Specific Aims: Lack of motivation to exercise is the hallmark of obesity and type2 diabetes (T2D). High fat diet (HFD) is the major cause of obesity and T2D that impair contractility of cardiomyocytes leading to heart failure. The stimulation of 2-adrenergic receptors (2-AR) induces contractility and mitigates cardiac dysfunction. Elevated level of homocysteine (Hcy) called hyperhomocysteinemia (HHcy, a co-morbid condition) is associated with heart failure in diabetes. Hcy competes with and antagonizes 2-AR. The long term goal of the project is to understand the role of 2- AR and Hcy axis in diabetes. HHcy is ameliorated by conversion of Hcy to H2S (an anti-hypertensive, vasorelaxing, and anti-oxidant gas) by cystathionine synthase (CBS) and cystathionine ? lyase (CSE). In diabetes, the levels of CBS and CSE are suppressed that induces HHcy and thereby impairs formation of H2S. H2S is an inducer of AKT (an anti-oxidant involved in glucose metabolism) and AKT is down regulated in diabetes. We have shown that H2S induces a stimulatory G-protein couple receptor called Gs (inducer of 2-AR) in diabetic cardiomyocytes. On the other hand, HHcy attenuates Gs. However, exercise mitigates HHcy and up regulates 2-AR in T2D. Also, exercise and salbutamol (2-AR agonist) have synergistic effect on mitigation of contractile dysfunction in diabetic cardiomyocytes. However, the underlying mechanism is unclear. We have shown that in diabetes, HHcy activates MMP9 that induces cardiac fibrosis and impairs myocardial contractility. Also, microRNA-133a (miR-133a) that regulates cardiac fibrosis and hypertrophy is attenuated in diabetic hearts. Our preliminary studies show that HHcy (CBS+/- mice) and HFD treatment attenuate myocardial 2-AR and miR-133a, whereas exercise and H2S mitigate this attenuation. Exercise and salbutamol have synergistic effect on induction of AKT and Gs in diabetes. The transgenic expression of 2-AR (2-ARTg) mitigates HFD induced cardiac fibrosis. Also, ablation of MMP9 gene ameliorates HFD induced contractile dysfunction. Both HFD treated WT and CBS+/- mice have left ventricle dysfunction and impaired contractility, which is mitigated by exercise training. HHcy and HFD also down regulates sarco-endoplasmic reticulum ATPase 2a (serca2a; regulates calcium flux during muscle contraction), which is improved by exercise and H2S. The central hypothesis of the proposal is that in HFD induced diabetes, the intolerance to exercise is, in part, due to attenuation of 2-AR and Gs by HHcy that down regulates AKT and miR-133a and induces MMP9 causing myocytes contractile dysfunction. The exercise and H2S induce 2-AR and Gs by mitigating HHcy and ameliorate contractile dysfunction in diabetes (Figure 1). We will test this hypothesis by following three specific aims:
Specific Aim#1 : To determine whether the 2-AR is attenuated by hyperhomocysteinemia in diabetes and exercise and H2S mitigate this attenuation. Hypothesis: In diabetes, 2-AR is down regulated and Hcy is elevated, and exercise and H2S induce 2-AR and decrease Hcy level.
Specific Aim#2 : To determine whether the AKT and miR-133a are attenuated and MMP9 is induced by hyperhomocysteinemia in diabetes and exercise and H2S ameliorate this attenuation and induction. Hypothesis: In diabetes, the elevated level of Hcy inhibits AKT and miR-133a and induces MMP9. Exercise and H2S induce AKT and attenuate MMP9 by decreasing Hcy level in diabetic hearts.
Specific Aim#3 : To determine whether the contractility of cardiomyocytes is impaired by hyperhomocysteinemia in diabetes and exercise and H2S mitigate the contractile dysfunction. Hypothesis: In diabetes, Hcy and MMP9 are elevated while 2-AR and AKT are attenuated leading to contractile dysfunction, and exercise and H2S ameliorate these levels and mitigate contractile dysfunction. These studies will elucidate the mechanism of homocysteine-mediated attenuation of 2-AR in diabetes and cardio-protective role of 2-AR agonist, exercise and H2S in diabetic complications.

Public Health Relevance

Obesity mediated type 2diabetes (T2D) are leading problems in the western world and the rate of diabetes patients are increasing rapidly across the globe. However, the cause is not completely comprehended. It is found that diabetes increases the chances of heart failure and homocysteine levels is elevated in diabetes and heart failure conditions, but how homocysteine affect heart failure in diabetes is not clear. We have reported that 2-adrenergic receptors (2-AR) play crucial role in improving the contractility of heart and is attenuated in diabetic condition. However, how homocysteine interacts with 2-AR in diabetic hearts is not known. In this study, we propose that elevated level of homocysteine down regulates 2-AR by inhibiting its stimulator (Gs) and competing for their binding sites. Using different strains of mice where homocysteine and 2-AR levels are elevated, we demonstrated that homocysteine cross talk with 2-AR. We also investigated the mechanism of homocysteine mediated attenuation of 2-AR and its effect on contractility of heart and how exercise and hydrogen sulfide (a cardioprotective gas) improves 2-AR in obese and diabetic mice. These findings provide a novel mechanism of homocysteine mediated contractile dysfunction in diabetic hearts and effect of exercise and H2S in amelioration of contractile dysfunction.

National Institute of Health (NIH)
National Heart, Lung, and Blood Institute (NHLBI)
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Special Emphasis Panel (ZRG1)
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Adhikari, Bishow B
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University of Nebraska Medical Center
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Yadav, Santosh Kumar; Mishra, Paras Kumar (2018) Isolation, Characterization and Differentiation of Mouse Cardiac Progenitor Cells. Methods Mol Biol 1842:183-191
Nandi, Shyam Sundar; Shahshahan, Hamid Reza; Shang, Quanliang et al. (2018) MiR-133a Mimic Alleviates T1DM-Induced Systolic Dysfunction in Akita: An MRI-Based Study. Front Physiol 9:1275
Nandi, Shyam Sundar; Mishra, Paras Kumar (2018) Targeting miRNA for Therapy of Juvenile and Adult Diabetic Cardiomyopathy. Adv Exp Med Biol 1056:47-59
Prathipati, Priyanka; Nandi, Shyam Sundar; Mishra, Paras Kumar (2017) Stem Cell-Derived Exosomes, Autophagy, Extracellular Matrix Turnover, and miRNAs in Cardiac Regeneration during Stem Cell Therapy. Stem Cell Rev 13:79-91
Mishra, Paras K; Ying, Wei; Nandi, Shyam Sundar et al. (2017) Diabetic Cardiomyopathy: An Immunometabolic Perspective. Front Endocrinol (Lausanne) 8:72
Nandi, Shyam Sundar; Mishra, Paras Kumar (2017) H2S and homocysteine control a novel feedback regulation of cystathionine beta synthase and cystathionine gamma lyase in cardiomyocytes. Sci Rep 7:3639
Kesherwani, Varun; Shahshahan, Hamid R; Mishra, Paras K (2017) Cardiac transcriptome profiling of diabetic Akita mice using microarray and next generation sequencing. PLoS One 12:e0182828
Nandi, Shyam Sundar; Zheng, Hong; Sharma, Neeru M et al. (2016) Lack of miR-133a Decreases Contractility of Diabetic Hearts: A Role for Novel Cross Talk Between Tyrosine Aminotransferase and Tyrosine Hydroxylase. Diabetes 65:3075-90
Hackfort, Bryan T; Mishra, Paras K (2016) Emerging role of hydrogen sulfide-microRNA crosstalk in cardiovascular diseases. Am J Physiol Heart Circ Physiol 310:H802-12
Prathipati, Priyanka; Metreveli, Naira; Nandi, Shyam Sundar et al. (2016) Ablation of Matrix Metalloproteinase-9 Prevents Cardiomyocytes Contractile Dysfunction in Diabetics. Front Physiol 7:93

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