Yoichi Nishii

Deficiency of tenascin C attenuates allergen-induced bronchial asthma in the mouse

Tenascin C (TN‐C) is an extracellular matrix glycoprotein whose expression is increased in several inflammatory diseases of the lung, including bronchial asthma. However, the exact function of TN‐C in the pathogenesis of lung inflammation remains unclear. In the present study, we compared the degree of bronchial asthma in wild‐type and TN‐C‐deficient (–/–) BALB/c mice. Bronchial asthma was induced by sensitization and challenge with ovalbumin. Littermates treated with saline were used as controls. Cytokines in bronchoalveolar lavage fluid and plasma were measured by enzyme immunoassays. The number of eosinophils in the lung was significantly increased in wild‐type mice compared with TN‐C‐knockout mice. Airway hyperreactivity, NF‐κB activation and concentrations of monocyte chemoattractant protein‐1, IL‐5, IL‐13, metalloproteinase‐9 and immunoglobulin‐E in the bronchoalveolar lavage fluid were significantly decreased in ovalbumin‐sensitized/challenged TN‐C‐knockout mice compared with their wild‐type counterparts. In vitro experiments disclosed that TN‐C significantly stimulates the secretion of IL‐5, IL‐13, IFN‐γ and immunoglobulin‐E from spleen lymphocytes. These observations suggest that TN‐C is involved in the pathogenesis of bronchial asthma.

Decreased expression of aquaporin-5 in bleomycin-induced lung fibrosis in the mouse

The expression of aquaporin‐5, the major water channel expressed in alveolar, tracheal, and upper bronchial epithelium, is significantly down‐regulated during acute lung injury. In the present study, the expression of aquaporin‐5 in two different mouse models of lung fibrosis was evaluated. Lung fibrosis was induced by intratracheal and by subcutaneous infusion of bleomycin. The expression of aquaporin‐5 was investigated by immunohistochemical studies and by polymerase chain reaction. There were many cells with loss of aquaporin‐5 immunoreactivity in type I alveolar epithelial cells in the mouse models of lung fibrosis. Immunohistochemistry of lung tissue in aquaporin‐5 knockout mice revealed a fibrotic phenotype with increased deposition of extracellular collagen type I in thickened alveolar walls. Semiquantitative analysis of aquaporin‐5 mRNA expression showed more abundant content of aquaporin‐5 in the lung of the normal mouse compared to the mouse with lung fibrosis. The results of this study showed, for the first time, that chronic lung injury and lung fibrosis is associated with decreased protein and mRNA expression of aquaporin‐5 in the lung.

Protective role of protein C inhibitor in monocrotaline-induced pulmonary hypertension

Summary.  Background: Protein C inhibitor (PCI) plays a role in multiple biological processes including fertilization, coagulation, fibrinolysis and kinin systems. Objectives: We hypothesized that PCI participates in the pathogenesis of pulmonary hypertension. To demonstrate this, we compared the development of pulmonary hypertension in mice overexpressing PCI in the lung with wild‐type (WT) mice. Pulmonary hypertension was induced by s.c. injection of 600 mg kg−1 of monocrotaline weekly for 8 weeks. Results: Right ventricular arterial pressure was significantly increased in monocrotaline‐treated WT mice compared with that in monocrotaline‐treated transgenic mice. Bronchoalveolar lavage fluid (BALF) levels of thrombin–antithrombin complex, monocyte chemoattractant protein‐1 and platelet‐derived growth factor, and the plasma level of tumor necrosis factor‐α were significantly increased in monocrotaline‐treated WT mice as compared with monocrotaline‐treated PCI transgenic mice. Increased level of PCI‐thrombin complex was detected in BALF from monocrotaline‐treated PCI transgenic mice as compared with saline‐treated PCI transgenic mice. Conclusions: This study showed that increased expression of PCI in the lung is protective against monocrotaline‐induced pulmonary hypertension, suggesting a potential beneficial effect of PCI for the therapy of this disease.

Smart watch‑based coaching with tiotropium and olodaterol ameliorates physical activity in patients with chronic obstructive pulmonary disease

Combined therapy with tiotropium and olodaterol notably improves parameters of lung function and quality of life in patients with chronic obstructive pulmonary disease (COPD) compared to mono-components; however, its effect on physical activity is unknown. The present study evaluated whether combination therapy affects daily physical performance in patients with COPD under a smart watch-based encouragement program. This was a non-blinded clinical trial with no randomization or placebo control. A total of 20 patients with COPD were enrolled in the present study. The patients carried an accelerometer for 4 weeks; they received no therapy during the first 2 weeks but they were treated with combined tiotropium and olodaterol under a smart watch-based encouragement program for the last 2 weeks. The pulmonary function test, COPD assessment test, 6-min walk distance and parameters of physical activity were significantly improved (P<0.05) by combination therapy under smart watch-based coaching compared with values prior to treatment. To the best of our knowledge, the present study for the first time provides evidence that smart watch-based coaching in combination with tiotropium and olodaterol may improve daily physical activity in chronic obstructive pulmonary disease.

Utility of Liquid Biopsy by Improved PNA-LNA PCR Clamp Method for Detecting EGFR Mutation at Initial Diagnosis of Non–Small-Cell Lung Cancer: Observational Study of 190 Consecutive Cases in Clinical Practice

Micro‐Abstract We reviewed 190 consecutive unselected patients who underwent liquid biopsy for detecting an EGFR mutation. The results indicated that not all patients should be candidates for liquid biopsy at initial diagnosis, while some patients gain benefit from initial liquid biopsy in clinical practice. These real‐world data supply useful information in choosing patients appropriate for liquid biopsy. Background: The clinical benefit of liquid biopsy for unselected patients at initial diagnosis has thus far been unclear. We aimed to evaluate the utility of liquid biopsy at initial diagnosis, as well as the efficacy of epidermal growth factor receptor–tyrosine kinase inhibitor (EGFR‐TKI) based on liquid biopsy results in clinical practice, using the improved peptide nucleic acid–locked nucleic acid (PNA‐LNA) PCR clamp method. Patients and Methods: We routinely performed liquid biopsy using the improved PNA‐LNA PCR clamp method for all patients diagnosed with non–small‐cell lung cancer (NSCLC) between June 2015 and October 2016. We retrospectively evaluated the reliability of liquid biopsy based either on clinical stage or between sensitizing EGFR mutation and T790M mutation, and the clinical benefit of EGFR‐TKI based on the liquid biopsy results in practice. Results: A total of 244 patients underwent liquid biopsies, with 168 patients tested at diagnosis and 22 tested for T790M after pretreatment of EGFR‐TKI. For detecting a sensitizing EGFR mutation, the sensitivity, specificity, positive predictive value, and negative predictive value were 72.7%, 100%, 100%, and 93.7% in the group with advanced‐stage NSCLC and 0, 100%, not evaluable, and 70.5% in the group with early‐stage NSCLC. The positive predictive value and negative predictive value for T790M were 33.3% and 55.6%, respectively. Fourteen patients in the liquid‐positive group and 16 patients in the tissue‐positive group received EGFR‐TKI. The objective response rates of first‐ and second‐generation EGFR‐TKI for the liquid‐positive and tissue‐positive groups were 90.0% and 90.9%, respectively. There was no significant difference in median progression‐free survival between the liquid‐positive and tissue‐positive groups (P = .839). Conclusion: Patients with early‐stage NSCLC should not be candidates for this liquid biopsy method. We recommend tissue biopsy as the preferred initial method of molecular analysis, with the exception of patients who are T790M positive or patients who are unable to tolerate invasive biopsy. Graphical abstract: Figure. No Caption available.

Comparable immunoreactivity rates of PD-L1 in archival and recent specimens from non-small cell lung cancer: PD-L1 in archival and recent samples

Molecular targeted therapy including the use of monoclonal antibodies directed against the immune checkpoints PD‐L1 and PD‐1 receptor have remarkably improved the therapeutic response and survival of cancer patients. The tumor expression level of PD‐L1 can predict the response rate to checkpoint inhibitors. We evaluated whether the time interval between tumor tissue sampling/paraffinization and immunohistochemistry affects the staining level of PD‐L1 in non‐small cell lung cancer (NSCLC).

Renal Injury during Long-Term Crizotinib Therapy

Crizotinib is highly effective against anaplastic lymphoma kinase-positive and c-ros oncogen1-positive non-small cell lung cancer. Renal dysfunction is associated with crizotinib therapy but the mechanism is unknown. Here, we report a case of anaplastic lymphoma kinase positive non-small cell lung cancer showing multiple cysts and dysfunction of the kidneys during crizotinib administration. We also present results demonstrating that long-term crizotinib treatment induces fibrosis and dysfunction of the kidneys by activating the tumor necrosis factor-α/nuclear factor-κB signaling pathway. In conclusion, this study shows the renal detrimental effects of crizotinib, suggesting the need of careful monitoring of renal function during crizotinib therapy.

Efficacy of osimertinib in a patient with non‑small cell lung cancer harboring epithelial growth factor receptor exon 19 deletion/T790M mutation, with poor performance status

Osimertinib, a third-generation epithelial growth factor receptor (EGFR) tyrosine kinase inhibitor, has been demonstrated to be effective for treating patients with T790M-positive advanced non-small cell lung cancer (NSCLC) with a relatively good performance status (grade 0–1). Reports of therapeutic response to osimertinib in advanced NSCLC patients with poor performance status are infrequent. The present case report discusses a patient with advanced lung adenocarcinoma harboring EGFR exon 19 deletion and T790M mutation with central nervous system involvement and poor performance status. The patient had a past history of partial lung resection due to lung adenocarcinoma, positive genetic test for EGFR exon 19 deletion in post-surgical tumor specimens, and therapy with erlotinib and onartuzumab for the appearance of a lung metastatic tumor during the post-surgical follow-up. The combined therapy was continued until the discovery of metastatic tumors in bones and the central nervous system. The Cobas test performed using tissue from bone metastatic tumor was positive for exon 19 deletion and for T790M mutation. The patient was treated with osimertinib and adverse effects or hematological toxicity were not observed. Performance status of the patient improved from grade 4 to 2. Subsequent studies revealed remission of bone metastasis and reduced central nervous system lesions. This report provides evidence on the safety and efficacy of osimertinib for treating NSCLC patients with progressive disease, central nervous system lesion and poor performance status.