Rho-associated kinases (ROCKs), the immediate downstream targets of RhoA, are ubiquitously expressed serine-threonine protein kinases that are involved in diverse cellular functions, including smooth muscle contraction, actin cytoskeleton organization, cell adhesion and motility, and gene expression. Recent studies have shown that ROCKs may play a pivotal role in cardiovascular diseases such as vasospastic angina, ischemic stroke, and heart failure. Indeed, inhibition of ROCKs by statins or other selective inhibitors leads to the upregulation and activation of endothelial nitric oxide synthase (eNOS) and reduction of vascular inflammation and atherosclerosis. Thus inhibition of ROCKs may contribute to some of the cholesterol-independent beneficial effects of statin therapy. Currently, two ROCK isoforms have been identified, ROCK1 and ROCK2. Because ROCK inhibitors are nonselective with respect to ROCK1 and ROCK2 and also, in some cases, may be nonspecific with respect to other ROCK-related kinases such as myristolated alanine-rich C kinase substrate (MARCKS), protein kinase A, and protein kinase C, the precise role of ROCKs in cardiovascular disease remains unknown. However, with the recent development of ROCK1- and ROCK2-knockout mice, further dissection of ROCK signaling pathways is now possible. Herein we review what is known about the physiological role of ROCKs in the cardiovascular system and speculate about how inhibition of ROCKs could provide cardiovascular benefits.

Background-Rho GTPase and its downstream target, Rho-associated kinase (ROCK), have been implicated in diverse cardiovascular diseases such as cardiac hypertrophy. However, pharmacological inhibitors of ROCK are not entirely specific, nor can they discriminate between the ROCK isoforms ROCK1 and ROCK2. To determine the specific role of ROCK1 in the development of cardiac hypertrophy, we generated ROCK1 ϩ/Ϫ haploinsufficient mice and determined whether cardiac hypertrophy and remodeling are decreased in these mice. Methods and Results-Litters of ROCK1Ϫ/Ϫ mice on C57Bl/6 background were markedly underrepresented, suggesting lethality in utero or postnatally. ROCK1 ϩ/Ϫ mice, however, are viable and fertile with no obvious phenotypic abnormalities. Basal blood pressure, heart rate, and cardiac dimension and function in ROCK1 ϩ/Ϫ mice were similar to those in wild-type (WT) littermates. Infusion of angiotensin II (400 ng · kg Ϫ1 · min Ϫ1 for 28 days) or treatment with N G -nitro-L-arginine methyl ester (1 mg/mL in drinking water for 28 days) caused similar increases in systolic blood pressure, left ventricular wall thickness, left ventricular mass, ratio of heart weight to tibial length, and cardiomyocyte size in ROCK1 ϩ/Ϫ mice and WT littermates. In contrast, perivascular fibrosis in hearts was increased to a lesser extent in ROCK1ϩ/Ϫ mice compared with WT littermates. This was associated with decreased expression of transforming growth factor-␤, connective tissue growth factor, and type III collagen. In addition, perivascular fibrosis induced by transaortic constriction or myocardial infarction was decreased in ROCK1 ϩ/Ϫ mice compared with WT littermates. Conclusions-These findings indicate ROCK1 is critical for the development of cardiac fibrosis, but not hypertrophy, in response to various pathological conditions and suggest that signaling pathways leading to the hypertrophic and profibrotic response of the heart are distinct.

Although Rho-associated kinase (ROCK) activity has been implicated in cardiovascular diseases, the tissueand isoform-specific roles of ROCKs in the vascular response to injury are not known. To address the role of ROCKs in this process, we generated haploinsufficient Rock1 (Rock1 +/-) and Rock2 (Rock2 +/-) mice and performed carotid artery ligations. Following this intervention, we found reduced neointima formation in Rock1 +/-mice compared with that of WT or Rock2 +/-mice. This correlated with decreased vascular smooth muscle cell proliferation and survival, decreased levels proinflammatory adhesion molecule expression, and reduced leukocyte infiltration. In addition, thioglycollate-induced peritoneal leukocyte recruitment and accumulation were substantially reduced in Rock1 +/-mice compared with those of WT and Rock2 +/-mice. To determine the role of leukocyte-derived ROCK1 in neointima formation, we performed reciprocal bone marrow transplantation (BMT) in WT and Rock1 +/-mice. Rock1 +/-to WT BMT led to reduced neointima formation and leukocyte infiltration following carotid ligation compared with those of WT to WT BMT. In contrast, WT to Rock1 +/-BMT resulted in increased neointima formation. These findings indicate that ROCK1 in BM-derived cells mediates neointima formation following vascular injury and suggest that ROCK1 may represent a promising therapeutic target in vascular inflammatory diseases.

Background Notch1 regulates binary cell fate determination and is critical for angiogenesis and cardiovascular development. However, the pathophysiological role of Notch1 in the postnatal period is not known. We hypothesize that Notch1 signaling in vascular smooth muscle cells (SMC) may contribute to neointimal formation following vascular injury. Methods and Results We performed carotid artery ligation in wild-type (WT), control (smCre-Tg), general Notch1 heterozygous deficient (N1+/-), SMC-specific Notch1 heterozygous deficient (smN1+/-), and general Notch3 homozygous deficient (N3-/-) mice. Compared to WT or control mice, N1+/- and smN1+/- mice showed a 70% decrease in neointimal formation following carotid artery ligation. However, neointimal formation was similar between WT and N3-/- mice. Indeed, SMC derived from explanted aortas of either N1+/- or smN1+/- mice showed decreased chemotaxis and proliferation, and increased apoptosis compared to control or N3-/- mice. This correlated with decreased staining of PCNA positive cells and increased staining of cleaved Caspase-3 in the intima of N1+/- or smN1+/- mice. In SMC derived from CHF1/Hey2-/- mice, activation of Notch signaling did not lead to increase SMC proliferation or migration. Conclusion These findings indicate that Notch1, rather than Notch3, mediates SMC proliferation and neointimal formation following vascular injury through CHF1/Hey2 and suggest that therapies, which target Notch1/CHF1/Hey2 in SMC, may be beneficial in preventing vascular proliferative diseases.

Rho kinases (ROCKs) are serine-threonine protein kinases that regulate the actin cytoskeleton. Recent studies suggest that ROCKs also play an important role in cardiovascular disease. However, the isoform- and tissue-specific role of ROCKs in mediating this process is unknown. Using homologous recombination, we generated mutant mice harboring alleles with homozygous deletion of ROCK1 (ROCK1(-/-)). Most ROCK1(-/-) mice die perinatally. However, a few ROCK1(-/-) mice survive to adulthood, are phenotypically normal, and have no apparent compensatory changes in ROCK2. Using these ROCK1(-/-) mice, we show that ROCK1 in bone marrow-derived macrophages is critical to the development of atherosclerosis, in part, by mediating foam cell formation and macrophage chemotaxis. Lipid accumulation and atherosclerotic lesions were reduced in atherosclerosis-prone LDLR(-/-) mice, whose bone marrows have been replaced with bone marrows derived from ROCK1(-/-) mice. Bone marrow-derived ROCK1-deficient macrophages exhibited impaired chemotaxis to monocyte chemotactic protein-1 and showed reduced ability to take up lipids and to develop into foam cells when exposed to modified low-density lipoprotein. These findings indicate that ROCK1 in bone marrow-derived cells is a critical mediator of atherogenesis and suggest that macrophage ROCK1 may be an important therapeutic target for vascular inflammation and atherosclerosis.

SUMMARYA 37 year-old female who had suffered from arteritis for 20 years underwent a Bentall operation. Since severe stenosis was observed in her left main coronary artery (LMCA) the following year, a minimally invasive direct coronary artery bypass (MIDCAB) operation was performed. Unfortunately, she again complained of angina about 6 months after the second surgery and coronary angiography (CAG) revealed that her left internal thoracic artery graft was totally occluded. Although a 4.0 × 15 mm S670 stent was placed in her LMCA, the LMCA restenosed every 3 months and she underwent reintervention 8 times. We placed 2 sirolimus-eluting stents for treating the LMCA using the culottes stenting technique. CAG 6 months after the index procedure showed no stenosis at her LMCA. Sirolimus-eluting stents were effective for treating stenosis resulting from arteritis as well as that caused by atherosclerosis. ( SIROLIMUS has an antiproliferative effect against vascular smooth muscle cells, and many investigators have already reported that sirolimus-eluting stent (SES) placement is more effective for treating atherosclerotic coronary artery narrowing than conventional bare metal stent placement. However, there have been very few reports on whether SES placement is effective against coronary artery stenosis caused by arteritis.We report herein the case of a young female patient suffering from arteritis in whom malignant instent restenosis was treated successfully with an SES. This is a rare case showing the efficacy of SES for treating LMCA stenosis caused by arteritis.From the

Background: Vildagliptin is a dipeptidyl peptidase-4 inhibitor that reduces glycemia in patients with type 2 diabetes mellitus (T2DM). When approved in 2013, data on vildagliptin combined with >750 mg/ day metformin in Japanese patients were limited. There is a need to confirm the safety and efficacy of vildagliptin in combination with oral antidiabetic drugs (OADs). Research design and methods: This 52-week post-marketing surveillance (PMS) observational study in Japanese T2DM patients evaluated the safety and efficacy of vildagliptin in combination with OADs including high-dose metformin or insulin but excluding combination with sulfonylureas alone. Results: During this survey of 3006 Japanese T2DM patients, 13.61% of patients experienced adverse events (AEs) and 2.20% reported a serious AE (SAE). The frequency of AEs/SAEs was similar when in combination with biguanides (12.93%/1.46%), metformin ≥1000 mg/day (12.92%/1.22%), metformin <1000 mg/day (12.62%/1.54%), thiazolidine derivatives (16.71%/2.86%), α-glucosidase inhibitors (13.18%/1.90%), rapid-acting insulin secretagogues (glinides) (20.41%/5.71%), or insulin (15.87%/ 2.47%). The mean ± SD changes from baseline at endpoint in glycated hemoglobin and fasting blood glucose were −0.76 ± 1.27% and −23.3 ± 57.3 mg/dL, respectively, and these changes were consistent, regardless of concomitant OAD. Conclusions: Long-term vildagliptin combination therapy is safe and effective in Japanese T2DM patients in real-world settings.

Background: Our objective was to characterize cases of hospitalized heart failure (HHF) focusing on inhospital resource utilization (particularly furosemide doses) and worsening heart failure (WHF), and identify which factors are associated with the length of stay (LOS). Methods: Cases of HHF (!20 years), excluding those undergoing surgical procedures and in-hospital deaths, were retrieved from the Japanese Diagnosis Procedure Combination database (April 2012 to March 2016). WHF was defined using eight components, including up-titration of intravenous drugs and non-pharmacological management. Results: The mean age of 78,953 cases of HHF was 79 years and 51% were male. The median LOS was 17 days. The maximum daily dose and cumulative dose of furosemide (mean AE standard deviation) were 43.3 AE 56.0 mg and 215.6 þ 450.6 mg, respectively, for intravenous furosemide, and 44.0 AE 37.3 mg and 523.3 AE 675.4 mg, respectively, for oral furosemide. The incidence of WHF was 36.1% during hospitalization and 19.3% from 6th hospital day to discharge. The mean number of WHF components was 1.4 AE 0.7 during hospitalization and 1.3 AE 0.6 from 6th hospital day. Regression analyses showed that the number of WHF components from 6th hospital day, pneumonia, and hyponatremia were strongly associated with longer LOS. Conclusions: These findings in patients with HHF could be vital to focus future efforts to improve the therapeutic strategies for heart failure.