In this study, we examined the biological action of IL-17 on human non-small cell lung cancer (NSCLC). Although IL-17 had no direct effect on the in vitro growth rate of NSCLC, IL-17 selectively augmented the secretion of an array of angiogenic CXC chemokines, including CXCL1, CXCL5, CXCL6, and CXCL8 but not angiostatic chemokines, by three different NSCLC lines. Endothelial cell chemotactic activity (as a measure of net angiogenic potential) was increased in response to conditioned medium from NSCLC stimulated with IL-17 compared with those from unstimulated NSCLC. Enhanced chemotactic activity was suppressed by neutralizing mAb(s) to CXCL1, CXCL5, and CXCL8 or to CXCR-2 but not to vascular endothelial growth factor-A. Transfection with IL-17 into NSCLC had no effect on the in vitro growth, whereas IL-17 transfectants grew more rapidly compared with controls when transplanted in SCID mice. This IL-17-elicited enhancement of NSCLC growth was associated with increased tumor vascularity. Moreover, treatment with anti-mouse CXCR-2-neutralizing Ab significantly attenuated the growth of both neomycin phosphotransferase gene-transfected and IL-17-transfected NSCLC tumors in SCID mice. A potential role for IL-17 in modulation of the human NSCLC phenotype was supported by the findings that, in primary NSCLC tissues, IL-17 expression was frequently detected in accumulating and infiltrating inflammatory cells and that high levels of IL-17 expression were associated with increased tumor vascularity. These results demonstrate that IL-17 increases the net angiogenic activity and in vivo growth of NSCLC via promoting CXCR-2-dependent angiogenesis and suggest that targeting CXCR-2 signaling may be a novel promising strategy to treat patients with NSCLC.

Background-Persistently high plasma endothelin-1 (ET-1) levels in diabetic patients have been associated with the development of cardiac fibrosis, which results from the deposition of extracellular matrix and fibroblast recruitment from an as-yet unknown source. The underlying mechanism, however, remains elusive. Here, we hypothesize that ET-1 might contribute to the accumulation of cardiac fibroblasts through an endothelial-to-mesenchymal transition in diabetic hearts. Methods and Results-We induced diabetes mellitus in vascular endothelial cell-specific ET-1 knockout [ET-1 f/f ;Tie2-Cre (ϩ)] mice and their wild-type littermates using the toxin streptozotocin. Gene expression and histological and functional parameters were examined at 8, 24, and 36 weeks after the induction of diabetes mellitus. Diabetes mellitus increased cardiac ET-1 expression in wild-type mice, leading to mitochondrial disruption and myofibril disarray through the generation of superoxide. Diabetic mice also showed impairment of cardiac microvascularization and a decrease in cardiac vascular endothelial growth factor expression. ET-1 further promotes cardiac fibrosis and heart failure through the accumulation of fibroblasts via endothelial-to-mesenchymal transition. All of these features were abolished in ET-1 f/f ;Tie2-Cre (ϩ) hearts. Targeted ET-1 gene silencing by small interfering RNA in cultured human endothelial cells ameliorated high glucose-induced phenotypic transition and acquisition of a fibroblast marker through the inhibition of transforming growth factor-␤ signaling activation and preservation of the endothelial cell-to-cell contact regulator VE-cadherin. Conclusions-These results provide new insights suggesting that diabetes mellitus-induced cardiac fibrosis is associated with the emergence of fibroblasts from endothelial cells and that this endothelial-to-mesenchymal transition process is stimulated by ET-1. Targeting endothelial cell-derived ET-1 might be beneficial in the prevention of diabetic cardiomyopathy. (Circulation. 2010;121:2407-2418.)Key Words: cardiomyopathy Ⅲ diabetes mellitus Ⅲ endothelin Ⅲ fibrosis Ⅲ heart failure D iabetes mellitus can affect cardiac structure and function, and this may lead to heart failure in the absence of coronary atherosclerosis and hypertension. However, despite the importance of this clinical entity, the multifactorial nature of the disease remains incompletely understood. Three decades have passed since Rubler et al 1 first described patients with diabetic cardiomyopathy, and to date, a number of epidemiological, clinical, and experimental studies have confirmed the existence of this unique cardiomyopathy. 2,3 Several mechanisms are considered to be important in the development of cardiac structural and ultrastructural changes, with hyperglycemia and altered cardiac metabolism being proposed as central to the pathophysiology of this disorder. 4 Given the increased risk of heart failure and cardiovascular events in diabetic patients, a better understanding of the underlying mechanisms...

Estrogen-related receptor ␣ (ERR␣) was identified as a gene related to estrogen receptor ␣ (ER␣) and belongs to a class of nuclear orphan receptors. ERR␣ binds to estrogen responsive element(s) (ERE) and is considered to be involved in modulation of estrogenic actions. However, biological significance of ERR␣ remains largely unknown. Therefore, we examined the expression of ERR␣ in human breast carcinoma tissues using immunohistochemistry (n ‫؍‬ 102) and real-time reverse transcription-PCR (n ‫؍‬ 30). ERR␣ immunoreactivity was detected in the nuclei of carcinoma cells in 55% of breast cancers examined, and relative immunoreactivity of ERR␣ was significantly (P ‫؍‬ 0.0041) associated with the mRNA level. Significant associations were detected between ER␣ and ERE-containing estrogen-responsive genes, such as pS2 (P < 0.0001) and EBAG9/RCAS1 (P ‫؍‬ 0.0214), in breast carcinoma tissues. However, no significant association was detected between ER␣ and pS2 (P ‫؍‬ 0.1415) in the ERR␣-positive cases (n ‫؍‬ 56) or between ER␣ and EBAG9/RCAS1 (P ‫؍‬ 0.8271) in the ERR␣-negative group (n ‫؍‬ 46). ERR␣ immunoreactivity was significantly associated with an increased risk of recurrence and adverse clinical outcome by both uni-(P ‫؍‬ 0.0097 and P ‫؍‬ 0.0053, respectively) and multi-(P ‫؍‬ 0.0215 and P ‫؍‬ 0.0118, respectively) variate analyses. A similar tendency was also detected in the group of breast cancer patients who received tamoxifen therapy after surgery. Results from our study suggest that ERR␣ possibly modulates the expression of ERE-containing estrogen-responsive genes, and ERR␣ immunoreactivity is a potent prognostic factor in human breast carcinoma.

Purpose: The possible involvement of gender-dependent factors has been suggested in human non-small cell lung carcinomas (NSCLC), but their precise roles remain largely unclear. Therefore, we examined intratumoral estradiol concentrations in NSCLC to examine local actions of estrogens in NSCLC. Experimental Design: Fifty-nine frozen specimens of NSCLC were available for liquid chromatography/electrospray tandem mass spectrometry to study intratumoral estradiol concentrations. In addition, A549 NSCLC cells stably expressing estrogen receptor (ER) a (A549 + ERa) or ERh (A549 + ERh) were used in vitro studies. Results: Forty-three (73%) of 59 NSCLC showed higher concentration of estradiol in carcinoma tissues than the corresponding nonneoplastic lung tissues from the same patient, and intratumoral estradiol concentrations were significantly (P = 0.0002 and 2.2-fold) higher than the corresponding nonneoplastic lungs. The intratumoral concentration of estradiol was positively correlated with aromatase expression, tumor size, and Ki-67 status in ERa-or ERh-positive cases. In in vitro studies, estradiol significantly increased cell proliferation of A549 + ERa orA549 + ERh, which was significantly suppressed by selective ER modulators, tamoxifen or raloxifene. Both A549 + ERa and A549 + ERh cells expressed aromatase. The cell proliferation level in these cells was significantly increased under treatment with testosterone, and it was inhibited by addition of the aromatase inhibitor letrozole. Conclusions: These results suggest that estradiol is locally produced in NSCLC mainly by aromatase and plays an important role in the growth of ERa-or ERh-positive NSCLC. Therefore, use of selective ER modulators and/or aromatase inhibitors may be clinically effective in NSCLC that are positive for both ER and aromatase.

Diabetic kidney disease is a major cause of renal failure that urgently necessitates a breakthrough in disease management. Here we show using untargeted metabolomics that levels of phenyl sulfate, a gut microbiota-derived metabolite, increase with the progression of diabetes in rats overexpressing human uremic toxin transporter SLCO4C1 in the kidney, and are decreased in rats with limited proteinuria. In experimental models of diabetes, phenyl sulfate administration induces albuminuria and podocyte damage. In a diabetic patient cohort, phenyl sulfate levels significantly correlate with basal and predicted 2-year progression of albuminuria in patients with microalbuminuria. Inhibition of tyrosine phenol-lyase, a bacterial enzyme responsible for the synthesis of phenol from dietary tyrosine before it is metabolized into phenyl sulfate in the liver, reduces albuminuria in diabetic mice. Together, our results suggest that phenyl sulfate contributes to albuminuria and could be used as a disease marker and future therapeutic target in diabetic kidney disease.