Depressed adrenomedullin in the embryonic transforming growth factor-beta1 null mouse becomes elevated postnatally

Transforming growth factor-beta (TGF-beta) and adrenomedullin are multifunctional regulatory proteins which are expressed in developing embryonic and adult tissues. Because of their colocalization, TGF-beta1 and adrenomedullin may be able to coordinately act to influence development and differentiation. In order to learn more about the biology of adrenomedullin in the absence of the effects of TGF-beta1 in vivo, we examined adrenomedullin in the TGF-beta1 null mouse. A generally lower amount of adrenomedullin was detected by immunohistochemical staining analysis in multiple tissues from embryonic TGF-beta1 null mice compared to wildtype animals, including the heart, lung, brain, liver, and kidney, among others. In contrast, immunohistochemical staining for adrenomedullin was more intense in tissues of the postnatal TGF-beta1 null mouse compared to the wildtype mouse. These observations were confirmed by quantitative real time RT-PCR for adrenomedullin in both embryos and postnatal animals, as well as in cultured mouse embryo fibroblasts from TGF-beta1 null and wildtype mice. In addition, when cultured mouse embryo fibroblasts were treated with a neutralizing monoclonal antibody against TGF-beta1, the levels of adrenomedullin expression were statistically reduced compared to untreated cells. Our data show that expression of adrenomedullin is reduced in tissues of the developing embryonic TGF-beta1 null mouse compared to the wildtype mouse, but increases during postnatal development in TGF-beta1 null mice. The elevated expression of adrenomedullin which occurs postnatally in the TGF-beta1 null mouse may be a cause or a consequence of the multifocal wasting syndrome which is characteristic of postnatal TGF-beta1 null mice

TMPRSS4 regulates levels of integrin α5 in NSCLC through miR-205 activity to promote metastasis

TMPRSS4 is a membrane-anchored protease involved in cell migration and invasion in different cancer types including lung cancer. TMPRSS4 expression is increased in NSCLC and its inhibition through shRNA reduces lung metastasis. However, molecular mechanisms leading to the protumorigenic regulation of TMPRSS4 in lung cancer are unknown. METHODS: miR-205 was identified as an overexpressed gene upon TMPRSS4 downregulation through microarray analysis. Cell migration and invasion assays and in vivo lung primary tumour and metastasis models were used for functional analysis of miR-205 overexpression in H2170 and H441 cell lines. Luciferase assays were used to identify a new miR-205 direct target in NSCLC. RESULTS: miR-205 overexpression promoted an epithelial phenotype with increased E-cadherin and reduced fibronectin. Furthermore, miR-205 expression caused a G0/G1 cell cycle arrest and inhibition of cell growth, migration, attachment to fibronectin, primary tumour growth and metastasis formation in vivo. Integrin α5 (a proinvasive protein) was identified as a new miR-205 direct target in NSCLC. Integrin α5 downregulation in lung cancer cells resulted in complete abrogation of cell migration, a decreased capacity to adhere to fibronectin and reduced in vivo tumour growth, compared with control cells. TMPRSS4 silencing resulted in a concomitant reduction of integrin α5 levels. CONCLUSION: We have demonstrated for the first time a new molecular pathway that connects TMPRSS4 and integrin α5 through miR-205 to regulate cancer cell invasion and metastasis. Our results will help designing new therapeutic strategies to inhibit this novel pathway in NSCLC

Effect of Systemic Hypertension With Versus Without Left Ventricular Hypertrophy on the Progression of Atrial Fibrillation (from the Euro Heart Survey).

Hypertension is a risk factor for both progression of atrial fibrillation (AF) and development of AF-related complications, that is major adverse cardiac and cerebrovascular events (MACCE). It is unknown whether left ventricular hypertrophy (LVH) as a consequence of hypertension is also a risk factor for both these end points. We aimed to assess this in low-risk AF patients, also assessing gender-related differences. We included 799 patients from the Euro Heart Survey with nonvalvular AF and a baseline echocardiogram. Patients with and without hypertension were included. End points after 1 year were occurrence of AF progression, that is paroxysmal AF becoming persistent and/or permanent AF, and MACCE. Echocardiographic LVH was present in 33% of 379 hypertensive patients. AF progression after 1 year occurred in 10.2% of 373 patients with rhythm follow-up. In hypertensive patients with LVH, AF progression occurred more frequently as compared with hypertensive patients without LVH (23.3% vs 8.8%, p = 0.011). In hypertensive AF patients, LVH was the most important multivariably adjusted determinant of AF progression on multivariable logistic regression (odds ratio 4.84, 95% confidence interval 1.70 to 13.78, p = 0.003). This effect was only seen in male patients (27.5% vs 5.8%, p = 0.002), while in female hypertensive patients, no differences were found in AF progression rates regarding the presence or absence of LVH (15.2% vs 15.0%, p = 0.999). No differences were seen in MACCE for hypertensive patients with and without LVH. In conclusion, in men with hypertension, LVH is associated with AF progression. This association seems to be absent in hypertensive women

Progression From Paroxysmal to Persistent Atrial Fibrillation. Clinical Correlates and Prognosis

Objectives: We investigated clinical correlates of atrial fibrillation (AF) progression and evaluated the prognosis of patients demonstrating AF progression in a large population. Background: Progression of paroxysmal AF to more sustained forms is frequently seen. However, not all patients will progress to persistent AF. Methods: We included 1,219 patients with paroxysmal AF who participated in the Euro Heart Survey on AF and had a known rhythm status at follow-up. Patients who experienced AF progression after 1 year of follow-up were identified. Results: Progression of AF occurred in 178 (15%) patients. Multivariate analysis showed that heart failure, age, previous transient ischemic attack or stroke, chronic obstructive pulmonary disease, and hypertension were the only independent predictors of AF progression. Using the regression coefficient as a benchmark, we calculated the HATCH score. Nearly 50% of the patients with a HATCH score >5 progressed to persistent AF compared with only 6% of the patients with a HATCH score of 0. During follow-up, patients with AF progression were more often admitted to the hospital and had more major adverse cardiovascular events. Conclusions: A substantial number of patients progress to sustained AF within 1 year. The clinical outcome of these patients regarding hospital admissions and major adverse cardiovascular events was worse compared with patients demonstrating no AF progression. Factors known to cause atrial structural remodeling (age and underlying heart disease) were independent predictors of AF progression. The HATCH score may help to identify patients who are likely to progress to sustained forms of AF in the near future. \ua9 2010 American College of Cardiology Foundation

Estudio de la expresión de ghrelina en píloro, duodeno y yeyuno de ratones NOD y NOD/SCID de 20 semanas

La diabetes mellitus (DM) es una enfermedad crónica caracterizada por una hiperglucemia derivada de una disminución en la secreción y/o de una deficiencia en la acción de la insulina. Una de las características clínicas de la DM es el retraso en el vaciamiento gástrico, también conocido como gastroparesia diabética. La motilidad gastrointestinal está regulada por varias hormonas gastrointestinales, entre las que se encuentra la ghrelina, cuya función es la estimulación del vaciamiento gástrico. Es probable que en la DM, la expresión de esta hormona se vea disminuida, generando la gastroparesia diabética. En este trabajo se ha estudiado la expresión de ghrelina en el píloro, duodeno y yeyuno de 20 ratones (machos y hembras) tipo NOD (modelo animal de DM1) y 10 ratones macho tipo NOD/SCID (grupo control) de 20 semanas de edad, mediante una técnica inmunocitoquímica (ICQ). Además, los ratones NOD se han dividido en función de sus glucemias (8 ratones NOD normoglucémicos (NODN), 8 ratones NOD hiperglucémicos (NODH) y 4 ratones NOD diabéticos (NODD)). Tras la realización de la técnica ICQ, se calculó la densidad de células inmunorreactivas (IR) para la ghrelina, tanto en la túnica mucosa del píloro y yeyuno, como en la túnica mucosa y submucosa del duodeno. Los resultados obtenidos, en todos los grupos, muestran una disminución significativa de la densidad de células IR para la ghrelina desde el píloro hacia el tracto gastrointestinal inferior. Se ha encontrado un aumento no significativo de la densidad celular IR para la ghrelina en el píloro y duodeno de los ratones NOD hembra frente a los ratones NOD macho y NOD/SCID y un aumento significativo en el yeyuno de los ratones NOD macho frente al grupo control. Por último, se ha observado una disminución no significativa de la densidad de células IR para la ghrelina en el píloro y duodeno de los ratones NODD frente a los NODN y NODH

Estudio de la expresión de ghrelina en píloro, duodeno y yeyuno de ratones NOD y NOD/SCID de 20 semanas

La diabetes mellitus (DM) es una enfermedad crónica caracterizada por una hiperglucemia derivada de una disminución en la secreción y/o de una deficiencia en la acción de la insulina. Una de las características clínicas de la DM es el retraso en el vaciamiento gástrico, también conocido como gastroparesia diabética. La motilidad gastrointestinal está regulada por varias hormonas gastrointestinales, entre las que se encuentra la ghrelina, cuya función es la estimulación del vaciamiento gástrico. Es probable que en la DM, la expresión de esta hormona se vea disminuida, generando la gastroparesia diabética. En este trabajo se ha estudiado la expresión de ghrelina en el píloro, duodeno y yeyuno de 20 ratones (machos y hembras) tipo NOD (modelo animal de DM1) y 10 ratones macho tipo NOD/SCID (grupo control) de 20 semanas de edad, mediante una técnica inmunocitoquímica (ICQ). Además, los ratones NOD se han dividido en función de sus glucemias (8 ratones NOD normoglucémicos (NODN), 8 ratones NOD hiperglucémicos (NODH) y 4 ratones NOD diabéticos (NODD)). Tras la realización de la técnica ICQ, se calculó la densidad de células inmunorreactivas (IR) para la ghrelina, tanto en la túnica mucosa del píloro y yeyuno, como en la túnica mucosa y submucosa del duodeno. Los resultados obtenidos, en todos los grupos, muestran una disminución significativa de la densidad de células IR para la ghrelina desde el píloro hacia el tracto gastrointestinal inferior. Se ha encontrado un aumento no significativo de la densidad celular IR para la ghrelina en el píloro y duodeno de los ratones NOD hembra frente a los ratones NOD macho y NOD/SCID y un aumento significativo en el yeyuno de los ratones NOD macho frente al grupo control. Por último, se ha observado una disminución no significativa de la densidad de células IR para la ghrelina en el píloro y duodeno de los ratones NODD frente a los NODN y NODH

Spatial-temporal protein expression of inhibitor of differentiation-1 (Id1) during fetal embryogenesis and in different mouse and human cancer types

Inhibitor of differentiation-1 (Id1) plays a role in cell proliferation, acquisition of epithelial to mesenchymal transition (EMT) features and angiogenesis. Id1 was shown to be expressed in some tumor types, mainly in advanced dedifferentiated stages. However, recent studies using a validated and highly specific monoclonal antibody against Id1 have challenged many of the results obtained by immunohistochemistry. The goal of our work was to perform a thorough analysis of Id1 expression in mouse embryos and adult tissues, as well as healthy and malignant mouse and human samples using this validated antibody (Perk et al., 2006). Our results show that Id1 was highly expressed in the oropharyngeal cavity, lung, cartilage and skin of E14 and E15 mouse embryos, but expression was progressively reduced in more developed embryos. Immunostaining only remained in epithelial cells of the gut and uterus of adult mice. Mammary MMTV-Myc and MMTV-Myc/VEGF transgenic mouse tumors, and squamous cell carcinomas of the lung induced by N-nitroso-tris-chloroethylurea (NTCU) were highly positive for Id1, unlike their respective healthy counterparts. Id1 immunostaining in a human tissue microarray (TMA) revealed strong expression in cancers of the oral cavity, bladder and cervix. Some tumor specimens of esophagus, thyroid and breast were also strongly positive. Our results suggest that Id1 is an oncofetal protein highly expressed in particular tumor types that should be reanalyzed in future studies using large cohorts of patients to reassess its diagnostic/prognostic value. Moreover, MMTV-Mycand NTCU-induced tumors could serve as appropriate mouse models to study Id1 functions in breast and lung cancer, respectively

TMPRSS4 regulates levels of integrin α5 in NSCLC through miR-205 activity to promote metastasis

TMPRSS4 is a membrane-anchored protease involved in cell migration and invasion in different cancer types including lung cancer. TMPRSS4 expression is increased in NSCLC and its inhibition through shRNA reduces lung metastasis. However, molecular mechanisms leading to the protumorigenic regulation of TMPRSS4 in lung cancer are unknown. METHODS: miR-205 was identified as an overexpressed gene upon TMPRSS4 downregulation through microarray analysis. Cell migration and invasion assays and in vivo lung primary tumour and metastasis models were used for functional analysis of miR-205 overexpression in H2170 and H441 cell lines. Luciferase assays were used to identify a new miR-205 direct target in NSCLC. RESULTS: miR-205 overexpression promoted an epithelial phenotype with increased E-cadherin and reduced fibronectin. Furthermore, miR-205 expression caused a G0/G1 cell cycle arrest and inhibition of cell growth, migration, attachment to fibronectin, primary tumour growth and metastasis formation in vivo. Integrin α5 (a proinvasive protein) was identified as a new miR-205 direct target in NSCLC. Integrin α5 downregulation in lung cancer cells resulted in complete abrogation of cell migration, a decreased capacity to adhere to fibronectin and reduced in vivo tumour growth, compared with control cells. TMPRSS4 silencing resulted in a concomitant reduction of integrin α5 levels. CONCLUSION: We have demonstrated for the first time a new molecular pathway that connects TMPRSS4 and integrin α5 through miR-205 to regulate cancer cell invasion and metastasis. Our results will help designing new therapeutic strategies to inhibit this novel pathway in NSCLC