Reactive oxygen species (ROS) play both physiological and pathological roles in the heart and critically regulate survival and death of cardiomyocytes (CMs). NADPH oxidases purposefully produce ROS, including superoxide and hydrogen peroxide, by transferring electrons from NADPH or NADH to molecular oxygen. We have been focusing on NADPH oxidase 4 (Nox4), one of the major NADPH oxidase isoforms expressed in intracellular membranes in the heart, and we have discovered its key features. Excessive activation of Nox4 in response to hypertrophic stimuli, aging, and ischemia/reperfusion (I/R) leads to increases in oxidative stress, mitochondrial dysfunction, CM cell death, and cardiac dysfunction. On the other hand, basal production of ROS by either Nox2 or Nox4 during ischemia and reperfusion acts as a physiological second messenger to induce hypoxic adaptation. These studies suggest that the activity of Nox4 is supposed to be kept at appropriate levels in order to avoid accumulation of ROS either above or below physiological levels. How the activity of Nox4 is controlled in CMs under various pathophysiological conditions is poorly understood. Nox4 is believed to be regulated primarily at the level of transcription/translation. However, using yeast two-hybrid screening, we have identified that Nox4 physically associates with Fyn, a non-receptor type tyrosine kinase and that Fyn- mediated phosphorylation of Nox4 negatively regulates Nox4 and serves as an important negative feedback mechanism to limit excessive oxidative stress in the heart. We here test our hypotheses 1) that Fyn-mediated phosphorylation of Nox4 at Y566 inhibits interaction between Nox4 and p22phox and negatively regulates ROS production by Nox4, and 2) that activation of endogenous Fyn in response to PO and ischemic preconditioning (IPC) induces phosphorylation/suppression of Nox4 at Y566, thereby attenuating the NADPH oxidase activity of Nox4 and protecting the heart from PO and I/R injury. Using both molecular signaling and in vivo physiology approaches, we will show the molecular mechanism of the Fyn-mediated suppression of Nox4 and the functional significance of Fyn-mediated regulation of Nox4 during PO and IPC in vivo. Our study will elucidate a novel and direct PTM mechanism by which Nox4, the major ROS-producing enzyme in the heart, is regulated by stress and its functional significance during cardiac conditions frequesntly associated with aging. Our study will provide an important clue to better control the activity of Nox4 in the heart during cardiac stress in cardiovascular patients.

Public Health Relevance

Excessive production of free radicals and peroxide, termed reactive oxygen species (ROS), in response to stresses, including high blood pressure and heart attack, causes malfunction and death of heart cells and leads to heart failure in elderly patients. We will investigate how the function of an enzyme termed Nox4, an important source of ROS in the heart, is modulated and how this mechanism affects cardiac damage during cardiac stress. Knowledge obtained from this project should lead to a novel strategy to modify the activity of Nox4, thereby preventing cardiac damage caused by excessive ROS in patients.

National Institute of Health (NIH)
Research Project (R01)
Project #
Application #
Study Section
Cardiac Contractility, Hypertrophy, and Failure Study Section (CCHF)
Program Officer
Kohanski, Ronald A
Project Start
Project End
Budget Start
Budget End
Support Year
Fiscal Year
Total Cost
Indirect Cost
Rutgers University
Anatomy/Cell Biology
Schools of Medicine
United States
Zip Code
Maejima, Yasuhiro; Chen, Yun; Isobe, Mitsuaki et al. (2015) Recent progress in research on molecular mechanisms of autophagy in the heart. Am J Physiol Heart Circ Physiol 308:H259-68
Sciarretta, Sebastiano; Volpe, Massimo; Sadoshima, Junichi (2014) NOX4 regulates autophagy during energy deprivation. Autophagy 10:699-701
Del Re, Dominic P; Sadoshima, Junichi (2014) Elucidating ERK2 function in the heart. J Mol Cell Cardiol 72:336-8
Shao, Dan; Zhai, Peiyong; Del Re, Dominic P et al. (2014) A functional interaction between Hippo-YAP signalling and FoxO1 mediates the oxidative stress response. Nat Commun 5:3315
Wu, Changgong; Jain, Mohit Raja; Li, Qing et al. (2014) Identification of novel nuclear targets of human thioredoxin 1. Mol Cell Proteomics 13:3507-18
Ikeda, Yoshiyuki; Sciarretta, Sebastiano; Nagarajan, Narayani et al. (2014) New insights into the role of mitochondrial dynamics and autophagy during oxidative stress and aging in the heart. Oxid Med Cell Longev 2014:210934
Matsushima, Shouji; Tsutsui, Hiroyuki; Sadoshima, Junichi (2014) Physiological and pathological functions of NADPH oxidases during myocardial ischemia-reperfusion. Trends Cardiovasc Med 24:202-5
Yamamoto, Takanobu; Byun, Jaemin; Zhai, Peiyong et al. (2014) Nicotinamide mononucleotide, an intermediate of NAD+ synthesis, protects the heart from ischemia and reperfusion. PLoS One 9:e98972
Maejima, Yasuhiro; Sadoshima, Junichi (2014) SUMOylation: a novel protein quality control modifier in the heart. Circ Res 115:686-9
Maejima, Yasuhiro; Usui, Soichiro; Zhai, Peiyong et al. (2014) Muscle-specific RING finger 1 negatively regulates pathological cardiac hypertrophy through downregulation of calcineurin A. Circ Heart Fail 7:479-90

Showing the most recent 10 out of 69 publications