: Long-term Therapy with a Bendavia (MTP-131), a Novel Mitochondria-Targeting Peptide, Normalizes Cardiolipin Biosynthesis and Remodeling in Left Ventricular Myocardium of Dogs with Advanced Heart Failure

2013 ◽  
Vol 113 (suppl_1) ◽  
Author(s):  
Hani N Sabbah ◽  
Quan He ◽  
Ramesh C Gupta ◽  
Sharad Rastogi ◽  
Mengjun Wang ◽  
...  
Keyword(s):  
Heart Failure ◽  
Atp Synthesis ◽  
Left Ventricular ◽  
Advanced Heart Failure ◽  
Term Therapy ◽  
Complex Iv ◽  
Targeting Peptide ◽  
Long Term Therapy ◽  
Mitochondria Targeting

Background: Cardiolipin (CL) is a signature phospholipid of the mitochondrial inner membrane and responsible for modulation of activities of various enzymes essential for oxidative phosphorylation. Cardiac CL is biosynthesized in a series of steps from phosphatidic acid and remodeled into a form which contains four 18:2 fatty acid chains, tetralinoleol CL [(18:2) 4 CL]. Long-term therapy with Bendavia (MTP-131), a novel mitochondria-targeting peptide, was previously shown to improve left ventricular (LV) function, increase ATP synthesis, and increase the activity and protein levels of mitochondria cytochrome c oxidase (complex IV) in LV myocardium of dogs with advanced heart failure (HF). This study examined the effects of Bendavia on total CL and (18:2) 4 CL in LV myocardium of dogs with HF. Methods: LV tissue was obtained from 12 dogs with microembolization-induced HF randomized to 3 months therapy with subcutaneous injections of Bendavia (0.5 mg/kg once daily, n=7) or saline (Control, n=7). LV tissue from 7 normal dogs was used for comparison. Total CL and (18:2) 4 CL were measured using electrospray ionization mass spectroscopy and quantified in nmol/mg of non-collagen protein (NCP). Results: Total CL was significantly decreased in LV myocardium of Control dogs compared to normal dogs (19.1±1.1 vs. 26.7±1.4 nmol/mg, p<0.05) as was (18:2) 4 CL (14.2±0.9 vs. 20.5±1.2 nmol/mg, p<0.05). Long-term therapy with Bendavia significantly increased both total CL (23.6±1.1 nmol/mg) and (18:2) 4 CL (17.8±1.0 nmol/mg) to near normal levels in LV myocardium of treated HF dogs compared to untreated HF Controls (p<0.05). Conclusions: Total CL and (18:2) 4 CL are decreased in LV myocardium of dogs with HF. Treatment with Bendavia normalizes total CL and (18:2) 4 CL. These findings are consistent with observed normalization of mitochondrial complex IV activity, normalization of rate of ATP synthesis and improvement of global LV performance in dogs with HF after long-term therapy with Bendavia.

Abstract 231: Bendavia, a Novel Mitochondria-Targeting Peptide, Improves Contraction and Relaxation of Failing Cardiomyocytes Isolated From Dogs With Chronic Heart Failure

2015 ◽  
Vol 117 (suppl_1) ◽  
Author(s):  
Hani N Sabbah ◽  
Ramesh C Gupta
Keyword(s):  
Heart Failure ◽  
Atp Synthesis ◽  
Electrical Field Stimulation ◽  
Lv Function ◽  
Shortening Velocity ◽  
Cell Contractility ◽  
Targeting Peptide ◽  
Reduced Rate ◽  
Systolic And Diastolic Function ◽  
Mitochondria Targeting

Background: Mitochondria (MITO) of humans and dogs with heart failure (HF) manifest functional abnormalities characterized by poor respiration, reduced rate of ATP synthesis and increased production of reactive oxygen species (ROS) that adversely impact LV systolic and diastolic function. We previously showed that chronic therapy with Bendavia (BEN, MTP-131), a novel mitochondria-targeting peptide, improves global LV function in dogs with HF without affecting heart rate or blood pressure. This improvement was associated with a reversal of MITO abnormalities and normalization of rate of ATP synthesis. This study tested the hypothesis that the improvement in global LV function seen in dogs with HF during treatment with BEN results primarily from enhanced function of constituent LV cardiomyocytes. Methods: Cardiomyocytes were isolated from the LV free wall of 8 untreated dogs with coronary microembolization-induced HF (LV ejection fraction <30%). A collagenase-based enzymatic process was used for the isolation and yielded ~70% viable rod-shaped cardiomyocytes that excluded trypan blue. Extent of cardiomyocytes shortening, shortening velocity and lengthening velocity were assessed during 1.0 Hz electrical field stimulation delivered via a MyoPacer (ION Optix). Measurements were made at baseline and were repeated after one hour of gradual exposure of the same cardiomyocytes to BEN at a concentration of 0.1 μM. Results: At baseline, the extent of cardiomyocyte shortening was 3.7 ± 0.8 μm, shortening velocity was 62.8 ± 16.9 μm /sec and lengthening velocity was -53.8 ± 16.5 μm/sec. Exposure of cardiomyocyte to BEN significantly increased the extent of cardiomyocyte shortening to 5.4 ± 1.1 μm (p<0.012), shortening velocity to 94.5 ± 21.9 μm/sec (p<0.020) and lengthening velocity to -96.8 ± 21.1 μm/sec (p<0.016) compared to baseline. Conclusions: Exposure of failing isolated cardiomyocytes to BEN elicits improvements in the rate of cardiomyocyte shortening and re-lengthening indicative of improved cell contractility and relaxation. This improvement is likely mediated by increased availability of ATP along with reduced ROS production both secondary to improved mitochondrial function elicited by treatment with BEN.

Increased Expression of u-PA and u-PAR on Monocytes by LDL and Lp(a) Lipoproteins – Consequences for Plasmin Generation and Monocyte Adhesion

1999 ◽  
Vol 81 (04) ◽  
pp. 594-560 ◽  
Author(s):  
Florence Ganné ◽  
Marc Vasse ◽  
Jean-Louis Beaudeu ◽  
Jacqueline Peynet ◽  
Arnaud François ◽  
...  
Keyword(s):  
Fold Increase ◽  
Surface Expression ◽  
Cell Surface Expression ◽  
Atheromatous Plaque ◽  
Monocyte Adhesion ◽  
Blood Monocytes ◽  
Proteolytic Cascade ◽  
Ecm Degradation ◽  
Dose Dependent Increase ◽  
Dose Dependent

SummaryMonocyte-derived foam cells figure prominently in rupture-prone regions of atherosclerotic plaque. As urokinase/urokinase-receptor (u-PA/u-PAR) is the trigger of a proteolytic cascade responsible for ECM degradation, we have examined the effect of atherogenic lipoproteins on monocyte surface expression of u-PAR and u-PA. Peripheral blood monocytes, isolated from 10 healthy volunteers, were incubated with 10 to 200 µg/ml of native or oxidised (ox-) atherogenous lipoproteins for 18 h and cell surface expression of u-PA and u-PAR was analysed by flow cytometry. Both LDL and Lp(a) induced a dose-dependent increase in u-PA (1.6-fold increase with 200 μg/ml of ox-LDL) and u-PAR [1.7-fold increase with 200 μg/ml of ox-Lp(a)]. There is a great variability of the response among the donors, some of them remaining non-responders (absence of increase of u-PA or u-PAR) even at 200 μg/ml of lipoproteins. In positive responders, enhanced u-PA/u-PAR is associated with a significant increase of plasmin generation (1.9-fold increase with 200 μg/ml of ox-LDL), as determined by an amidolytic assay. Furthermore, monocyte adhesion to vitronectin and fibrinogen was significantly enhanced by the lipoproteins [respectively 2-fold and 1.7-fold increase with 200 μg/ml of ox-Lp(a)], due to the increase of u-PAR and ICAM-1, which are receptors for vitronectin and fibrinogen. These data suggest that atherogenous lipoproteins could contribute to the development of atheromatous plaque by increasing monocyte adhesion and trigger plaque weakening by inducing ECM degradation.

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