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NGAL Lacks Specificity for Acute Kidney Injury in Acute Heart Failure Syndrome
The 15th Annual Scientific Meeting survival rate was adjusted by the Kaplan-Meier. HF patients showed EF 3569%, 69.9% male, age 59613 years, 50.7% self-identified as black, 46% with ischemic etiology. The mean follow-up of 23 months showed 18 deaths and 46 hospitalizations. The genotype Glu27Glu (24.7 vs 6.1% p50.0004) and Arg16Arg (72.6 vs 22.8 p! 0.0001) of bAR2 polymorphisms were higher in HF patients compared with controls, as Gly49Gly (33.6 vs 4.3% p!0.0001) of bAR1 polymorphism. Patients with hospital admission showed Gly389 allelic frequency significantly higher (54.9% vs.42.1%, p50.039), and prevailed Gly389 allelic frequency among deaths (61.1%, p50.047). Black patients with genotype Ser49Ser showed a lower survival than to Gly49Gly homozygous and heterozygous Ser49Gly (p 5 0.028).
HFSA
S33
104 The Natriuretic Peptide Receptors NPR-A and NPR-B Are Differentially Regulation by Extracellular Matrix Proteins Brenda K. Huntley, Tomoko Ichiki, John C. Burnett Jr.; Cardiorenal Research, Mayo Clinic, Rochester, MN Background: Cardiac fibroblasts (CFs) regulate myocardial remodeling by proliferating, differentiating, and secreting extra-cellular matrix proteins (ECMs). Natriuretic peptides (NPs) are anti-fibrotic, inhibiting collagen production, augmenting matrix metalloproteinases, and suppressing CF proliferation. Recently, we demonstrated that the ECM proteins fibronectin (FN) and Collagen IV (Col IV) augmented production of BNP’s second messenger cGMP in CFs, supporting crosstalk between these ECM components and BNP involving its particulate guanylyl cyclase receptor, NPR-A. The goal of this investigation was to assess whether CNP and CNP-like peptides, which signal through the NPR-B receptor, are also affected by the ECM. Methods: Human CFs were cultured as previously described on ECM coated or non-coated plates. Cyclic GMP assays were done as previously reported. Results: As previously observed, BNP in vitro induced a dose dependent increase in cGMP which was augmented by FN and Col IV (p!0.05) that was abrogated by NPR-A antibody blockade. In contrast, activation of cGMP by two known active forms of CNP, CNP-22 and CNP-53 which bind to NPR-B, were not affected by ECM presence. Interestingly, the designer peptide CD-NP, known to signal through both NPR-A and NPR-B demonstrated an increase in cGMP with FN and Col IV similar to that observed with BNP. Conclusion: Our data suggests that NPR-A activation is augmented by ECM while NPR-B activation is not which may be important in understanding the physiological role of these cGMP receptors in the regulation of myocardial remodeling and guiding natriuretic peptide based therapeutics in its inhibition. Interestingly, the CNP-like chimeric in CF’s demonstrates properties of BNP supporting its unique role as an anti-fibrotic therapeutic.
105 Fig. Event-free survival according to the ADBR1 polymorphism ser 49Gly in black self-reported patients There was no association between improved LVEFO 20% and bAR polymorphisms. In conclusion Systolic HF outpatients with beta-blocker therapy, self-identified as black and homozygous Ser49Ser may have reduced event-free survival as such Gly389 carries, while Glu27Glu, Arg16Arg and Gly49Gly genotypes may be associated with risk for HF.
103 Chronic Treatment with Mineralocorticoid Receptor Antagonists Reverses SaltInduced Transcriptional Effects in Aorta of Dahl Salt Sensitive Rats Sheng Gao, Jun Li, Xiaoyan Zhou, Patricia Brown, Oscar Puig, Maarten Hoek, Michael Forrest, Sophie Roy, Martin Crook; Cardiovascular Diseases, MRL, Merck & Co., Rahway, NJ The mineralocorticoid receptor antagonists (MRAs) eplerenone and spironolactone are effective pharmacological therapies for hypertension and heart failure and are known to exert their therapeutic effects in multiple tissues including vascular tissue, but the transcriptional changes after chronic treatment are poorly defined. We sought to investigate the transcriptional effects in aorta of Dahl Salt Sensitive (Dahl SS) rats after 8 weeks of dosing with eplerenone (100mg/Kg/day in feed) on low salt (0.3%) and high salt (4%) diets by gene expression profiling. In vehicle groups, both systolic and diastolic BP significantly increased in rats on high salt diet. Eplerenone treatment significantly lowered BP in Dahl SS rats on both high salt and low salt diet. This was accompanied by reductions in heart and kidney weights and histopathological changes consistent with anti-fibrotic effects of MRAs. mRNA extracted from aorta of Dahl SS rats from all experimental groups were subjected to gene expression profiling. 1982 genes were differentially expressed between high salt and low salt groups. The treatment of rats on a high salt diet with eplerenone reversed 28% of the salt-induced expression change (552 out of 1982). Of these, 23 genes with jfold changejO1.7 were selected for further validation. Equivalent or greater transcriptional changes were observed in qRT-PCR analysis for 20 out of 23 genes, thus confirmed the finding from the profiling analysis. To further validate the gene list, we investigated the transcriptional changes of these selected genes in an independent study, in which Dahl SS rats on a high salt diet were dosed with vehicle or spironolactone (10, 30, and 100mg/Kg/day). 15 out of 22 salt-induced gene expression changes were significantly reversed by 100mg/Kg/day spironolactone. Three of those confirmed genes (LBP, CHI3L1, and PRG4) showed robust changes upon treatment of both MRAs and demonstrated dose-dependent responses to spironolactone. These three genes are involved in cell proliferation and inflammation, and may reflect some of the cardiovascular remodeling following MRA treatment. Furthermore, all three genes encode soluble proteins that could be used as biomarkers for vascular remodeling. These data highlight the pharmacodynamic effects of MRAs on the vasculature after chronic treatment and offer insight into the molecular mechanisms that underlie their therapeutic effects.
NGAL Lacks Specificity for Acute Kidney Injury in Acute Heart Failure Syndrome Mridula Rai1,2, Cara Statz1, Aleksandra Ras1, Jennifer Rahn1, Lynn O’Bara1, Faisal Zaeem1, Roja Mulamalla1, Jonathan Hammond1, Detlef Wencker1; 1Heart Failure and Transplant Center, Hartford Hospital, Hartford, CT; 2University of Connecticut Health Center, Farmington, CT Purpose: Neutrophil gelatinase-associated lipocalin (NGAL), a recently identified cytokine, is proposed as a novel marker of kidney injury in heart failure (HF). However, current studies have shown that an increased level of NGAL in the serum is a predictive marker of mortality in HF even in the absence of renal dysfunction, questioning its specificity for the kidney. Recently, a cardiac signaling pathway between interleukin 1 beta (IL-1) and NGAL expression has been identified in rat myocytes. IL-1 is known to be activated by its inflammasome, caspase 1 (C1), which is involved in the progression of cardiomyopathy. We seek to elicit the surge of NGAL in relation to C1 activity and define its role in end stage HF independent of its action in acute kidney injury. Methods and Materials: NGAL and C1 were assessed in different subsets of patients: decompensated HF (n537), cardiogenic shock (n525), and normal healthy controls (n54). Serum NGAL and C1 were measured in solid phase enzyme-linked immunosorbant assays (ELISA). Other variables including NYHA class, serum creatinine and serum BNP were also documented. Results: 62 patients (mean age 62614.4) with acute decompensated HF (ADHF) were studied. Twenty five patients were in cardiogenic shock and underwent ventricular assist device implant or heart transplant (VAD/TX). NGAL and C1 were markedly upregulated in ADHF patients compared to normal controls (p50.001 and !0.0001, respectively) and were significantly correlated to each other(R50.47, p50.003, n537). After VAD/TX, serum levels of C1 (p50.003) and NGAL (p50.04) decreased significantly. Of note, the rise of BNP in ADHF patients was inversely related with NGAL (R5 -0.48, p50.003) and C1 (R5 -0.19, p5NS) suggesting a direct biological effect of BNP on NGAL and C1. More importantly, serum creatinine level did not correlate with NGAL (R50.155, p50.36). Conclusions: During acute heart failure, the rise of NGAL occurs independent of creatinine. On the other hand, NGAL surge is associated with caspase 1 suggesting a systemic inflammatory response rather than acute kidney injury.
106 Circulating microRNA Patterns in Ischemic and Idiopathic Heart Failure Oxana Galenko1, Samin Panahi1, Staci Gunter1, Kimberly D. Brunisholz1, Benjamin D. Horne1,2, John Carlquist1,3, Jeffrey L. Anderson1,3; 1Cardiology, Intermountain Medical Center, Murray, UT; 2Genetic Epidemiology, University of Utah, Salt Lake City, UT; 3Cardiology, University of Utah, Salt Lake City, UT Introduction: MicroRNAs (miRNA) are short (w22 nucleotides) non-coding RNA molecules that inhibit translation of gene transcripts. MiRNAs create characteristic bio-signatures during development and under pathologic conditions. The aim of this study was to determine if peripheral blood miRNAs in newly diagnosed heart failure (HF) patients showed a unique pattern when compared to healthy individuals. Methods: Peripheral blood was obtained from consenting healthy controls (HC) and HF patients (n540), with either idiopathic (ID) or ischemic (IS) cardiomyopathy.
HFSA
S33
104 The Natriuretic Peptide Receptors NPR-A and NPR-B Are Differentially Regulation by Extracellular Matrix Proteins Brenda K. Huntley, Tomoko Ichiki, John C. Burnett Jr.; Cardiorenal Research, Mayo Clinic, Rochester, MN Background: Cardiac fibroblasts (CFs) regulate myocardial remodeling by proliferating, differentiating, and secreting extra-cellular matrix proteins (ECMs). Natriuretic peptides (NPs) are anti-fibrotic, inhibiting collagen production, augmenting matrix metalloproteinases, and suppressing CF proliferation. Recently, we demonstrated that the ECM proteins fibronectin (FN) and Collagen IV (Col IV) augmented production of BNP’s second messenger cGMP in CFs, supporting crosstalk between these ECM components and BNP involving its particulate guanylyl cyclase receptor, NPR-A. The goal of this investigation was to assess whether CNP and CNP-like peptides, which signal through the NPR-B receptor, are also affected by the ECM. Methods: Human CFs were cultured as previously described on ECM coated or non-coated plates. Cyclic GMP assays were done as previously reported. Results: As previously observed, BNP in vitro induced a dose dependent increase in cGMP which was augmented by FN and Col IV (p!0.05) that was abrogated by NPR-A antibody blockade. In contrast, activation of cGMP by two known active forms of CNP, CNP-22 and CNP-53 which bind to NPR-B, were not affected by ECM presence. Interestingly, the designer peptide CD-NP, known to signal through both NPR-A and NPR-B demonstrated an increase in cGMP with FN and Col IV similar to that observed with BNP. Conclusion: Our data suggests that NPR-A activation is augmented by ECM while NPR-B activation is not which may be important in understanding the physiological role of these cGMP receptors in the regulation of myocardial remodeling and guiding natriuretic peptide based therapeutics in its inhibition. Interestingly, the CNP-like chimeric in CF’s demonstrates properties of BNP supporting its unique role as an anti-fibrotic therapeutic.
105 Fig. Event-free survival according to the ADBR1 polymorphism ser 49Gly in black self-reported patients There was no association between improved LVEFO 20% and bAR polymorphisms. In conclusion Systolic HF outpatients with beta-blocker therapy, self-identified as black and homozygous Ser49Ser may have reduced event-free survival as such Gly389 carries, while Glu27Glu, Arg16Arg and Gly49Gly genotypes may be associated with risk for HF.
103 Chronic Treatment with Mineralocorticoid Receptor Antagonists Reverses SaltInduced Transcriptional Effects in Aorta of Dahl Salt Sensitive Rats Sheng Gao, Jun Li, Xiaoyan Zhou, Patricia Brown, Oscar Puig, Maarten Hoek, Michael Forrest, Sophie Roy, Martin Crook; Cardiovascular Diseases, MRL, Merck & Co., Rahway, NJ The mineralocorticoid receptor antagonists (MRAs) eplerenone and spironolactone are effective pharmacological therapies for hypertension and heart failure and are known to exert their therapeutic effects in multiple tissues including vascular tissue, but the transcriptional changes after chronic treatment are poorly defined. We sought to investigate the transcriptional effects in aorta of Dahl Salt Sensitive (Dahl SS) rats after 8 weeks of dosing with eplerenone (100mg/Kg/day in feed) on low salt (0.3%) and high salt (4%) diets by gene expression profiling. In vehicle groups, both systolic and diastolic BP significantly increased in rats on high salt diet. Eplerenone treatment significantly lowered BP in Dahl SS rats on both high salt and low salt diet. This was accompanied by reductions in heart and kidney weights and histopathological changes consistent with anti-fibrotic effects of MRAs. mRNA extracted from aorta of Dahl SS rats from all experimental groups were subjected to gene expression profiling. 1982 genes were differentially expressed between high salt and low salt groups. The treatment of rats on a high salt diet with eplerenone reversed 28% of the salt-induced expression change (552 out of 1982). Of these, 23 genes with jfold changejO1.7 were selected for further validation. Equivalent or greater transcriptional changes were observed in qRT-PCR analysis for 20 out of 23 genes, thus confirmed the finding from the profiling analysis. To further validate the gene list, we investigated the transcriptional changes of these selected genes in an independent study, in which Dahl SS rats on a high salt diet were dosed with vehicle or spironolactone (10, 30, and 100mg/Kg/day). 15 out of 22 salt-induced gene expression changes were significantly reversed by 100mg/Kg/day spironolactone. Three of those confirmed genes (LBP, CHI3L1, and PRG4) showed robust changes upon treatment of both MRAs and demonstrated dose-dependent responses to spironolactone. These three genes are involved in cell proliferation and inflammation, and may reflect some of the cardiovascular remodeling following MRA treatment. Furthermore, all three genes encode soluble proteins that could be used as biomarkers for vascular remodeling. These data highlight the pharmacodynamic effects of MRAs on the vasculature after chronic treatment and offer insight into the molecular mechanisms that underlie their therapeutic effects.
NGAL Lacks Specificity for Acute Kidney Injury in Acute Heart Failure Syndrome Mridula Rai1,2, Cara Statz1, Aleksandra Ras1, Jennifer Rahn1, Lynn O’Bara1, Faisal Zaeem1, Roja Mulamalla1, Jonathan Hammond1, Detlef Wencker1; 1Heart Failure and Transplant Center, Hartford Hospital, Hartford, CT; 2University of Connecticut Health Center, Farmington, CT Purpose: Neutrophil gelatinase-associated lipocalin (NGAL), a recently identified cytokine, is proposed as a novel marker of kidney injury in heart failure (HF). However, current studies have shown that an increased level of NGAL in the serum is a predictive marker of mortality in HF even in the absence of renal dysfunction, questioning its specificity for the kidney. Recently, a cardiac signaling pathway between interleukin 1 beta (IL-1) and NGAL expression has been identified in rat myocytes. IL-1 is known to be activated by its inflammasome, caspase 1 (C1), which is involved in the progression of cardiomyopathy. We seek to elicit the surge of NGAL in relation to C1 activity and define its role in end stage HF independent of its action in acute kidney injury. Methods and Materials: NGAL and C1 were assessed in different subsets of patients: decompensated HF (n537), cardiogenic shock (n525), and normal healthy controls (n54). Serum NGAL and C1 were measured in solid phase enzyme-linked immunosorbant assays (ELISA). Other variables including NYHA class, serum creatinine and serum BNP were also documented. Results: 62 patients (mean age 62614.4) with acute decompensated HF (ADHF) were studied. Twenty five patients were in cardiogenic shock and underwent ventricular assist device implant or heart transplant (VAD/TX). NGAL and C1 were markedly upregulated in ADHF patients compared to normal controls (p50.001 and !0.0001, respectively) and were significantly correlated to each other(R50.47, p50.003, n537). After VAD/TX, serum levels of C1 (p50.003) and NGAL (p50.04) decreased significantly. Of note, the rise of BNP in ADHF patients was inversely related with NGAL (R5 -0.48, p50.003) and C1 (R5 -0.19, p5NS) suggesting a direct biological effect of BNP on NGAL and C1. More importantly, serum creatinine level did not correlate with NGAL (R50.155, p50.36). Conclusions: During acute heart failure, the rise of NGAL occurs independent of creatinine. On the other hand, NGAL surge is associated with caspase 1 suggesting a systemic inflammatory response rather than acute kidney injury.
106 Circulating microRNA Patterns in Ischemic and Idiopathic Heart Failure Oxana Galenko1, Samin Panahi1, Staci Gunter1, Kimberly D. Brunisholz1, Benjamin D. Horne1,2, John Carlquist1,3, Jeffrey L. Anderson1,3; 1Cardiology, Intermountain Medical Center, Murray, UT; 2Genetic Epidemiology, University of Utah, Salt Lake City, UT; 3Cardiology, University of Utah, Salt Lake City, UT Introduction: MicroRNAs (miRNA) are short (w22 nucleotides) non-coding RNA molecules that inhibit translation of gene transcripts. MiRNAs create characteristic bio-signatures during development and under pathologic conditions. The aim of this study was to determine if peripheral blood miRNAs in newly diagnosed heart failure (HF) patients showed a unique pattern when compared to healthy individuals. Methods: Peripheral blood was obtained from consenting healthy controls (HC) and HF patients (n540), with either idiopathic (ID) or ischemic (IS) cardiomyopathy.