Kaori Nakanishi

Involvement of Endothelial Apoptosis Underlying Chronic Obstructive Pulmonary Disease―like Phenotype in Adiponectin-null Mice: Implications for Therapy

RATIONALE Chronic obstructive pulmonary disease is frequently complicated with comorbidities, such as cardiovascular disease, osteoporosis, and body weight loss, but the causal link remains unclear. OBJECTIVES To investigate the role of adiponectin in the pathogenesis of chronic obstructive pulmonary disease and its potential use in therapy. METHODS Adiponectin localization and dynamics in the lung were analyzed in an elastase-induced emphysema model. Next, the lung of adiponectin-knockout mice, extrapulmonary effects, and the underlying mechanism were investigated. Finally, we tested whether exogenous adiponectin could ameliorate the emphysematous change in adiponectin-knockout mice. MEASUREMENTS AND MAIN RESULTS Adiponectin expression in lung vasculature and plasma concentration of adiponectin were reduced after elastase-instillation. Notably, adiponectin-knockout mice showed progressive alveolar enlargement and increased lung compliance. They further exhibited not only systemic inflammation, but also extrapulmonary phenotype, such as body weight loss, fat atrophy, and osteoporosis. Moreover, endothelial apoptosis was enhanced in the lungs of adiponectin-knockout mice, as evidenced by caspase-3 activity. Consistent with this, expressions of vascular endothelial growth factor receptor-2 and platelet endothelial cell adhesion molecule-1 on endothelial cells were decreased in the adiponectin-knockout mice. Finally, adenovirus-mediated adiponectin supplementation ameliorated the emphysematous phenotype. CONCLUSIONS Adiponectin-knockout mice develop progressive chronic obstructive pulmonary disease-like phenotype with systemic inflammation and extrapulmonary phenotypes. Hypoadiponectinemia could thus play a critical role in the progression of chronic obstructive pulmonary disease and concomitant comorbidities through endothelial dysfunction. Together, adiponectin could be a novel target for chronic obstructive pulmonary disease therapy.

Deletion of tetraspanin CD9 diminishes lymphangiogenesis in vivo and in vitro.

Background: The molecular mechanisms regulating lymphangiogenesis remain unclear. Results: Tetraspanin CD9 modulates molecular organization of integrins in LEC, thereby supporting several functions required for lymphangiogenesis. Conclusion: Deletion of CD9 diminished lymphangiogenesis in mice and humans. Significance: Given that CD9 mediates inflammation and tumor progression, CD9 might be a key component not only in tumor metastasis, but also in inflammation. Tetraspanins have emerged as key players in malignancy and inflammatory diseases, yet little is known about their roles in angiogenesis, and nothing is known about their involvement in lymphangiogenesis. We found here that tetraspanins are abundantly expressed in human lymphatic endothelial cells (LEC). After intrathoracic tumor implantation, metastasis to lymph nodes was diminished and accompanied by decreased angiogenesis and lymphangiogenesis in tetraspanin CD9-KO mice. Moreover, lymphangiomas induced in CD9-KO mice were less pronounced with decreased lymphangiogenesis compared with those in wild-type mice. Although mouse LEC isolated from CD9-KO mice showed normal adhesion, lymphangiogenesis was markedly impaired in several assays (migration, proliferation, and cable formation) in vitro and in the lymphatic ring assay ex vivo. Consistent with these findings in mouse LEC, knocking down CD9 in human LEC also produced decreased migration, proliferation, and cable formation. Immunoprecipitation analysis demonstrated that deletion of CD9 in LEC diminished formation of functional complexes between VEGF receptor-3 and integrins (α5 and α9). Therefore, knocking down CD9 in LEC attenuated VEGF receptor-3 signaling, as well as downstream signaling such as Erk and p38 upon VEGF-C stimulation. Finally, double deletion of CD9/CD81 in mice caused abnormal development of lymphatic vasculature in the trachea and diaphragm, suggesting that CD9 and a closely related tetraspanin CD81 coordinately play an essential role in physiological lymphangiogenesis. In conclusion, tetraspanin CD9 modulates molecular organization of integrins in LEC, thereby supporting several functions required for lymphangiogenesis.

Statins Decrease Lung Inflammation in Mice by Upregulating Tetraspanin CD9 in Macrophages

Tetraspanins organize protein complexes in tetraspanin-enriched membrane microdomains that are distinct from lipid rafts. Our previous studies suggested that reduction in the levels of tetraspanins CD9 and CD81 may be involved in the progression of inflammatory lung diseases, especially COPD. To search for agents that increase the levels of these tetraspanins, we screened 1,165 drugs in clinical use and found that statins upregulate CD9 and CD81 in RAW264.7 macrophages. The lipophilic statins, fluvastatin and simvastatin, reversed LPS-induced downregulation of CD9 and CD81, simultaneously preventing TNF-α and matrix metalloproteinase-9 production and spreading of RAW264.7 cells. These statins exerted anti-inflammatory effects in vitro in wild-type macrophages but not in CD9 knockout macrophages, and decreased lung inflammation in vivo in wild-type mice but not in CD9 knockout mice, suggesting that their effects are dependent on CD9. Mechanistically, the statins promoted reverse transfer of the LPS-signaling mediator CD14 from lipid rafts into CD9-enriched microdomains, thereby preventing LPS receptor formation. Finally, upregulation of CD9/CD81 by statins was related to blockade of GTPase geranylgeranylation in the mevalonate pathway. Our data underscore the importance of the negative regulator CD9 in lung inflammation, and suggest that statins exert anti-inflammatory effects by upregulating tetraspanin CD9 in macrophages.

Etanercept improved primary biliary cirrhosis associated with rheumatoid arthritis

are surgery, presence of a foreign body, trauma, immunodeficiency, diabetes mellitus, malignancy and sickle cell disease [1]. Only five cases of Peptostreptococcus spp. spondylodiscitis have been reported [4e7] (Table 1). The lack of its recovery in practice may be attributable to improper methods for sample collection, transport and culture (contamination of the sample, exposure to oxygen or desiccation). Most Peptostreptococcus spp. isolates (96%) are susceptible to b-lactam antibiotics [3], although, resistances have been reported against ticarcillin and cefotaxime. Clindamycin and metronidazole are slightly less effective: about 85% of isolates are susceptible [3]. Quinolones, except newer fluoroquinolones as moxifloxacin [8], and aminosides have poor bactericidal activity [3]. When an appropriate antibiotic drug is used, prognosis of spondylodiscitis is the same as with other germs. This case enlights, as recommended by Grados et al. [9], the need for microbial identification and antibiotic susceptibility testing before antibiotic administration. Indeed, bacteria as Peptostreptococcus spp. can be resistant to metronidazole or clindamycin suggested for anaerobic discitis treatment [9].

Adiponectin: A Novel Link Between Adipocytes and COPD

Adiponectin (APN) is a unique adipokine with multiple salutary effects such as antiapoptotic, anti-inflammatory, and anti-oxidative activities in numerous organs and cells. Chronic obstructive pulmonary disease (COPD), a growing cause of mortality and morbidity worldwide, often results from the smoking habit and is considered a lifestyle-related disease. COPD is frequently complicated with comorbidities, such as cardiovascular disease, diabetes mellitus, and osteoporosis; however, the molecular mechanisms linking COPD and the associated comorbidities are poorly understood. Recent data have revealed a role for APN in the lung; mice lacking APN spontaneously develop a COPD-like phenotype with extrapulmonary effects, including systemic inflammation, body weight loss, and osteoporosis. This finding highlights the key role of APN in lung pathology and the novel cross talk between lung and adipose tissues. This review summarizes recent advances in understanding the physiological and pathological role of APN in the lung.

Klotho-related Molecules Upregulated by Smoking Habit in Apparently Healthy Men: A Cross-sectional Study

While aging is unavoidable, the aging mechanism is still unclear because of its complexity. Smoking causes premature death and is considered as an environmental aging accelerator. In the present study, we focused on the influence of smoking to the serum concentration of anti-aging protein α-klotho (αKl) and the β-klotho-associated protein fibroblast growth factor (FGF)-21 in men. Subjects consisted of apparently healthy men over 40 years of age who underwent health examination. Physical and biochemical parameters, including the levels of several cytokines and growth factors, were obtained from the subjects. Among middle-aged men (46.1 ± 5.1 years), serum levels of FGF-21, soluble αKl (sαKl), and inflammation-related cytokine interleukin (IL)-6 were significantly higher in smokers than in never-smokers. Serum levels of FGF-21 increased and correlated with alanine transaminase, γ guanosine-5′-triphosphate, and total cholesterol only in smokers, suggesting FGF-21 as a metabolic disorder-related factor in smokers. In aged men (60.3 ± 1.7 years), although the serum levels of sαKl in never-smokers were low, smokers showed highly increased serum levels of sαKl. Serum levels of sαKl was correlated with IL-6 in middle-aged never-smokers, suggesting sαKl regulates IL-6. However, this correlation was disrupted in smokers and aged men.

Gender differences in the relationship between blood pressure and body mass index during adolescence.

OBJECTIVE In adults, gender and obesity play significant roles in the regulation of blood pressure (BP). This study investigated the effects of gender and body mass index (BMI) on BP during adolescence. DESIGN AND SETTING A cross-sectional and longitudinal study involving 6838 students under twenty years old (median, eighteen years old; male, 4624; female, 2214) at Osaka University visited the Healthcare Center for their matriculation health examination from April to May in the years 2008, 2009, and 2010, and re-visited the Healthcare Center for their student health examination from May to June in the years 2011, 2012, and 2013. METHODS Height, body weight, and BP were measured in students both on and 3 years after admission to Osaka University. RESULTS On admission, the slope of the regression line for BMI and systolic BP (SBP) in non-underweight students was significantly different between genders. SBP and diastolic BP (DBP) increased in both genders during the observation period. Among male students who had a normal BMI on admission, those who had an increase in BMI of over 4% during the observation period showed a greater increase in SBP than those who had a change in BMI of -4% to 4%. On the other hand, female students showed no change in BP with the increase in BMI. CONCLUSIONS The magnitude of BP elevation with increased BMI was associated with gender during adolescence. This may be a cause of the higher prevalence of hypertension in adult males.

Association between N-terminal pro-brain natriuretic peptide and adiponectin in healthy Japanese men.

BACKGROUND The natriuretic peptides, brain natriuretic peptide (BNP) and N-terminal-proBNP (NT-proBNP), are cardiac-derived hormones and can serve as biomarkers for ventricular dysfunction. BNP has cardio-protective effects and is known as a regulator of metabolism. In the present study, to evaluate the relationship between natriuretic peptides and metabolic disorders, we focused on the association between NT-proBNP and metabolic syndrome-related molecule adiponectin (APN). METHODS Forty-five apparently healthy men who underwent health examination at the Osaka University Health Care Center were enrolled for this study. Physical and biochemical parameters including serum APN and NT-proBNP concentrations were obtained from all the subjects. RESULTS The serum concentrations of NT-proBNP negatively correlated with metabolic disorder parameters, body mass index (BMI), waist circumferences, and fasting plasma glucose levels, but positively correlated with APN, suggesting that similar to APN, NT-proBNP is associated with metabolic disorders. Furthermore, increased serum concentrations of APN were found to be accompanied by increased serum concentrations of NT-proBNP and decreased BMI and mean intima-media thickness. CONCLUSIONS The serum concentrations of NT-proBNP are associated with APN concentrations and metabolic disorder parameters in healthy subjects.

Validation of Noninvasive Morphological and Diffusion Imaging in Mouse Emphysema by Micro–Computed Tomography and Hyperpolarized 129Xe Magnetic Resonance Imaging

Animal disease models are pivotal in investigating the pathogenesis of emphysema and developing novel drugs, but the modalities to evaluate murine emphysema models have been of limited validity and sensitivity. In this study, we evaluated hyperpolarized (129)Xe magnetic resonance imaging (MRI) and micro-computed tomography (micro-CT) compared with traditional methods, such as plethysmography and histology. Elastase-treated mice and adiponectin knockout mice were used as murine emphysema models to evaluate these modalities. Three weeks after elastase administration, significant and heterogeneous emphysema was evaluated according to the mean linear intercept and plethysmography parameters. Notably, the distribution of low-density areas, as examined by micro-CT, correlated with the mean linear intercept and plethysmography parameters in whole lungs. These correlations were also observed in regional areas. Furthermore, we introduced hyperpolarized (129)Xe MRI, which can evaluate gas exchange between the alveoli and blood during spontaneous breathing. Parameters of gas exchange (fD) and alveolar size (Vs/Va) were significantly decreased in elastase-treated mice, and moderately correlated with the plethysmography parameters. Of importance, we could detect a decrease of the fD value in low-density areas with micro-CT, suggesting that gas exchange decreased in emphysematous lesions. Likewise, these parameters (fD and Vs/Va) were also decreased in adiponectin knockout mice, which exhibit emphysema with a homogeneous distribution. We demonstrated the feasibility of (129)Xe MRI and micro-CT in combination with traditional modalities. These noninvasive modalities provide complementary data that can be used for repeated estimations of regional gas exchange and lung morphology.

Double deletion of tetraspanins CD9 and CD81 in mice leads to a syndrome resembling accelerated aging

Chronic obstructive pulmonary disease (COPD) has been recently characterized as a disease of accelerated lung aging, but the mechanism remains unclear. Tetraspanins have emerged as key players in malignancy and inflammatory diseases. Here, we found that CD9/CD81 double knockout (DKO) mice with a COPD-like phenotype progressively developed a syndrome resembling human aging, including cataracts, hair loss, and atrophy of various organs, including thymus, muscle, and testis, resulting in shorter survival than wild-type (WT) mice. Consistent with this, DNA microarray analysis of DKO mouse lungs revealed differential expression of genes involved in cell death, inflammation, and the sirtuin-1 (SIRT1) pathway. Accordingly, expression of SIRT1 was reduced in DKO mouse lungs. Importantly, siRNA knockdown of CD9 and CD81 in lung epithelial cells additively decreased SIRT1 and Foxo3a expression, but reciprocally upregulated the expression of p21 and p53, leading to reduced cell proliferation and elevated apoptosis. Furthermore, deletion of these tetraspanins increased the expression of pro-inflammatory genes and IL-8. Hence, CD9 and CD81 might coordinately prevent senescence and inflammation, partly by maintaining SIRT1 expression. Altogether, CD9/CD81 DKO mice represent a novel model for both COPD and accelerated senescence.