H. Kawajiri

Inhibitory effect of a TGFβ receptor type-I inhibitor, Ki26894, on invasiveness of scirrhous gastric cancer cells

Background:Gastric cancer cells frequently metastasise, partly because of their highly invasive nature. Transforming growth factor-β (TGF-β) receptor signalling is closely associated with the invasion of cancer cells. The aim of this study was to clarify the effect of a TGF-β receptor (TβR) phosphorylation inhibitor on the invasiveness of gastric cancer cells.Methods:Four gastric cancer cell lines, including two scirrhous-type cell lines and two non-scirrhous-type cell lines, were used. A TβR type I (TβR-I) kinase inhibitor, Ki26894, inhibits the phosphorylation of Smad2 at an ATP-binding site of TβR-I. We investigated the expression levels of TβR and phospho-Smad2, and the effects of TGF-β in the presence or absence of Ki26894 on Smad2 phosphorylation, invasion, migration, epithelial-to-mesenchymal transition (EMT), Ras homologue gene family member A (RhoA), ZO-2, myosin, and E-cadherin expression of gastric cancer cells.Results:TβR-I, TβR-II, and phospho-Smad2 expressions were found in scirrhous gastric cancer cells, but not in non-scirrhous gastric cancer cells. Ki26894 decreased Smad2 phosphorylation induced by TGF-β1 in scirrhous gastric cancer cells. Transforming growth factor-β1 upregulated the invasion, migration, and EMT ability of scirrhous gastric cancer cells. Transforming growth factor-β1 significantly upregulated the activity of RhoA and myosin phosphorylation, whereas TGF-β1 decreased ZO-2 and E-cadherin expression in scirrhous gastric cancer cells. Interestingly, Ki26894 inhibited these characteristics in scirrhous gastric cancer cells. In contrast, non-scirrhous gastric cancer cells were not affected by TGF-β1 or Ki26894 treatment.Conclusion:A TβR-I kinase inhibitor decreases the invasiveness and EMT of scirrhous gastric cancer cells. Ki26894 is therefore considered to be a promising therapeutic compound for the metastasis of scirrhous gastric carcinoma.

Significance of E-cadherin expression in triple-negative breast cancer

Purpose:Triple-negative breast cancer (TNBC), a subtype of breast cancer that is oestrogen receptor (ER) negative, progesterone receptor (PR) negative, and human epidermal growth factor receptor 2 (HER2) negative, has a poor prognosis. Although a correlation between E-cadherin expression level and outcome has been demonstrated among all types of breast cancer, little is known about the significance of E-cadherin expression levels in TNBC.Methods:A total of 574 patients who had undergone a resection of a primary breast cancer except for invasive lobular carcinomas were enrolled in this study. Expressions of ER, PR, HER2, and E-cadherin were assessed by immunohistochemistry. We examined the association between TNBC and other clinicopathological variables and evaluated the significance of the E-cadherin expression.Results:Among the 574 breast cancer cases, 123 (21.4%) revealed a triple-negative phenotype. Patients with TNBC experienced more frequent lymph node metastasis (P=0.024) and a poorer prognosis (P<0.001) in comparison with non-TNBC patients. Triple-negative breast cancer was an independent prognostic factor. Reduced levels of E-cadherin were observed in 238 (41.5%) of the 574 breast cancer cases. E-cadherin reduction was significantly frequent in cases of TNBC (P<0.001) and lymph node metastasis (P=0.032). Furthermore, in the 123 TNBC cases, the prognosis of patients with an E-cadherin-negative expression was significantly worse than that of E-cadherin-positive patients (P=0.0265), especially for those in clinical stage II (P=0.002). A multivariate logistic regression analysis showed a reduction of the E-cadherin expression to be an independent prognostic factor (P=0.046).Conclusion:E-cadherin expression may be a useful prognostic marker for classifying subgroups of TNBC.

c‐Kit expression as a prognostic molecular marker in patients with basal‐like breast cancer

As patients with basal‐like breast cancer (BLBC) have a poor prognosis and there is no specifically tailored therapy, molecular biological characterization of BLBC is necessary. c‐Kit is a transmembrane receptor tyrosine kinase known to play important roles in various solid cancers. This study classified BLBCs from patients with breast carcinoma, and addressed the significance of c‐Kit expression in these tumours.

Predictive impact of daily physical activity on new vascular events in patients with mild ischemic stroke.

Background Daily physical inactivity is associated with a substantially increased risk of cardiovascular events. However, the target level of daily physical activity remains unclear. Aim We aimed to evaluate the impact of physical activity on long-term vascular events in patients with mild ischemic stroke. Methods We designed a single hospital-based prospective observational study and studied 166 ischemic stroke patients (mean age: 63·9 ± 9·2) who had a modified Rankin Scale 0–1. We measured the daily step count as a variable of the daily physical activity after three-months from the stroke onset. Other clinical characteristics including age, body mass index, blood pressure, blood laboratory tests, vascular function and medications were also assessed. The primary outcomes were hospitalization due to stroke recurrence, myocardial infarction, angina pectoris and peripheral artery disease. Survival curves were calculated by a Kaplan–Meier survival analysis, and the hazard ratios for recurrences were determined by univariate and multivariate Cox proportional hazards regression models. Results After a median follow-up periods of 1332 days, 34 vascular events (23 stroke recurrences, 11 coronary artery disease) and 7 drop-outs occurred, and the remaining patients were divided into two groups: the without recurrence group (n = 125) and the with recurrence group (n = 34). The daily step count was lower in the nonsurvivor group than in the survivor group. Univariate and multivariate Cox proportional hazards analyses revealed that the daily step counts was independent predictors of new vascular events. A daily step count cutoff value of 6025 steps per day was determined by analyzing the receiver-operating characteristics that showed a sensitivity of 69·4% and a specificity of 79·4%. The Kaplan–Meier survival curves after a log-rank test showed a significantly lower event rate in the more than 6025 steps per day group compared with the less than 6025 steps per day group (P = 0·0002). The positive and negative predictive values of less than 6025 steps were 38·0% and 91·6%, respectively. Conclusion Our data indicate that daily physical activity evaluated by step counts may be useful for forecasting the prognosis in patients with mild ischemic stroke. Daily step counts of approximately 6000 steps per day may be an initial target level for reducing new vascular events.

High-risk cardiac surgery as an alternative to transplant or mechanical support in patients with end-stage heart failure

Objective Although the results of cardiac surgery in patients with poor left ventricular function have been widely published, the outcomes in patients with end‐stage heart failure who meet criteria for advanced therapies are not well investigated. As access to transplantation and ventricular assist device therapy remains limited, we explored the possibility of conventional surgery as an alternative option for highly selected patients with end‐stage heart failure. Methods We identified patients with left ventricular ejection fraction <20% and VO2 max <14 mL/min/m2, who were initially referred for advanced therapies but were instead offered a conventional procedure from 2002 to 2012. We examined the short‐ and midterm outcomes and compared survival with that after our advanced therapies in the same era. Results A total of 133 patients were identified; 68 were deemed to be transplant‐eligible, whereas 65 were transplant‐ineligible. Seventy‐nine percent were in New York Heart Association class III/IV. In‐hospital mortality was 12%. Actuarial survival at 5 and 10 years was 72% ± 4% and 39% ± 8%, respectively. Nonischemic etiology was identified as a predictor of late mortality. In the propensity‐adjusted model, our transplant‐eligible patients had comparable long‐term survival to our transplantation patients (HR 1.48 [95% confidence interval, 0.66‐3.2], P = .34), whereas the survival in our transplant‐ineligible subset was comparable to the survival after our left ventricular assist device therapy (HR 0.49 [95% confidence interval, 0.16‐1.50], P = .21). Conclusions Despite high perioperative risk, the midterm survival after conventional surgery in patients eligible for advanced therapies seems to be acceptable and may be an alternative option for highly selected patients with end‐stage heart failure.

Longitudinal Assessment of Inflammation in Recipients of Continuous-Flow Left Ventricular Assist Devices

BACKGROUND The long-term effects of continuous-flow left ventricular assist device (CF-LVAD) support on trends of inflammatory markers over time are unknown. We examined the hypothesis that the levels of inflammatory markers in CF-LVAD recipients are higher than in healthy controls and that these levels increase over time with long-term CF-LVAD support. METHODS We examined the levels of inflammatory markers longitudinally at baseline before CF-LVAD implantation and at 3, 6, and 9 months after implantation. We then compared the levels of inflammatory markers to those in a healthy control group. RESULTS Compared with baseline values before CF-LVAD implantation, left ventricular end-diastolic diameter (LVEDd) and left ventricular end-systolic diameter (LVESd) decreased significantly at 3, 6, and 9 months after CF-LVAD implantation. Brain natriuretic peptide (BNP) levels dropped significantly after CF-LVAD implantation but did not normalize. Improvements in ejection fraction at 3, 6, and 9 months after CF-LVAD implantation did not reach significance. Monocyte chemoattractant protein-1, interferon γ-induced protein, and C-reactive protein levels were higher in the CF-LVAD recipients at each of the time points (baseline before CF-LVAD implantation and 3, 6, and 9 months after implantation) compared with levels in healthy controls. In CF-LVAD recipients, serum interleukin-8, tumour necrosis factor-α, and macrophage inflammatory protein-β increased significantly at 9 months, and macrophage-derived chemokine increased at 6 months after CF-LVAD implantation compared with baseline. CONCLUSIONS Despite improvements in LV dimensions and BNP levels, markers of inflammation remained higher in CF-LVAD recipients. High levels of inflammation in CF-LVAD recipients may result from heart failure preconditioning or the long-term device support, or both. Because inflammation may be detrimental to CF-LVAD recipients, future studies should determine whether inflammatory pathways are reversible.

Tricuspid Valve Annular Dilation as a Predictor of Right Ventricular Failure After Implantation of a Left Ventricular Assist Device.

Tricuspid annular (TA) dilation has been suggested as a more reliable marker of concomitant advanced right ventricular failure (RVF) than severity of tricuspid regurgitation (TR). Our objective was to examine the impact of TA dilation on occurrence of RVF and in‐hospital mortality following left ventricular assist device (LVAD) implant.

173PTLE3 IS A USEFUL MARKER FOR PREDICTING THE THERAPEUTIC EFFECT OF ERIBULIN CHEMOTHERAPY FOR TRIPLE-NEGATIVE BREAST CANCER

ABSTRACT Aim: The recently developed reagent, eribulin mesylate (eribulin), is a microtubule dynamics inhibitor with a mechanism of action that differs from that of taxanes and vinca alkaloids. This drug is considered to be a promising chemotherapeutic agent for the treatment of locally advanced or metastatic breast cancer (MBC). In this study, we investigated predictive factors with respect to the therapeutic effect of eribulin chemotherapy among variables such as b-tubulin class III, GSTP1 and TLE3, which have previously been reported to be predictive factors of the therapeutic effect of taxanes, in an aim to identify possible biomarkers for predicting the efficacy of eribulin. Methods: The subjects included 52 patients with MBC who underwent chemotherapy using eribulin. The median follow-up time was 431 days (range, 50-650 days). The overall response rate (ORR), clinical benefit rate (CBR), disease control rate (DCR), overall survival (OS), time to treatment failure (TTF) and progression-free survival (PFS) were calculated regarding the efficacy of this regimen. Moreover, breast cancer was classified into intrinsic subtypes according to the status of the ER, PR, HER2 and Ki67 expression. Results: The clinical effects were as follows: overall ORR= 34.6%, CBR = 44.2%, DCR= 51.9%; median OS = 334 days; median TTF = 81 days; and median PFS = 275 days. TLE3, b-tubulin class III and GSTP1 were expressed in 24 cases, 21 cases and 24 cases, respectively, among the 52 patients investigated. The expression of TLE3 was found significantly more frequently in the TNBC lesions than in the non-TNBC lesions (p = 0.030). The patients with TLE3-negative tumors experienced significantly poorer outcomes in terms of progression-free survival when comparing the prognosis within the group of patients with triple-negative breast cancer (TNBC) lesions (p = 0.011). Based on a multivariate logistic regression analysis including 22 patients with TNBC also showed that a positive TLE3 expression significantly correlated with a better progression-free survival (p = 0.037). Conclusions: Our findings suggest that TLE3 is a useful marker for predicting the therapeutic effect of eribulin chemotherapy for TNBC. Disclosure: All authors have declared no conflicts of interest.

Maximum Walking Speed at Discharge Could Be a Prognostic Factor for Vascular Events in Patients With Mild Stroke: A Cohort Study

OBJECTIVE To identify the prognostic value of physical activity-related factors as well as known vascular risk factors for vascular events in mild ischemic stroke (MIS). DESIGN Single-center prospective cohort study. SETTING University hospital. PARTICIPANTS Consecutive patients (N=255) (175 men, median age 70.0y) with acute ischemic stroke and transient ischemic attack (TIA) with modified Rankin scale scores ranging from 0 to 2 were enrolled in this study. INTERVENTIONS Not applicable. MAIN OUTCOME MEASURES Enrolled patients were followed up for composite vascular events as primary outcomes up to 3 years postdischarge. Primary outcomes included stroke and cardiovascular death, hospitalization due to stroke or TIA recurrence, cardiovascular disease, and peripheral artery disease. During hospitalization, known vascular risk factors such as previous history of vascular events, stroke subtype, white matter lesions, and ankle-brachial index were assessed. Moreover, at the time of discharge, physical activity-related factors such as maximum walking speed (MWS), handgrip strength, knee extensor isometric muscle strength, anxiety, and depression were assessed as potential predictors. RESULTS The Kaplan-Meier estimates of cumulative risk of composite vascular events at 1, 2, and 3 years were 9.6%, 14.4%, and 15.2%, respectively. After multivariate analysis, cerebral white matter lesions of periventricular hyperintensity (PVH) (grade=3; hazard ratio: 2.904; 95% confidence interval: 1.160 to 7.266; P=.023) and MWS (<1.45m/s; hazard ratio: 2.232; 95% confidence interval: 1.010 to 4.933; P=.047) were identified as significant independent predictors of composite vascular events. CONCLUSIONS The results of this study indicate that MWS could be an independent prognostic factor for composite vascular events in MIS.

TERT-BUTYLHYDROQUINONE RESCUES CYCLOSPORINE-A MEDIATED IMPAIRMENT IN VASCULAR FUNCTION VIA AUGMENTING PHOSPHORYLATION OF THE TRANSCRIPTION FACTOR NRF2

Background: Oxidative stress mediates cyclosporine-A (CsA) associated vascular injury post-cardiac transplantation. We investigated the effect of CsA on the Nrf2 pathway by examining vascular function, Nrf2 phosphorylation and superoxide dismutase (SOD) activity and the ability of tBHQ to rescue CsA-mediated injury. Methods and Results: Lewis rats received tBHQ (50mg/kg), CsA (5mg/kg) ± tBHQ, or Saline (CON). Thoracic aortic segments were assessed for vascular function as measured by endothelial-dependent (Edep) relaxation (ED50), and sensitivity to endothelin-1 (ET-1) vasoconstriction. We also analyzed Nrf2 protein expression, phospho-Nrf2/Nrf2 ratio, and SOD activity. CsA significantly impaired Edep vasorelaxation (ED50: 3.5x10-8 +/- 0.4M vs CON 1.8x10-8 +/- 0.5M, p<0.001). tBHQ did not affect vasorelaxation compared to CON (2.18x10-8 +/- 0.4M, p=NS). However, tBHQ improved vasorelaxation in CsA-treated rats significantly (ED50: 2.69x10-8 +/- 0.3M vs CsA alone, p<0.001). Compared to CON, CsA exposure demonstrated increased sensitivity to ET-1 vasospasm (CsA ED50 1.9x10-9 +/- 0.1M vs 2.85x10-9 +/- 0.4M CON, p<0.001) which was rescued significantly by tBHQ exposure compared to CsA alone (ED50: 2.55x10-9 +/- 0.5M). tBHQ alone did not affect sensitivity to ET-1 vasospasm compared to CON (ED50: 2.65x10-9 +/- 0.3M, p=NS). CsA treatment resulted in decreased Nrf2 expression, phosphorylation, and SOD activity (Fig1). Exposure to tBHQ rescued CsA-mediated Nrf2 downregulation, and reduction in Nrf2 phosphorylation and SOD activity (Fig1). Conclusions: Our study suggests potential therapeutic strategies to prevent coronary vascular dysfunction as combined therapy with tBHQ completely abrogated CsA-induced impairment of Nrf2- signalling and vascular injury. Mechanistically, tBHQ may act to rescue Nrf2 expression, phosphorylation and SOD activity in order to ameliorate CsA-mediated coronary dysfunction. ![][1] [1]: /embed/graphic-1.gif