Abstract 16892: Beneficial Effect of miR-25 Decoy Overexpression in a Murine Model of Heart Failure

Introduction: Recently, our group found that miR-25 is a key microRNA that regulates SERCA2a and we showed anti-miR25 treatment enhanced cardiac contractility and function through SERCA2a restoration in murine heart failure model. However, the strong and stable suppression of specific microRNA activity would be essential to evaluate the therapeutic potential of such an approach. In this study, we constructed a miR-25 decoy t using TuD RNA (tough decoy RNAs) and miR-25 decoy activity was evaluated in various settings. Methods: First, miR-25 decoy construct was applied to both isolated adult cardiomyocytes and H9C2 cardiomyoblasts. A pMirTarget vector containing SERCA2a 3’-UTR under luciferase was used to evaluate this decoy specificity. Second, we generated pressure-overload heart failure models in mouse by TAC operation. In the HF mice, AAV9-GFP (control) and AAV9-miR-25 decoy were delivered by tail vein injection. Two months following gene transfer, cardiac function was evaluated by echocardiography and invasive hemodynamics. Protein and RNA expression levels of SERCA2a and miR-25 expression level were confirmed by qRT-PCR analysis. Results: First, we observed that we were able to achieve about 2-3 fold increase of SERCA2a expression by miR-25 decoy transfection in both H9C2 and in isolated adult cardiomyocytes. Also, similar expression pattern was confirmed in the heart of miR-25 decoy injected normal mice. Second, the HF model by TAC surgery was confirmed with echocardiography. Overall average FS (%) in HF was 37.77+/- 8.75 (n=10) and in non-surgery control mice was 62.51 +/- 3.42(n=4). After AAV9 injection, cardiac function of AAV9-miR-25 decoy injected mice was enhanced but AAV9-GFP injected mice showed severe cardiac dysfunction and dilation (AAV-GFP (n=6) vs AAV-miR-25 decoy (n=4)). Third, western blot analysis showed that SERCA2a expression was significantly restored in miR-25 decoy injected mice. In addition, we confirmed that miR-25 expression level was kept low by qRT-PCR analysis. Taken together, our data would indicate that using miR-25 decoy is an effective strategy for the long-term suppression of miR-25 and it may be a promising therapeutic target to restore SERCA2a and reverse the HF phenotype.

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Abstract 381: Mcur1 is a Potential Target to Modulate Cardiac Contractility and Alleviate Cardiac Function During Heart Failure

Circulation Research ◽

10.1161/res.127.suppl_1.381 ◽

2020 ◽

Vol 127 (Suppl_1) ◽

Author(s):

Rajika Roy ◽

Santhanam Shanmughapriya ◽

Xueqian Zhang ◽

Jianliang Song ◽

Dhanendra Tomar ◽

...

Keyword(s):

Heart Failure ◽

Cardiac Function ◽

Contractile Function ◽

Cardiac Contractility ◽

Pressure Overload ◽

Dominant Negative ◽

Selective Channel

Cardiac contractility is regulated by the intracellular Ca 2+ concentration fluxes which are actively regulated by multiple channels and transporters. Ca 2+ uptake into the mitochondrial matrix is precisely controlled by the highly Ca 2+ selective channel, Mitochondrial Calcium Uniporter (MCU). Earlier studies on the cardiac-specific acute MCU knockout and a transgenic dominant-negative MCU mice have demonstrated that mitochondrial Ca 2+ ( m Ca 2+ ) signaling is necessary for cardiac ‘‘fight-or-flight’’ contractile response, however, the role of m Ca 2+ buffering to shape global cytosolic Ca 2+ levels and affect E-C coupling, particularly the Ca 2+ transient, on a beat-to-beat basis still remains to be solved. Our earlier studies have demonstrated that loss of MCU Regulator 1 (MCUR1) in cardiomyocytes results in the impaired m Ca 2+ uptake. We have now employed the cardiac-specific MCUR1 knockout mouse to dissect the precise role of MCU in regulating cytosolic Ca 2+ transients associated with excitation-contraction (E-C) coupling and cardiac function. Results from our studies including the in vivo analyses of cardiac physiology during normal and pressure-overloaded mouse models and in vitro experiments including single-cell cardiac contractility, calcium transients, and electrophysiology measurements demonstrate that MCUR1/MCU regulated m Ca 2+ buffering in cardiomyocytes, although insignificant under basal condition, becomes critical in stress induced conditions and actively participates in regulating the c Ca 2+ transients. Also, the ablation of MCUR1 in cardiomyocytes during stress conditions prevents m Ca 2+ overload and subsequent mROS overproduction. Our data indicate that MCUR1 ablation offers protection against pressure-overload cardiac hypertrophy. In summary, our results provide critical insights into the mechanisms by which the MCU channel contributes in regulating the contractile function of the cardiomyocytes and the role of m Ca 2+ in the development and progression of heart failure.

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Investigation of olfactory receptor family 51 subfamily j member 1 (OR51J1) gene susceptibility as a potential breast cancer-associated biomarker

PLoS ONE ◽

10.1371/journal.pone.0246752 ◽

2021 ◽

Vol 16 (2) ◽

pp. e0246752

Author(s):

Maryam Asadi ◽

Nahid Ahmadi ◽

Simin Ahmadvand ◽

Ali Akbar Jafari ◽

Akbar Safaei ◽

...

Keyword(s):

Breast Cancer ◽

Statistical Analysis ◽

Protein Expression ◽

Olfactory Receptor ◽

Mrna Level ◽

Expression Level ◽

Pcr Analysis ◽

Normal Tissues ◽

Qrt Pcr ◽

Human Protein Atlas

Among cancer treatment methods, targeted therapy using cancer-associated biomarkers has minimum side effects. Recently olfactory receptor (OR) family attracts the researcher’s attention as a favorable biomarker of cancer. Here, a statistical approach using complete data from the human protein atlas database was used to evaluate the potential of OR51J1 gene as a cancer-associated biomarker. To confirm the findings of statistical analysis, the OR51J1 mRNA and protein expression levels in breast tumor and normal tissue were measured using quantitative Real Time PCR (qRT-PCR) and immunohistochemistry (IHC) techniques. The association with clinicopathological factors was analyzed. Statistical analysis revealed that OR51J1 has a high expression level in more than 20 types of cancer tissues without any expression in 44 normal tissues. In 15 cancer types, including breast cancer, expression score was more than 90%. The qRT-PCR analysis in breast cancer showed OR51J1 have significantly higher expression level in tumors than normal tissues (2.91 fold). The IHC results showed OR51J1 expression on other cellular subtypes than tumor and normal cells, including myoepithelium, fibroblast, and lymphocytes. OR51J1 protein expression in invasive cells, as well as its overall score, showed a significant correlation with ER and PR expression and breast cancer (BC) subtypes. Results revealed the potential of OR51J1 as a cancer-associated biomarker for the diagnosis of breast cancer at the mRNA level.

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A gene therapeutic approach to inhibit calcium and integrin binding protein 1 ameliorates maladaptive remodelling in pressure overload

Cardiovascular Research ◽

10.1093/cvr/cvy154 ◽

2018 ◽

Vol 115 (1) ◽

pp. 71-82 ◽

Cited By ~ 5

Author(s):

Andrea Grund ◽

Malgorzata Szaroszyk ◽

Janina K Döppner ◽

Mona Malek Mohammadi ◽

Badder Kattih ◽

...

Keyword(s):

Heart Failure ◽

Cardiac Hypertrophy ◽

Cardiac Function ◽

Binding Protein ◽

Treatment Strategies ◽

Pressure Overload ◽

Cellular Growth ◽

Neonatal Rat ◽

Rat Cardiomyocytes ◽

Pathological Cardiac Hypertrophy

Abstract Aims Chronic heart failure is becoming increasingly prevalent and is still associated with a high mortality rate. Myocardial hypertrophy and fibrosis drive cardiac remodelling and heart failure, but they are not sufficiently inhibited by current treatment strategies. Furthermore, despite increasing knowledge on cardiomyocyte intracellular signalling proteins inducing pathological hypertrophy, therapeutic approaches to target these molecules are currently unavailable. In this study, we aimed to establish and test a therapeutic tool to counteract the 22 kDa calcium and integrin binding protein (CIB) 1, which we have previously identified as nodal regulator of pathological cardiac hypertrophy and as activator of the maladaptive calcineurin/NFAT axis. Methods and results Among three different sequences, we selected a shRNA construct (shCIB1) to specifically down-regulate CIB1 by 50% upon adenoviral overexpression in neonatal rat cardiomyocytes (NRCM), and upon overexpression by an adeno-associated-virus (AAV) 9 vector in mouse hearts. Overexpression of shCIB1 in NRCM markedly reduced cellular growth, improved contractility of bioartificial cardiac tissue and reduced calcineurin/NFAT activation in response to hypertrophic stimulation. In mice, administration of AAV-shCIB1 strongly ameliorated eccentric cardiac hypertrophy and cardiac dysfunction during 2 weeks of pressure overload by transverse aortic constriction (TAC). Ultrastructural and molecular analyses revealed markedly reduced myocardial fibrosis, inhibition of hypertrophy associated gene expression and calcineurin/NFAT as well as ERK MAP kinase activation after TAC in AAV-shCIB1 vs. AAV-shControl treated mice. During long-term exposure to pressure overload for 10 weeks, AAV-shCIB1 treatment maintained its anti-hypertrophic and anti-fibrotic effects, but cardiac function was no longer improved vs. AAV-shControl treatment, most likely resulting from a reduction in myocardial angiogenesis upon downregulation of CIB1. Conclusions Inhibition of CIB1 by a shRNA-mediated gene therapy potently inhibits pathological cardiac hypertrophy and fibrosis during pressure overload. While cardiac function is initially improved by shCIB1, this cannot be kept up during persisting overload.

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Distinct roles of resident and nonresident macrophages in nonischemic cardiomyopathy

Proceedings of the National Academy of Sciences ◽

10.1073/pnas.1720065115 ◽

2018 ◽

Vol 115 (20) ◽

pp. E4661-E4669 ◽

Cited By ~ 35

Author(s):

Xudong Liao ◽

Yuyan Shen ◽

Rongli Zhang ◽

Keiki Sugi ◽

Neelakantan T. Vasudevan ◽

...

Keyword(s):

Heart Failure ◽

Cardiac Function ◽

Late Phase ◽

Myeloid Cells ◽

Pressure Overload ◽

Genetic Mutations ◽

Nonischemic Cardiomyopathy ◽

Resident Macrophage ◽

Temporal And Spatial

Nonischemic cardiomyopathy (NICM) resulting from long-standing hypertension, valvular disease, and genetic mutations is a major cause of heart failure worldwide. Recent observations suggest that myeloid cells can impact cardiac function, but the role of tissue-intrinsic vs. tissue-extrinsic myeloid cells in NICM remains poorly understood. Here, we show that cardiac resident macrophage proliferation occurs within the first week following pressure overload hypertrophy (POH; a model of heart failure) and is requisite for the heart’s adaptive response. Mechanistically, we identify Kruppel-like factor 4 (KLF4) as a key transcription factor that regulates cardiac resident macrophage proliferation and angiogenic activities. Finally, we show that blood-borne macrophages recruited in late-phase POH are detrimental, and that blockade of their infiltration improves myocardial angiogenesis and preserves cardiac function. These observations demonstrate previously unappreciated temporal and spatial roles for resident and nonresident macrophages in the development of heart failure.

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GW28-e0941 Increasing fatty acid utilization improves cardiac function through OPA1-mediated mitochondrial fusion in pressure overload-induced heart failure

Journal of the American College of Cardiology ◽

10.1016/j.jacc.2017.07.225 ◽

2017 ◽

Vol 70 (16) ◽

pp. C66

Author(s):

Yongzheng Guo ◽

Xing Zhang ◽

Feng Gao

Keyword(s):

Heart Failure ◽

Fatty Acid ◽

Cardiac Function ◽

Pressure Overload ◽

Mitochondrial Fusion ◽

Fatty Acid Utilization

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Osteopontin RNA aptamer can prevent and reverse pressure overload-induced heart failure

Cardiovascular Research ◽

10.1093/cvr/cvx016 ◽

2017 ◽

Vol 113 (6) ◽

pp. 633-643 ◽

Cited By ~ 21

Author(s):

Jihe Li ◽

Keyvan Yousefi ◽

Wen Ding ◽

Jayanti Singh ◽

Lina A. Shehadeh

Keyword(s):

Heart Failure ◽

Cardiac Function ◽

Cardiac Dysfunction ◽

Cardiac Fibrosis ◽

Cardiac Myocyte ◽

Pressure Overload ◽

Cardiomyocyte Hypertrophy ◽

Therapeutic Effects ◽

Rna Aptamer ◽

Myocyte Hypertrophy

Aims Cardiac myocyte hypertrophy, the main compensatory response to chronic stress in the heart often progresses to a state of decompensation that can lead to heart failure. Osteopontin (OPN) is an effector for extracellular signalling that induces myocyte growth and fibrosis. Although increased OPN activity has been observed in stressed myocytes and fibroblasts, the detailed and long term effects of blocking OPN signalling on the heart remain poorly defined. Targeting cardiac OPN protein by an RNA aptamer may be beneficial for tuning down OPN pathologic signalling. We aimed to demonstrate the therapeutic effects of an OPN RNA aptamer on cardiac dysfunction. Methods and results In vivo, we show that in a mouse model of pressure overload, treating at the time of surgeries with an OPN aptamer prevented cardiomyocyte hypertrophy and cardiac fibrosis, blocked OPN downstream signalling (PI3K and Akt phosphorylation), reduced expression of extracellular matrix (Lum, Col3a1, Fn1) and hypertrophy (Nppa, Nppb) genes, and prevented cardiac dysfunction. Treating at two months post-surgeries with the OPN aptamer reversed cardiac dysfunction and fibrosis and myocyte hypertrophy. While genetic homozygous deletion of OPN reduced myocardial wall thickness, surprisingly cardiac function and myocardial fibrosis, specifically collagen deposition and myofibroblast infiltration, were worse compared with wild type mice at three months of pressure overload. Conclusion Taken together, these data demonstrate that tuning down cardiac OPN signalling by an OPN RNA aptamer is a novel and effective approach for preventing cardiac hypertrophy and fibrosis, improving cardiac function, and reversing pressure overload-induced heart failure.

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Growth hormone-releasing hormone attenuates cardiac hypertrophy and improves heart function in pressure overload-induced heart failure

Proceedings of the National Academy of Sciences ◽

10.1073/pnas.1712612114 ◽

2017 ◽

Vol 114 (45) ◽

pp. 12033-12038 ◽

Cited By ~ 17

Author(s):

Iacopo Gesmundo ◽

Michele Miragoli ◽

Pierluigi Carullo ◽

Letizia Trovato ◽

Veronica Larcher ◽

...

Keyword(s):

Heart Failure ◽

Growth Hormone ◽

Cardiac Hypertrophy ◽

Cardiac Function ◽

Therapeutic Potential ◽

Growth Hormone Releasing Hormone ◽

Transverse Aortic Constriction ◽

Aortic Constriction

It has been shown that growth hormone-releasing hormone (GHRH) reduces cardiomyocyte (CM) apoptosis, prevents ischemia/reperfusion injury, and improves cardiac function in ischemic rat hearts. However, it is still not known whether GHRH would be beneficial for life-threatening pathological conditions, like cardiac hypertrophy and heart failure (HF). Thus, we tested the myocardial therapeutic potential of GHRH stimulation in vitro and in vivo, using GHRH or its agonistic analog MR-409. We show that in vitro, GHRH(1-44)NH2 attenuates phenylephrine-induced hypertrophy in H9c2 cardiac cells, adult rat ventricular myocytes, and human induced pluripotent stem cell-derived CMs, decreasing expression of hypertrophic genes and regulating hypertrophic pathways. Underlying mechanisms included blockade of Gq signaling and its downstream components phospholipase Cβ, protein kinase Cε, calcineurin, and phospholamban. The receptor-dependent effects of GHRH also involved activation of Gαs and cAMP/PKA, and inhibition of increase in exchange protein directly activated by cAMP1 (Epac1). In vivo, MR-409 mitigated cardiac hypertrophy in mice subjected to transverse aortic constriction and improved cardiac function. Moreover, CMs isolated from transverse aortic constriction mice treated with MR-409 showed improved contractility and reversal of sarcolemmal structure. Overall, these results identify GHRH as an antihypertrophic regulator, underlying its therapeutic potential for HF, and suggest possible beneficial use of its analogs for treatment of pathological cardiac hypertrophy.

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Chronic Benzene Exposure Aggravates Pressure Overload-Induced Cardiac Dysfunction

10.1101/2021.08.31.458367 ◽

2021 ◽

Author(s):

Igor N Zelko ◽

Sujith Dassanayaka ◽

Marina V Malovichko ◽

Caitlin M Howard ◽

Lauren F Garrett ◽

...

Keyword(s):

Heart Failure ◽

Endothelial Cells ◽

Cardiac Function ◽

Cardiac Tissue ◽

Pressure Overload ◽

Endothelial Activation ◽

Air Exposure ◽

Benzene Exposure ◽

Increased Risk

Benzene is a ubiquitous environmental pollutant abundant in household products, petrochemicals and cigarette smoke. Benzene is a well-known carcinogen in humans and experimental animals; however, little is known about the cardiovascular toxicity of benzene. Recent population-based studies indicate that benzene exposure is associated with an increased risk for heart failure. Nonetheless, it is unclear whether benzene exposure is sufficient to induce and/or exacerbate heart failure. We examined the effects of benzene (50 ppm, 6 h/day, 5 days/week, 6 weeks) or HEPA-filtered air exposure on transverse aortic constriction (TAC)-induced pressure overload in male C57BL/6J mice. Our data show that benzene exposure had no effect on cardiac function in the Sham group; however, it significantly compromised cardiac function as depicted by a significant decrease in fractional shortening and ejection fraction, as compared with TAC/Air-exposed mice. RNA-seq analysis of the cardiac tissue from the TAC/benzene-exposed mice showed a significant increase in several genes associated with adhesion molecules, cell-cell adhesion, inflammation, and stress response. In particular, neutrophils were implicated in our unbiased analyses. Indeed, immunofluorescence studies showed that TAC/benzene exposure promotes infiltration of CD11b+/S100A8+/myeloperoxidase+-positive neutrophils in the hearts by 3-fold. In vitro, the benzene metabolites, hydroquinone and catechol, induced the expression of P-selectin in cardiac microvascular endothelial cells by 5-fold and increased the adhesion of neutrophils to these endothelial cells by 1.5-2.0-fold. Benzene metabolite-induced adhesion of neutrophils to the endothelial cells was attenuated by anti-P-selectin antibody. Together, these data suggest that benzene exacerbates heart failure by promoting endothelial activation and neutrophil recruitment.

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Valsartan Regulates PI3K/AKT Pathways through lncRNA GASL1 to Improve Isoproterenol-Induced Heart Failure

Disease Markers ◽

10.1155/2022/1447399 ◽

2022 ◽

Vol 2022 ◽

pp. 1-8

Author(s):

Jian Zhou ◽

Xiujuan Duan ◽

Jibing Wang ◽

Yunhong Feng ◽

Jiangyong Yuan

Keyword(s):

Heart Failure ◽

Cardiac Function ◽

Akt Pathway ◽

Chain Reaction ◽

Qrt Pcr ◽

Polymerase Chain ◽

And Function ◽

Rat Tail

Objective. This study is aimed at determining the expression and function of the GASL1 and PI3K/AKT pathways in isoproterenol- (ISO-) induced heart failure (HF). To determine the moderating effect of valsartan (VAL) on the progression of ISO-induced HF and to elucidate the related mechanism. Materials and Methods. First, in in vivo experiment, we examined the effect of VAL on cardiac function in rats with ISO-induced HF. Similarly, quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot were used to detect the effect of VAL on ISO-treated rat primary cardiomyocytes. Then, si-GASL1-transfected primary cardiomyocytes were constructed and Ad-si-GASL1 was injected through rat tail vein to achieve the effect of lowering GASL1 expression, so as to investigate the role of GASL1 in VAL’s treatment of ISO-induced HF. Results. In ISO-induced HF rat models, the GASL1 decreased while PI3K and p-AKT expressions were abnormally elevated and cardiac function deteriorated, and VAL was able to reverse these changes. In primary cardiomyocytes, ISO induces apoptosis of cardiomyocytes, and expression of GASL1 decreased while PI3K and p-AKT were abnormally elevated, which can be reversed by VAL. The transfection of primary cardiomyocytes with si-GASL1 confirmed that GASL1 affected the expression of PI3K, p-AKT, and the apoptosis of primary cardiomyocytes. Rat myocardium injected with Ad-si-GASL1 was found to aggravate the cardiac function improved by VAL. Conclusions. This study was the first to confirm that VAL improves ISO-induced HF by regulating the PI3K/AKT pathway through GASL1. And this study demonstrated a significant correlation between HF, VAL, GASL1, and the PI3K/AKT pathway.

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