Naohiko Nakanishi

PARM-1 Is an Endoplasmic Reticulum Molecule Involved in Endoplasmic Reticulum Stress-Induced Apoptosis in Rat Cardiac Myocytes

To identify novel transmembrane and secretory molecules expressed in cardiac myocytes, signal sequence trap screening was performed in rat neonatal cardiac myocytes. One of the molecules identified was a transmembrane protein, prostatic androgen repressed message-1 (PARM-1). While PARM-1 has been identified as a gene induced in prostate in response to castration, its function is largely unknown. Our expression analysis revealed that PARM-1 was specifically expressed in hearts and skeletal muscles, and in the heart, cardiac myocytes, but not non-myocytes expressed PARM-1. Immunofluorescent staining showed that PARM-1 was predominantly localized in endoplasmic reticulum (ER). In Dahl salt-sensitive rats, high-salt diet resulted in hypertension, cardiac hypertrophy and subsequent heart failure, and significantly stimulated PARM-1 expression in the hearts, with a concomitant increase in ER stress markers such as GRP78 and CHOP. In cultured cardiac myocytes, PARM-1 expression was stimulated by proinflammatory cytokines, but not by hypertrophic stimuli. A marked increase in PARM-1 expression was observed in response to ER stress inducers such as thapsigargin and tunicamycin, which also induced apoptotic cell death. Silencing PARM-1 expression by siRNAs enhanced apoptotic response in cardiac myocytes to ER stresses. PARM-1 silencing also repressed expression of PERK and ATF6, and augmented expression of CHOP without affecting IRE-1 expression and JNK and Caspase-12 activation. Thus, PARM-1 expression is induced by ER stress, which plays a protective role in cardiac myocytes through regulating PERK, ATF6 and CHOP expression. These results suggested that PARM-1 is a novel ER transmembrane molecule involved in cardiac remodeling in hypertensive heart disease.

Circadian, weekly, and seasonal mortality variations in out-of-hospital cardiac arrest in Japan: analysis from AMI-Kyoto Multicenter Risk Study database.

BACKGROUND Several studies have reported circadian, weekly, and seasonal variations in the rates of out-of-hospital cardiac arrest (OHCA). However, variations in the mortality of OHCA are not well known. METHODS AND RESULTS We investigated the 1396 consecutive cases of OHCA with cardiac etiology between October 2004 and September 2008. There were 2 peaks in the occurrence of OHCA in early morning and late evening. There was a weekly pattern with an increased incidence on Mondays. We found a significant seasonal variation in the frequency of events, with a maximum during winter. There was a trend of reduced mortality in warmest 3 months, especially among a subgroup of ventricular fibrillation/pulseless ventricular tachycardia with arrest witnessed. CONCLUSION The present analyses demonstrated circadian, weekly and seasonal variations in the occurrence, and a seasonal variation in mortality in OHCA. Changes in temperature might influence the severity of OHCA and change the rate of success of cardiopulmonary resuscitation.

MURC/Cavin-4 facilitates recruitment of ERK to caveolae and concentric cardiac hypertrophy induced by α1-adrenergic receptors

Significance Caveolae are recognized as a platform for preassembled complexes of receptors, signal components, and their targets, facilitating efficient and specific cellular responses at the plasma membrane. ERK is activated at the plasma membrane and an important molecule that has been well studied for its integral role in signal transduction events during physiological adaptation and pathological manifestation. Here we show that although muscle-restricted coiled-coil protein (MURC)/Cavin-4, a muscle-specific caveola component, is dispensable for caveolar formation in cardiomyocytes, MURC/Cavin-4 serves as an ERK-recruiting protein in the caveolae within cardiomyocytes. The recruiting function of MURC/Cavin-4 is necessary to elicit efficient signaling of the α1-adrenergic receptor–ERK cascade in concentric cardiac hypertrophy. Our findings provide unique insight into the molecular mechanisms underlying caveola-mediated signaling in cardiac hypertrophy. The actions of catecholamines on adrenergic receptors (ARs) induce sympathetic responses, and sustained activation of the sympathetic nervous system results in disrupted circulatory homeostasis. In cardiomyocytes, α1-ARs localize to flask-shaped membrane microdomains known as “caveolae.” Caveolae require both caveolin and cavin proteins for their biogenesis and function. However, the functional roles and molecular interactions of caveolar components in cardiomyocytes are poorly understood. Here, we showed that muscle-restricted coiled-coil protein (MURC)/Cavin-4 regulated α1-AR–induced cardiomyocyte hypertrophy through enhancement of ERK1/2 activation in caveolae. MURC/Cavin-4 was expressed in the caveolae and T tubules of cardiomyocytes. MURC/Cavin-4 overexpression distended the caveolae, whereas MURC/Cavin-4 was not essential for their formation. MURC/Cavin-4 deficiency attenuated cardiac hypertrophy induced by α1-AR stimulation in the presence of caveolae. Interestingly, MURC/Cavin-4 bound to α1A- and α1B-ARs as well as ERK1/2 in caveolae, and spatiotemporally modulated MEK/ERK signaling in response to α1-AR stimulation. Thus, MURC/Cavin-4 facilitates ERK1/2 recruitment to caveolae and efficient α1-AR signaling mediated by caveolae in cardiomyocytes, which provides a unique insight into the molecular mechanisms underlying caveola-mediated signaling in cardiac hypertrophy.

The Increased Mortality from Witnessed Out-of-Hospital Cardiac Arrest in the Home

Abstract Background. Research in 2008 demonstrated that the majority of out-of-hospital cardiac arrests (OHCAs) occur in the home, and many important characteristics differ between private and public locations. However, the influence of the location of collapse on survival from OHCA is not well understood. Furthermore, most of the reports have been from Western countries; there is little research from Asia that differentiates the conditions of OHCA. Objective. To investigate the influence of the location of collapse on being discharged alive from OHCA and whether the location of collapse is also an independent predictor of survival from OHCA in Japan. Methods. We analyzed 463 consecutive cases of witnessed OHCA with cardiac etiology that occurred between October 2004 and September 2008 in Japan. We investigated the characteristics of OHCA patients who collapsed in private and public locations, and assessed the influence of the location of collapse on survival from OHCA. Results. Patients who collapsed outside the home were younger, more likely to be male, more likely to receive bystander cardiopulmonary resuscitation (CPR), and more likely to have ventricular fibrillation (VF)/pulseless ventricular tachycardia (VT) and had a shorter time interval between collapse and 9-1-1 call than patients who collapsed in the home. Mortality was significantly higher in the group who collapsed in the home. The independent influence of the location of collapse was eliminated by additional adjustment for time interval from collapse to 9-1-1 call, age, bystander CPR, and initial cardiac rhythm. Finally, VF/pulseless VT as the initial rhythm and bystander CPR were independently associated with the patients being discharged alive; the location of collapse was not an independently associated variable. Conclusions. The present analysis demonstrated that there were significant differences in survival between groups of patients who suffered from cardiac arrest inside and outside the home in Japan. The outside-the-home group had a higher rate of survival from OHCA; however, the location of collapse was not an independent predictor of survival from OHCA. Education of the families of high-risk patients in placing a rapid emergency call and performing effective CPR might be needed to improve survival from cardiac arrest in the home.

Two Cases of Delayed Cardiac Tamponade due to Pericarditis after Pulmonary Vein (PV) Isolation for Atrial Fibrillation

Catheter ablation is an established treatment for atrial fibrillation (AF). The incidence of major complications related to the procedure is reported to be 4.5%, and delayed cardiac tamponade (DCT) is a rare, although recently recognized, complication. However, the mechanisms underlying the development of DCT remain unclear. We herein report the cases of two men, both 49 years of age, who developed cardiac tamponade requiring pericardiocentesis a few weeks after undergoing pulmonary vein isolation for persistent AF. Physicians should explain to the patient the potential for DCT as a complication prior to performing catheter ablation and provide careful follow-up for at least a few weeks after the session.

Serglycin is a novel adipocytokine highly expressed in epicardial adipose tissue.

Much recent work has highlighted the key role of adipose tissue as an endocrine organ that secretes a number of adipocytokines, linking adiposity, especially intra-abdominal visceral fat, and the pathogenesis of cardiovascular and metabolic diseases. However, the role of epicardial adipose tissue (EAT), another important visceral fat depot situated in close proximity to epicardial coronary arteries and myocardium, has been less well studied. In this study, we sought to characterize EAT by comparing gene expression profiles of EAT, omental adipose tissue (OAT), and subcutaneous adipose tissue (SCAT) in patients who underwent elective coronary artery bypass graft surgery for critical coronary artery disease (CAD) and identify molecules involved in inflammation. A total of 15,304 probes were detected in all depots, and 231 probes were differentially expressed. Significantly higher expression of pro-inflammatory genes such as interleukin-1β, -6, and -8, and chemokine receptor 2 was observed in EAT, even when compared with OAT. Among them, serglycin was one of the most abundantly expressed genes in EAT. Serglycin expression was induced during adipocytic differentiation of 3T3L1 cells. Serglycin was secreted from adipocytes, and tumor necrosis factor-α stimulated its expression and secretion in adipocytes. Serglycin was also present in human serum samples. These results suggest that human EAT has strong inflammatory properties in patients with CAD and provide novel evidence that serglycin is an adipocytokine highly expressed in EAT.

The coiled-coil domain of MURC/cavin-4 is involved in membrane trafficking of caveolin-3 in cardiomyocytes

Muscle-restricted coiled-coil protein (MURC), also referred to as cavin-4, is a member of the cavin family that works cooperatively with caveolins in caveola formation and function. Cavins are cytoplasmic proteins with coiled-coil domains and form heteromeric complexes, which are recruited to caveolae in cells expressing caveolins. Among caveolins, caveolin-3 (Cav3) is exclusively expressed in muscle cells, similar to MURC/cavin-4. In the heart, Cav3 overexpression contributes to cardiac protection, and its deficiency leads to progressive cardiomyopathy. Mutations in the MURC/cavin-4 gene have been identified in patients with dilated cardiomyopathy. In the present study, we show the role of MURC/cavin-4 as a caveolar component in the heart. In H9c2 cells, MURC/cavin-4 was localized at the plasma membrane, whereas a MURC/cavin-4 mutant lacking the coiled-coil domain (ΔCC) was primarily localized to the cytoplasm. ΔCC bound to Cav3 and impaired membrane localization of Cav3 in cardiomyocytes. Additionally, although ΔCC did not alter Cav3 mRNA expression, ΔCC decreased the Cav3 protein level. MURC/cavin-4 and ΔCC similarly induced cardiomyocyte hypertrophy; however, ΔCC showed higher hypertrophy-related fetal gene expression than MURC/cavin-4. ΔCC induced ERK activation in cardiomyocytes. Transgenic mice expressing ΔCC in the heart (ΔCC-Tg mice) showed impaired cardiac function accompanied by cardiomyocyte hypertrophy and marked interstitial fibrosis. Hearts from ΔCC-Tg mice showed a reduction of the Cav3 protein level and activation of ERK. These results suggest that MURC/cavin-4 requires its coiled-coil domain to target the plasma membrane and to stabilize Cav3 at the plasma membrane of cardiomyocytes and that MURC/cavin-4 functions as a crucial caveolar component to regulate cardiac function.

Angioscopic observation in chronic thromboembolic pulmonary hypertension before and after balloon pulmonary angioplasty

: We here report the angioscopic assessment of inoperable peripheral chronic thromboembolic pulmonary hypertension in an 81-year-old man who was previously diagnosed with the disease. To assess the pathological morphology of the web lesion, pouch defect and band lesion, we used two types of angioscopic catheter: blood flow-maintaining type and blood flow-blocking type. Angioscopy revealed a heterogeneous thrombus that contained white organized thrombus, red fragile thrombus and yellowish thrombus. After three sessions of balloon pulmonary angioplasty, his pulmonary arterial pressure decreased and his symptom of dyspnea on exertion and desaturation were improved. Angioscopy displayed various forms of organized thrombus, and also allowed the detailed observation of lesions that were difficult to be observed by angiography.

PTRF/Cavin-1 Deficiency Causes Cardiac Dysfunction Accompanied by Cardiomyocyte Hypertrophy and Cardiac Fibrosis

Mutations in the PTRF/Cavin-1 gene cause congenital generalized lipodystrophy type 4 (CGL4) associated with myopathy. Additionally, long-QT syndrome and fatal cardiac arrhythmia are observed in patients with CGL4 who have homozygous PTRF/Cavin-1 mutations. PTRF/Cavin-1 deficiency shows reductions of caveolae and caveolin-3 (Cav3) protein expression in skeletal muscle, and Cav3 deficiency in the heart causes cardiac hypertrophy with loss of caveolae. However, it remains unknown how loss of PTRF/Cavin-1 affects cardiac morphology and function. Here, we present a characterization of the hearts of PTRF/Cavin-1-null (PTRF−/−) mice. Electron microscopy revealed the reduction of caveolae in cardiomyocytes of PTRF−/− mice. PTRF−/− mice at 16 weeks of age developed a progressive cardiomyopathic phenotype with wall thickening of left ventricles and reduced fractional shortening evaluated by echocardiography. Electrocardiography revealed that PTRF−/− mice at 24 weeks of age had low voltages and wide QRS complexes in limb leads. Histological analysis showed cardiomyocyte hypertrophy accompanied by progressive interstitial/perivascular fibrosis. Hypertrophy-related fetal gene expression was also induced in PTRF−/− hearts. Western blotting analysis and quantitative RT-PCR revealed that Cav3 expression was suppressed in PTRF−/− hearts compared with that in wild-type (WT) ones. ERK1/2 was activated in PTRF−/− hearts compared with that in WT ones. These results suggest that loss of PTRF/Cavin-1 protein expression is sufficient to induce a molecular program leading to cardiomyocyte hypertrophy and cardiomyopathy, which is partly attributable to Cav3 reduction in the heart.

MURC deficiency in smooth muscle attenuates pulmonary hypertension

Emerging evidence suggests that caveolin-1 (Cav1) is associated with pulmonary arterial hypertension. MURC (also called Cavin-4) is a member of the cavin family, which regulates caveolar formation and functions together with caveolins. Here, we show that hypoxia increased Murc mRNA expression in the mouse lung, and that Murc-null mice exhibited attenuation of hypoxia-induced pulmonary hypertension (PH) accompanied by reduced ROCK activity in the lung. Conditional knockout mice lacking Murc in smooth muscle also resist hypoxia-induced PH. MURC regulates the proliferation and migration of pulmonary artery smooth muscle cells (PASMCs) through Rho/ROCK signalling. Cav1 suppresses RhoA activity in PASMCs, which is reversed by MURC. MURC binds to Cav1 and inhibits the association of Cav1 with the active form of Gα13, resulting in the facilitated association of the active form of Gα13 with p115RhoGEF. These results reveal that MURC has a function in the development of PH through modulating Rho/ROCK signalling.