Researchers Database

hagiwara kouichi

    InternalMedicinePulmonary Professor
Last Updated :2021/11/23

Researcher Information


  • MD, PhD(University of Tokyo)

J-Global ID

Research Interests

  • 肺癌   GeneChip   TGF-β   全ゲノムSNP解析   HGF   ホモ接合指紋法   増殖抑制   SLPI   疾患遺伝子解析   細胞株   HM on zHH   EGFR   非小細胞肺癌   GWS+   elafin   特発性肺線維症   ホモ接合ハプロタイプ法   アポートシス   ホモ接合ハプロタイプ   変異遺伝子   薬剤性肺障害   近親婚   Smad4   SNP   肺胞微石症   急性増悪   p53   染色体20番   耐性   c-ab1   

Research Areas

  • Life sciences / Respiratory medicine
  • Life sciences / Urology

Academic & Professional Experience

  • 2015  埼玉医科大学医学部教授

Published Papers

  • Mizushina Yoshiko, Karasawa Tadayoshi, Aizawa Kenichi, Kimura Hiroaki, Watanabe Sachiko, Kamata Ryo, Komada Takanori, Mato Naoko, Kasahara Tadashi, Koyama Shinichiro, Bando Masashi, Hagiwara Koichi, Takahashi Masafumi
    JOURNAL OF IMMUNOLOGY 203 (1) 236 - 246 0022-1767 2019/07 [Refereed][Not invited]
    Inflammation plays a pivotal role in the pathophysiology of gastric aspiration-induced acute lung injury (ALI). However, its mechanism remains unclear. In this study, we investigated the role of NLRP3 inflammasome-driven IL-1β production in a mouse model of acid aspiration-induced inflammation and ALI. Acid aspiration-induced inflammatory responses and ALI in wild-type mice were significantly attenuated in IL-1β-/- mice, but not NLRP3-/- mice. In vitro experiments revealed that severe acidic stress (pH 1.75) induced the processing of pro-IL-1β into its 18-kDa mature form (p18-IL-1β), which was different from the caspase-1-processed 17-kDa form (p17-IL-1β), in human THP-1 macrophages and primary murine macrophages. Deficiency of NLRP3 and caspase-1 had no effect on acidic stress-produced IL-1β. The production of IL-1β by severe acidic stress was prevented by inhibitors of serine proteases [4-(2-aminoethyl)benzenesulfonyl fluoride hydrochloride], but not of cysteine proteases (E-64), cathepsin G, or inflammasome. The cathepsin D inhibitor pepstatin A inhibited IL-1β production induced by mild acidic stress (pH 6.2) or lactic acid, but not severe acidic stress. Using mass spectrometry and processing-site mutants of pro-IL-1β, we identified D109 as a novel cleavage site of pro-IL-1β in response to severe acidic stress and calculated the theoretical molecular mass of the mature form to be 18.2 kDa. The bioactivity of acidic stress-produced IL-1β was confirmed by its ability to promote p38 phosphorylation and chemokine upregulation in alveolar epithelial cells. These findings demonstrate a novel mechanism of acid-induced IL-1β production and inflammation independent of NLRP3 inflammasome and provide new insights into the therapeutic strategies for aspiration pneumonitis and ALI.
  • Haruhiro Saito, Tatsuro Fukuhara, Naoki Furuya, Kana Watanabe, Shunichi Sugawara, Shunichiro Iwasawa, Yoshio Tsunezuka, Ou Yamaguchi, Morihito Okada, Kozo Yoshimori, Ichiro Nakachi, Akihiko Gemma, Koichi Azuma, Futoshi Kurimoto, Yukari Tsubata, Yuka Fujita, Hiromi Nagashima, Gyo Asai, Satoshi Watanabe, Masaki Miyazaki, Koichi Hagiwara, Toshihiro Nukiwa, Satoshi Morita, Kunihiko Kobayashi, Makoto Maemondo
    The Lancet. Oncology 20 (5) 625 - 635 1470-2045 2019/05 [Refereed][Not invited]
    BACKGROUND: Resistance to first-generation or second-generation EGFR tyrosine kinase inhibitor (TKI) monotherapy develops in almost half of patients with EGFR-positive non-small-cell lung cancer (NSCLC) after 1 year of treatment. The JO25567 phase 2 trial comparing erlotinib plus bevacizumab combination therapy with erlotinib monotherapy established the activity and manageable toxicity of erlotinib plus bevacizumab in patients with NSCLC. We did a phase 3 trial to validate the results of the JO25567 study and report here the results from the preplanned interim analysis. METHODS: In this prespecified interim analysis of the randomised, open-label, phase 3 NEJ026 trial, we recruited patients with stage IIIB-IV disease or recurrent, cytologically or histologically confirmed non-squamous NSCLC with activating EGFR genomic aberrations from 69 centres across Japan. Eligible patients were at least 20 years old, and had an Eastern Cooperative Oncology Group performance status of 2 or lower, no previous chemotherapy for advanced disease, and one or more measurable lesions based on Response Evaluation Criteria in Solid Tumours (1.1). Patients were randomly assigned (1:1) to receive oral erlotinib 150 mg per day plus intravenous bevacizumab 15 mg/kg once every 21 days, or erlotinib 150 mg per day monotherapy. Randomisation was done by minimisation, stratified by sex, smoking status, clinical stage, and EGFR mutation subtype. The primary endpoint was progression-free survival. This study is ongoing; the data cutoff for this prespecified interim analysis was Sept 21, 2017. Efficacy was analysed in the modified intention-to-treat population, which included all randomly assigned patients who received at least one dose of treatment and had at least one response evaluation. Safety was analysed in all patients who received at least one dose of study drug. The trial is registered with the University Hospital Medical Information Network Clinical Trials Registry, number UMIN000017069. FINDINGS: Between June 3, 2015, and Aug 31, 2016, 228 patients were randomly assigned to receive erlotinib plus bevacizumab (n=114) or erlotinib alone (n=114). 112 patients in each group were evaluable for efficacy, and safety was evaluated in 112 patients in the combination therapy group and 114 in the monotherapy group. Median follow-up was 12·4 months (IQR 7·0-15·7). At the time of interim analysis, median progression-free survival for patients in the erlotinib plus bevacizumab group was 16·9 months (95% CI 14·2-21·0) compared with 13·3 months (11·1-15·3) for patients in the erlotinib group (hazard ratio 0·605, 95% CI 0·417-0·877; p=0·016). 98 (88%) of 112 patients in the erlotinib plus bevacizumab group and 53 (46%) of 114 patients in the erlotinib alone group had grade 3 or worse adverse events. The most common grade 3-4 adverse event was rash (23 [21%] of 112 patients in the erlotinib plus bevacizumab group vs 24 [21%] of 114 patients in the erlotinib alone group). Nine (8%) of 112 patients in the erlotinib plus bevacizumab group and five (4%) of 114 patients in the erlotinib alone group had serious adverse events. The most common serious adverse events were grade 4 neutropenia (two [2%] of 112 patients in the erlotinib plus bevacizumab group) and grade 4 hepatic dysfunction (one [1%] of 112 patients in the erlotinib plus bevacizumab group and one [1%] of 114 patients in the erlotinib alone group). No treatment-related deaths occurred. INTERPRETATION: The results of this interim analysis showed that bevacizumab plus erlotinib combination therapy improves progression-free survival compared with erlotinib alone in patients with EGFR-positive NSCLC. Future studies with longer follow-up, and overall survival and quality-of-life data will be required to further assess the efficacy of this combination in this setting. FUNDING: Chugai Pharmaceutical.
  • Takigami A, Yamasawa H, Kurosaki A, Sakamoto N, Onuki T, Mato N, Tetsuka K, Endo S, Niki T, Bando M, Hagiwara K
    Internal medicine (Tokyo, Japan) 58 (9) 1335 - 1339 0918-2918 2019/05 [Refereed][Not invited]
  • Inoue A, Yamaguchi T, Tanaka K, Sakashita A, Aoe K, Seki N, Hagiwara K
    Internal medicine (Tokyo, Japan) 58 (10) 1399 - 1403 0918-2918 2019/05 [Refereed][Not invited]
  • Kurniawan FD, Alia D, Priyanto H, Mahdani W, Hagiwara K
    Respiratory investigation 2212-5345 2019/04 [Refereed][Not invited]
  • Kurosaki Fumio, Takemura Tamiko, Bando Masashi, Kuroki Tomonori, Numao Toshio, Moriyama Hiroshi, Hagiwara Koichi
    BMC PULMONARY MEDICINE 19 1471-2466 2019/01 [Refereed][Not invited]
  • Onuki Tsugitoshi, Nakayama Masayuki, Bando Masashi, Yada Rie, Sekine Toshie, Yamauchi Hiroyoshi, Hisata Shu, Nagai Yoshiaki, Suzuki Takuji, Hagiwara Koichi
    RESPIROLOGY 23 275  1323-7799 2018/11 [Refereed][Not invited]
  • Kawaguchi Makoto, Nakayama Masayuki, Bando Masashi, Dotake Yoichi, Mato Naoko, Yamasawa Hideaki, Suzuki Takuji, Hagiwara Koichi, Sugiyama Yukihiko
    RESPIROLOGY 23 240 - 241 1323-7799 2018/11 [Refereed][Not invited]
  • Hirahara K, Mato N, Hagiwara K, Nakayama T
    Journal of leukocyte biology 104 (5) 895 - 901 0741-5400 2018/11 [Refereed][Not invited]
  • Fumio Kurosaki, Ryosuke Uchibori, Yoshihide Sehara, Yasushi Saga, Masashi Urabe, Hiroaki Mizukami, Koichi Hagiwara, Akihiro Kume
    Human gene therapy 29 (11) 1242 - 1251 1043-0342 2018/11 [Refereed][Not invited]
    Idiopathic pulmonary fibrosis (IPF) is a fibroproliferative disorder with limited therapeutic options. An aberrant wound healing process in response to repetitive lung injury has been suggested for its pathogenesis, and a number of cytokines including transforming growth factor β1 play pivotal roles in the induction and progression of fibrosis. Thus, the regulation of these pro-inflammatory conditions may reduce the progression of IPF and ameliorate its symptoms in patients. Interleukin-10 (IL-10), a pleiotropic cytokine, exerts anti-inflammatory and anti-fibrotic effects in numerous biological settings. In the present study, we investigated the preventive effects of IL-10 on bleomycin-induced pulmonary fibrosis in mice with the continuous expression of this cytokine via an adeno-associated virus serotype 6 vector. Mice were administered the adeno-associated virus serotype 6 vector encoding mouse IL-10 by intratracheal injection, and osmotic minipumps containing bleomycin were subcutaneously implanted seven days later. Lung histology and the expression levels of pro-inflammatory cytokines and fibrogenic cytokines were then analyzed. In mice exhibiting persistent IL-10 expression on day 35, the number of infiltrated inflammatory cells and the development of fibrosis in lung tissues were significantly reduced. Increases in transforming growth factor β1 and decreases in IFN-γ were also suppressed in treated animals, with changes in these cytokines playing important roles in the pathogenesis of pulmonary fibrosis. Furthermore, IL-10 significantly improved survival in bleomycin-induced mice. Our results provide insights into the potential benefit of the anti-fibrotic effects of IL-10 as a novel therapeutic approach for IPF.
  • Nakayama M, Yamamoto S, Kaneko N, Mato N, Suzuki T, Hagiwara K
    Respirology case reports 6 (7) e00351  2051-3380 2018/10 [Refereed][Not invited]
  • Saito S, Lasky JA, Hagiwara K, Kondoh Y
    Respiratory investigation 56 (5) 375 - 383 2212-5345 2018/09 [Refereed][Not invited]
  • Tsugitoshi Onuki, Eri Morita, Noritaka Sakamoto, Yoshiaki Nagai, Masafumi Sata, Koichi Hagiwara
    Respirology Case Reports 6 (6) e00334  2051-3380 2018/08 [Refereed][Not invited]
    Pembrolizumab is an immune checkpoint inhibitor that induces side effects called “immune-related adverse events” (irAEs). Various types of organs are affected by irAEs, although reports of upper gastrointestinal disorders are rare. Here, we report a case of extensive inflammatory pathologies in the oesophagus, stomach, duodenum, and jejunum after the administration of pembrolizumab for non-small cell lung cancer.
  • Kudo F, Watanabe Y, Iwai Y, Miwa C, Nagai Y, Ota H, Yabe H, Demitsu T, Hagiwara K, Koyama N, Koyama S
    Internal medicine (Tokyo, Japan) 57 (15) 2217 - 2221 0918-2918 2018/08 [Refereed][Not invited]
  • Iwai Yuki, Kikuchi Ryota, Watanabe Yasutaka, Koyama Nobuyuki, Hagiwara Koichi, Nakamura Hiroyuki, Aoshiba Kazutetsu
    CANCER RESEARCH 78 (13) 0008-5472 2018/07 [Refereed][Not invited]
  • Matsubara Daisuke, Soda Manabu, Yoshimoto Taichiro, Amano Yusuke, Kihara Atsushi, Sakuma Yuji, Endo Shunsuke, Hagiwara Koichi, Fukayama Masashi, Mano Hiroyuki, Niki Toshiro
    CANCER RESEARCH 78 (13) 0008-5472 2018/07 [Refereed][Not invited]
  • Fumio Kurosaki, Taichiro Yoshimoto, Masayuki Nakayama, Masashi Bando, Koichi Hagiwara
    Journal of Infection and Chemotherapy 24 (6) 483 - 486 1437-7780 2018/06 [Refereed][Not invited]
    Pulmonary infection due to Mycobacterium heckeshornense (M. heckeshornense) in healthy adults without underlying diseases is very rare and optimal treatments have not yet been established. A 39-year-old woman was admitted to our hospital for further examinations following the identification of a pulmonary cavitary nodule. Acid-fast bacilli were cultured from specimens obtained by bronchofiberscopy, and identified with M. heckeshornense using nucleotide sequencing. Antimycobacterial chemotherapy was effective temporarily, while the nodular lesion subsequently worsened. The patient underwent lobectomy and has not relapsed thus far. A lung specimen showed marked granulomatous inflammation with extensive caseous necrosis and the preservation of some parts of alveolar septa within caseous necrosis, indicating an exudative process and resistance to chemotherapy. M. heckeshornense is strongly pathogenic and switching to surgical intervention needs to be considered when chemotherapy is insufficient.
  • Oizumi S, Sugawara S, Minato K, Harada T, Inoue A, Fujita Y, Maemondo M, Watanabe S, Ito K, Gemma A, Demura Y, Fukumoto S, Isobe H, Kinoshita I, Morita S, Kobayashi K, Hagiwara K, Aiba K, Nukiwa T
    ESMO open 3 (2) e000313  2059-7029 2018/02 [Refereed][Not invited]
    Background: The North-East Japan Study Group (NEJ) 005/Tokyo Cooperative Oncology Group (TCOG) 0902 study has reported that first-line concurrent and sequential alternating combination therapies of an epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor (gefitinib) plus platinum-based doublet chemotherapy (carboplatin/pemetrexed) offer promising efficacy with predictable toxicities for patients with EGFR-mutant non-small cell lung cancer. However, overall survival (OS) data were insufficient in the primary report because of the lack of death events. Patients and methods: Progression-free survival (PFS) and OS were re-evaluated at the final data cut-off point (March 2017) for the entire population (n=80). Results: At the median follow-up time of 35.6 months, 88.8% of patients had progressive disease and 77.5% of patients had died. Median PFS was 17.5 months for the concurrent regimen and 15.3 months for the sequential alternating regimen (P=0.13). Median OS was 41.9 and 30.7 months, respectively (P=0.036). Updated response rates were similar in both groups (90.2% and 82.1%, respectively; P=0.34). Patients with Del19 tumours displayed relatively better OS (median: 45.3 vs 33.3 months, respectively) than those with L858R (31.4 vs 28.9 months, respectively). No severe adverse events, including interstitial lung disease, occurred in the period since the primary report. Conclusions: This updated analysis confirms that PFS is improved with first-line combination therapy compared with gefitinib monotherapy and that the concurrent regimen, in particular, offers an OS benefit of 42 months in the EGFR-mutated setting. Our ongoing NEJ009 study will clarify whether this combination strategy can be incorporated into routine clinical practice. Trial registration number: UMIN C000002789, Post-results.
  • Kiyoshi Hirahara, Naoko Mato, Tomomi Ichikawa, Jin Kumagai, Masayuki Nakayama, Hideaki Yamasawa, Masashi Bando, Koichi Hagiwara, Yukihiko Sugiyama, Toshinori Nakayama
    CYTOKINE 100 69 - 69 1043-4666 2017/12 [Refereed][Not invited]
  • Takeo Nakaya, Hisashi Oshiro, Ayako Takigami, Yoshihiko Kanai, Kenji Tetsuka, Koichi Hagiwara, Hirofumi Fujii, Shunsuke Endo, Akira Tanaka
    Medicine (United States) 96 (50) e8926  1536-5964 2017/12 [Refereed][Not invited]
    Rationale: Solitary fibrous tumors are mesenchymal tumors presenting as fibroblastic neoplasms with prominent branching vascular patterns, which are often generated from the pleura. Most solitary fibrous tumors are benign however, some can turn malignant. High-grade sarcomas from solitary fibrous tumors include multidirectional histopathological components. Patient concerns: We describe our experience of a giant high-grade sarcoma with mixed components generated from a solitary fibrous tumor of the pleura in a 67-year-old female patient presenting with cough and left-sided chest pain. The patient had been diagnosed with a pleural mass in the left chest by X-ray about 30 years earlier. However, the tumor was allowed to grow, without surgical intervention, for a long time. Interventions: Thoracic surgeons performed the removal of the giant pleural tumor the tumor measured 18.0 × 14.5 × 10 cm in size, and was considered a giant tumor generated from the pleura of the left chest cavity. Diagnoses: The surgically removed tumor was solid and light brownish, and included myxoid and arabesque pattern lesions. The tumor also showed hemorrhagic and necrotic lesions. Moreover, spindle cells with less atypia, resembling fibroblasts, were noted. These spindle tumor cells were CD34- and Stat6-positive, suggesting a solitary fibrous tumor. Some of the spindle tumor cells were surrounded by thick collagenous fibers. Considering that the tumor originated from the parietal pleura, the tumor was defined as a solitary fibrous tumor in origin. The tumor also comprised high-grade sarcomatous components these included lipid-rich, rhabdomyosarcomatous, and pleomorphic components. The high-grade sarcoma component included bizarre tumor cells with severe atypia. Outcomes: Tumor recurrence occurred in the left chest about 4 months after the surgery, and the patient died 8 months postoperatively. Lessons: The present case clearly demonstrates that a solitary fibrous tumor can develop into high-grade sarcomatous overgrowth, including lipid-rich, rhabdomyosarcoma, and pleomorphic sarcoma components, if left untreated for a prolonged period. This case provides profound insights about the natural history, histogenesis, differentiation, and malignant transformation of solitary fibrous tumors.
  • Akiko Okuyama, Hideaki Yamasawa, Hiroyoshi Yamauchi, Masayuki Nakayama, Naoko Mato, Masashi Bando, Koichi Hagiwara
    RESPIROLOGY 22 200 - 200 1323-7799 2017/11 [Refereed][Not invited]
  • Hiroyoshi Yamauchi, Masayuki Nakayama, Shinichi Yamamoto, Tatsuya Saito, Akiko Okuyama, Naoko Mato, Hideaki Yamasawa, Takuji Suzuki, Masashi Bando, Koichi Hagiwara
    RESPIROLOGY 22 242 - 242 1323-7799 2017/11 [Refereed][Not invited]
  • Hideaki Yamasawa, Michiru Sawahata, Masayuki Nakayama, Masaki Toshima, Masashi Bando, Koichi Hagiwara
    RESPIROLOGY 22 261 - 261 1323-7799 2017/11 [Refereed][Not invited]
  • Ferry Dwi Kurniawan, Herry Priyanto, Wilda Mahdani, Dina Alia, Koichi Hagiwara
    RESPIROLOGY 22 269 - 269 1323-7799 2017/11 [Refereed][Not invited]
  • Yasuhiro Kondoh, Hiroyuki Taniguchi, Kensuke Kataoka, Taiki Furukawa, Masahiko Ando, Kenta Murotani, Michiaki Mishima, Yoshikazu Inoue, Takashi Ogura, Masashi Bando, Koichi Hagiwara, Takafumi Suda, Hirofumi Chiba, Hiroki Takahashi, Yukihiko Sugiyama, Sakae Homma
    RESPIROLOGY 22 (8) 1609 - 1614 1323-7799 2017/11 [Refereed][Not invited]
    Background and objective: In Japan, the classification of disease severity of idiopathic pulmonary fibrosis (IPF) (J-system) has been used in making decisions on medical care subsidies. The present J-system consists of arterial partial pressure of oxygen (PaO2) and exercise desaturation in stages of I-IV. It provides a good prognostic classification in stages III and IV, but not in stages I and II. Therefore, we propose a revised system to improve discriminative ability in stages I and II. Methods: We compared the revised J-system with the present J-system using Cox proportional hazards model to predict mortality rate. We also evaluated the recently proposed GAP (Gender, Age and Physiology) system in comparison to both J-systems. Results: Two-hundred and fifteen IPF patients were studied retrospectively. A univariate model showed that the present and revised J-systems and a modified GAP system were all significant prognostic factors. The C-statistic for discriminating prognosis was higher in the revised J-system than the modified GAP system and the present J-system (0.677, 0.652 and 0.659, respectively). The C-statistics of these models produced from the 10 000 bootstrap samples were similar to those of the original models, suggesting good internal validation (0.665 (95% CI: 0.621-0.705), 0.645 (0.600-0.686) and 0.659 (0.616-0.700), respectively). Multivariate analysis revealed that the revised J-system (P = 0.0038) and the modified GAP system (P = 0.0029) were independent prognostic factors. Conclusion: The revised J-system can provide a better mortality prediction than the present one. Both the revised J-system and the modified GAP system are independent and valuable tools for prognostication and clinical management for IPF.
  • Baohui Han, Sergei Tjulandin, Koichi Hagiwara, Nicola Normanno, Laksmi Wulandari, Konstantin Laktionov, Achmad Hudoyo, Yong He, Yi-Ping Zhang, Meng-Zhao Wang, Chien Ying Liu, Marianne Ratcliffe, Rose McCormack, Martin Reck
    LUNG CANCER 113 37 - 44 0169-5002 2017/11 [Refereed][Not invited]
    Objectives: Limited understanding exists of epidermal growth factor receptor (EGFR) mutation frequency in less common subgroups of advanced non-small-cell lung cancer (aNSCLC) (e.g. squamous cell carcinoma [SCC]), and to what extent local practices exclude patients from EGFR testing based on their clinical characteristics. Materials and methods: IGNITE (non-comparative/-interventional; NCT01788163) was conducted in 90 centres (Asia-Pacific/Russia). Eligible patients: local/metastatic aNSCLC; chemotherapy-naive, newly-diagnosed/recurrent disease after resection; ineligible for curative treatment. Patients provided a tissue/cytology (all) and a blood plasma (China/Russia/South Korea/Taiwan) sample. Primary endpoint: EGFR mutation frequency in aNSCLC patients (adenocarcinoma [ADC]/non-ADC), as per local practices. Results: 3382 patients were enrolled. EGFR mutation frequencies for evaluable tissue/cytology samples in Asia-Pacific and Russian patients: 49.3% (862/1749) and 18.0% (90/500) for ADC tumours; 14.1% (74/525) and 3.7% (15/402) for non-ADC; 9.9% (40/403) and 3.7% (13/349) for SCC. Of Russian patients with SCC tumours harbouring common, activating EGFR mutations, 6/9 were never-/former-smokers. Mutation status concordance between 2581 matched tissue/cytology and plasma samples: 80.5% (sensitivity 46.9%, specificity 95.6%). Conclusion: EGFR mutation testing should be considered in all Asian aNSCLC patients. Also, as activating EGFR mutations were observed in a small number of Caucasian squamous NSCLC patients, testing here may be appropriate, particularly in those with no/remote smoking history. Circulating free tumour-derived DNA is feasible for mutation analysis employing well-validated and sensitive methods, when tumour samples are unavailable.
  • Nakayama Masayuki, Yamamoto Shinichi, Kaneko Naoki, Mato Naoko, Yamasawa Hideaki, Bando Masashi, Hagiwara Koichi
    EUROPEAN RESPIRATORY JOURNAL 50 0903-1936 2017/09 [Refereed][Not invited]
  • Daisuke Matsubara, Manabu Soda, Taichiro Yoshimoto, Yusuke Amano, Yuji Sakuma, Azusa Yamato, Toshihide Ueno, Shinya Kojima, Tomoki Shibano, Yasuyuki Hosono, Masahito Kawazu, Yoshihiro Yamashita, Shunsuke Endo, Koichi Hagiwara, Masashi Fukayama, Takashi Takahashi, Hiroyuki Mano, Toshiro Niki
    Cancer Science 108 (9) 1888 - 1896 1349-7006 2017/09 [Refereed][Not invited]
    The major driver mutations of lung cancer, EGFR mutations and EML4-ALK fusion, are mainly detected in terminal respiratory unit (TRU)-type lung adenocarcinomas, which typically show lepidic and/or papillary patterns, but are rarely associated with a solid or invasive mucinous morphology. In order to elucidate the key genetic events in non-TRU-type lung cancer, we carried out whole-exome sequencing on 43 non-TRU-type lung adenocarcinomas based on morphology (17 acinar, nine solid, and two enteric adenocarcinomas, and 15 adenocarcinomas with a mucinous morphology). Our analysis identified mutations in TP53 (16/43, 37.2%), KRAS (13/43, 30.2%), and NKX2-1/TTF-1 (7/43 16.3%) as the top three significantly mutated genes, while the EGFR mutation was rare (1/43, 2.3%) in this cohort. Eight NKX2-1/TTF-1 mutations (five frameshift, two nonsense, and one missense) were identified, with one case harboring two distinct NKX2-1/TTF-1 mutations (one missense and one frameshift). Functional assays with the NK2 homeobox 1 (NKX2-1)/thyroid transcription factor 1 (TTF-1) mutants revealed that none of them retain the activity as a transcriptional factor. Histologically, invasive mucinous adenocarcinomas accounted for most of the NKX2-1/TTF-1 mutations (five cases), as well as one enteric and one acinar adenocarcinoma. Immunohistochemistry showed that the cohort was largely divided into TTF-1-postive/hepatocyte nuclear factor 4-α (HNF4-α)-negative and TTF-1-negative/HNF4-α-positive groups. NKX2-1/TTF-1 mutations were exclusively found in the latter, in which the gastrointestinal markers, mucin 5AC and cytokeratin 20, were frequently expressed. Bisulfite sequencing revealed that the NKX2-1/TTF-1 gene body was highly methylated in NKX2-1/TTF-1-negative cases, including those without the NKX2-1/TTF-1 mutations. The genetic or epigenetic inactivation of NKX2-1/TTF-1 may play an essential role in the development and aberrant differentiation of non-TRU-type lung adenocarcinomas.
  • Miyako Satouchi, Hiroshi Tanaka, Hiroshige Yoshioka, Tadasuke Shimokawaji, Keiko Mizuno, Koji Takeda, Ichiro Yoshino, Takashi Seto, Takayasu Kurata, Naoki Tashiro, Koichi Hagiwara
    LUNG CANCER 111 190 - 194 0169-5002 2017/09 [Refereed][Not invited]
    Objective: Detection of epidermal growth factor receptor (EGFR) gene mutations is essential in deciding therapeutic strategy in non-small cell lung cancer (NSCLC) patients at initial diagnosis. Moreover, in EGFR mutation positive (EGFRm) NSCLC patients, re-biopsy at disease progression to clarify resistance mechanisms is also important. However, collecting histology samples is often difficult because of inaccessibility and invasiveness. In some cases, only cytology samples can be collected, and studies have reported that cytology samples are appropriate for EGFR gene mutation testing. The cobas (R) EGFR Mutation Test (Roche Molecular Systems Inc., Branchburg, New Jersey, USA) is approved as a companion diagnostic for osimertinib, a third-generation EGFR-tyrosine kinase inhibitor approved in Japan. However, it is not clear whether the EGFR T790M mutation can be detected in cytology samples using this test. The primary objective of this study was to assess concordance of EGFR T790M gene mutation detection between histology and matched cytology samples using the cobas (R) EGFR Mutation Test. Materials and methods: We conducted a multicenter, observational study in Japan. Overall, 41 EGFRm NSCLC patients who had both histology and cytology samples collected at the same time at re-biopsy and with the results of EGFR mutation test using histology samples were enrolled. The EGFR mutation status of both sample types was tested using the cobas (R) EGFR Mutation Test and the concordance rates were calculated. Results: The EGFR T790M mutation detection rate in histology and cytology samples was 42.5% and 37.5%, respectively. The overall percent agreement between the histology and cytology samples was 91.7%. Conclusions: These data demonstrate that the cobas (R) EGFR Mutation Test can detect the EGFR T790M mutation in both cytology and histology samples.
  • Naoko Mato, Kiyoshi Hirahara, Tomomi Ichikawa, Jin Kumagai, Masayuki Nakayama, Hideaki Yamasawa, Masashi Bando, Koichi Hagiwara, Yukihiko Sugiyama, Toshinori Nakayama
    SCIENTIFIC REPORTS 7 (1) 6805  2045-2322 2017/07 [Refereed][Not invited]
    The lung develops an unique epithelial barrier system to protect host from continuous invasion of various harmful particles. Interleukin (IL-)33 released from epithelial cells in the lung drives the type 2 immune response by activating ST2-expressed immune cells in various allergic diseases. However, the involvement of memory-type ST2(+)CD4(+)T cells in such lung inflammation remains unclear. Here we demonstrated that intratracheal administration of IL-33 resulted in the substantial increase of numbers of tissue-resident memory-type ST2(+)CD4(+)T cells in the lung. Following enhanced production of IL-5 and IL-13, eosinophilic lung inflammation sequentially developed. IL-33-mediated eosinophilic lung inflammation was not fully developed in T cell-deficient Foxn1(nu) mice and NSG mice. Dexamethasone treatment showed limited effects on both the cell number and function of memory-type ST2(+)CD4(+)T cells. Thus our study provides novel insight into the pathogenesis of eosinophilic lung disease, showing that memory-type ST2(+)CD4(+)T cells are involved in IL-33-induced eosinophilic inflammation and elicited steroid-resistance.
  • Rintaro Noro, Kunihiko Kobayashi, Jiro Usuki, Yukio Hosomi, Masaru Nishitsuji, Hiroaki Okamoto, Mitsunori Hino, Koichi Hagiwara, Akihiko Miyanaga, Masahiro Seike, Kaoru Kubota, Akihiko Gemma
    JOURNAL OF CLINICAL ONCOLOGY 35 0732-183X 2017/05 [Refereed][Not invited]
  • Yoshiaki Inoue, Jun Shiihara, Hitoshi Miyazawa, Hiromitsu Ohta, Megumi Higo, Yoshiaki Nagai, Kunihiko Kobayashi, Yasuo Saijo, Masanori Tsuchida, Mitsuo Nakayama, Koichi Hagiwara
    PLOS ONE 12 (4) e0176525  1932-6203 2017/04 [Refereed][Not invited]
    Molecular targeting therapy for non-small cell lung cancer (NSCLC) has clarified the importance of mutation testing when selecting treatment regimens. As a result, multiple-gene mutation tests are urgently needed. We developed a next-generation sequencer (NGS)-based, multi-gene test named the MINtS for investigating driver mutations in both cytological specimens and snap-frozen tissue samples. The MINtS was used to investigate the EGFR, KRAS, BRAF genes from DNA, and the ERBB2, and the ALK, ROS1, and RET fusion genes from RNA. We focused on high specificity and sensitivity (>= 0.99) and even included samples with a cancer cell content of 1%. The MINtS enables testing of more than 100 samples in a single run, making it possible to process a large number of samples submitted to a central laboratory, and reducing the cost for a single sample. We investigated 96 cytological samples and 190 surgically resected tissues, both of which are isolated in daily clinical practice. With the cytological samples, we compared the results for the EGFR mutation between the MINtS and the PNA-LNA PCR clamp test, and their results were 99% consistent. In the snap-frozen tissue samples, 188/190 (99%) samples were successfully analyzed for all genes investigated using both DNA and RNA. Then, we used 200 cytological samples that were serially isolated in clinical practice to assess RNA quality. Using our procedure, 196 samples (98%) provided high-quality RNA suitable for analysis with the MINtS. We concluded that the MINtS test system is feasible for analyzing ''druggable'' genes using cytological samples and snap-frozen tissue samples. The MINtS will fill a needs for patients for whom only cytological specimens are available for genetic testing.
  • Takakiyo Nakaya, Masashi Bando, Koichi Hagiwara, Yukihiko Sugiyama
    RHEUMATOLOGY 56 140 - 140 1462-0324 2017/03 [Refereed][Not invited]
  • Masayuki Ito, Kazuyuki Nakagome, Hiromitsu Ohta, Keiichi Akasaka, Yoshitaka Uchida, Atsushi Hashimoto, Ayako Shiono, Toshinori Takada, Makoto Nagata, Jun Tohyama, Koichi Hagiwara, Minoru Kanazawa, Koh Nakata, Ryushi Tazawa
    BMC PULMONARY MEDICINE 17 (1) 40  1471-2466 2017/02 [Refereed][Not invited]
    Background: Pulmonary alveolar proteinosis (PAP) is a rare lung disease characterized by surfactant accumulation, and is caused by disruption of granulocyte/macrophage colony-stimulating factor (GM-CSF) signaling. Abnormalities in CSF2 receptor alpha (CSF2RA) were reported to cause pediatric hereditary PAP. We report here the first case of CSF2RA-mutated, elderly-onset hereditary (h) PAP. Case presentation: The patient developed dyspnea on exertion, and was diagnosed with PAP at the age of 77 years, based on findings from chest computed tomography scan and bronchoalveolar lavage. She tested negative for GM-CSF autoantibodies, with no underlying disease. Her serum GM-CSF level was elevated (91.3 pg/mL), indicating GM-CSF signaling impairment and genetic defects in the GM-CSF receptor. GM-CSF-stimulated phosphorylation in signal transducer and activator of transcription 5 (STAT5) was not observed, and GM-CSF-Ra expression was defective in her blood cells. Genetic screening revealed a homozygous, single-base C > T mutation at nt 508-a nonsense mutation that yields a stop codon (Q170X)-in exon 7 of CSF2RA. High-resolution analysis of single nucleotide polymorphism array confirmed a 22.8-Mb loss of heterozygosity region in Xp22.33p22.11, encompassing the CSF2RA gene. She was successfully treated with whole lung lavage (WLL), which reduced the serum levels of interleukin (IL)-2, IL-5, and IL-17, although IL-3 and M-CSF levels remained high. Conclusions: This is the first known report of elderly-onset hPAP associated with a CSF2RA mutation, which caused defective GM-CSF-Ra expression and impaired signaling. The analyses of serum cytokine levels during WLL suggested that GM-CSF signaling might be compensated by other signaling pathways, leading to elderly-onset PAP.
  • Yoshiaki Inoue, Ato Sugiyama, Kohei Aoki, Hiroki Fukuda, Masatoshi Gika, Yotaro Izumi, Kunihiko Kobayashi, Mitsuo Nakayama, Koichi Hagiwara
    JOURNAL OF THORACIC ONCOLOGY 12 (1) S984 - S985 1556-0864 2017/01 [Refereed][Not invited]
  • Nobuyuki Koyama, Yasutaka Watanabe, Yuki Iwai, Rumi Kawamura, Chihiro Miwa, Yoshiaki Nagai, Koichi Hagiwara, Shinichiro Koyama
    CHEMOTHERAPY 62 (3) 151 - 158 0009-3157 2017 [Refereed][Not invited]
    Background: Exon 19 deletion (Del19) and exon 21 L858R substitution (L858R), which account for 90% of epidermal growth factor receptor (EGFR) mutations as common mutations, are associated with favorable outcomes with EGFR-tyrosine kinase inhibitors (TKIs) compared with other uncommon EGFR mutations in non-small-cell lung cancer (NSCLC). However, whether there are differences in overall survival (OS) between patients with these common EGFR mutations remains controversial. Methods: The subjects studied were 74 NSCLC patients with common EGFR mutations treated with gefitinib or erlotinib. Using univariate and multivariate analyses, we retrospectively compared the clinicopahological characteristics of patients harboring Del19 with those harboring L858R. Results: Compared with patients harboring L858R, EGFR-TKIs provided a significant OS benefit in patients harboring Del19 (p = 0.024), as well as favorable therapeutic responses (p = 0.045) and progression-free survival (PFS) benefits (p = 0.031). In multivariate analyses, Del19 was independently associated with PFS (p = 0.029) and OS (p = 0.009), whereas no parameters other than pleural dissemination at the initial treatment were associated with EGFR mutation types. Conclusion: Del19 and L858R have distinct prognostic implications and may require individual therapeutic strategies. (C) 2017 S. Karger AG, Basel
  • Martin Reck, Koichi Hagiwara, Baohui Han, Sergei Tjulandin, Christian Grohe, Takashi Yokoi, Alessandro Morabito, Silvia Novello, Edurne Arriola, Olivier Molinier, Rose McCormack, Marianne Ratcliffe, Nicola Normanno
    JOURNAL OF THORACIC ONCOLOGY 11 (10) 1682 - 1689 1556-0864 2016/10 [Refereed][Not invited]
    Introduction: To offer patients with EGFR mutation positive advanced NSCLC appropriate EGFR tyrosine kinase inhibitor treatment, mutation testing of tumor samples is required. However, tissue/cytologic samples are not always available or evaluable. The large, noninterventional diagnostic ASSESS study (NCT01785888) evaluated the utility of circulating free tumor-derived DNA (ctDNA) from plasma for EGFR mutation testing. Methods: ASSESS was conducted in 56 centers (in Europe and Japan). Eligible patients (with newly diagnosed locally advanced/metastatic treatment-naive advanced NSCLC) provided diagnostic tissue/cytologic and plasma samples. DNA extracted from tissue/cytologic samples was subjected to EGFR mutation testing using local practices; designated laboratories performed DNA extraction/mutation testing of blood samples. The primary end point was level of concordance of EGFR mutation status between matched tissue/cytologic and plasma samples. Results: Of 1311 patients enrolled, 1288 were eligible. Concordance of mutation status in 1162 matched samples was 89% (sensitivity 46%, specificity 97%, positive predictive value 78%, and negative predictive value 90%). A group of 25 patients with apparent false-positive plasma results was overrepresented for cytologic samples, use of less sensitive tissue testing methodologies, and smoking habits associated with high EGFR mutation frequency, indicative of false negative tumor results. In cases in which plasma and tumor samples were tested with identical highly sensitive methods, positive predictive value/sensitivity were generally improved. Conclusions: These real-world data suggest that ctDNA is a feasible sample for EGFR mutation analysis. It is important to conduct mutation testing of both tumor and plasma samples in specialized laboratories, using robust/sensitive methods to ensure that patients receive appropriate treatments that target the molecular features of their disease. (C) 2016 International Association for the Study of Lung Cancer. Published by Elsevier Inc.
  • Yoshikazu Inoue, Koh Nakata, Etsuro Yamaguchi, Toru Arai, Chikatoshi Sugimoto, Yasuhiro Setoguchi, Toshio Ichiwata, Masahito Ebina, Kazutoshi Cho, Ryushi Tazawa, Haruyuki Ishii, Takahiro Kasai, Masanori Akira, Kanji Uchida, Hiroshi Kida, Sakae Homma, Koichiro Tatsumi, Arata Azuma, Koichi Hagiwara, Keisuke Tomii, Masanori Kitaichi, Masaru Suzuki, Kohnosuke Morimoto, Toshinori Takada, Hideaki Nakayama, Shinya Ohkouchi, Takahiro Tanaka, Masaki Hirose, Akiko Matsumuro
    EUROPEAN RESPIRATORY JOURNAL 48 0903-1936 2016/09 [Refereed][Not invited]
  • Kazuhiro Usui, Shunichi Sugawara, Masaru Nishitsuji, Yuka Fujita, Akira Inoue, Atsuto Mouri, Hiroshi Watanabe, Hiroshi Sakai, Ichiro Kinoshita, Yoshihito Ohhara, Makoto Maemondo, Hiroshi Kagamu, Koichi Hagiwara, Kunihiko Kobayashi
    LUNG CANCER 99 131 - 136 0169-5002 2016/09 [Refereed][Not invited]
    Background: Vascular endothelial growth factor (VEGF) plays a pivotal role in the pathogenesis of malignant pleural effusion (MPE). Here, a multicenter phase II trial to evaluate bevacizumab in non-squamous non-small cell lung carcinoma patients with MPE was conducted. Methods: Patients having MPE with no prior treatment and performance status of 0-2 received carboplatin (area under the curve: AUC 6; up to 6 cycles) and pemetrexed (500 mg/m(2)) with bevacizumab (15 mg/kg) every 3 weeks. The primary endpoint was the control rate of MPE without pleurodesis at 8 weeks after treatment. VEGF levels in plasma and MPE were measured by enzyme immunoassay. Results: Of 30 patients entered (median 66 years; 24 males; adenocarcinoma; 4 epidermal growth factor receptor: EGFR mutations), 28 patients (2 withdrawn patients) were given a median of 4 cycles of carboplatin, and 68% of the patients received maintenance pemetrexed with bevacizumab (median 8 cycles). At eight weeks, MPE was controlled without pleurodesis in 93% of treated patients (95% confidence interval: 77-99%). At the median follow-up time of 12.8 months, 78.6% of the cases required no pleurodesis. Response rate was 46%, and median progression-free survival (PFS) and overall survival (OS) were 8.2 months and 18.6 months, respectively. Toxicities of grade >= 3 included neutropenia (28.6%), thrombocytopenia (28.6%), proteinuria (3.6%), and hypertension (3.6%). Assessment of VEGF levels before treatment indicated that patients with low VEGF (<1000 pg/ml) in MPE frequently needed pleurodesis (p = 0.011), and that high VEGF (>= 100 pg/ml) in plasma was indicative of poor prognosis in the context of PFS (p = 0.012). Conclusion: The combination of bevacizumab with carboplatin and pemetrexed demonstrated efficacy with acceptable toxicities in patients with MPE. (C) 2016 Elsevier Ireland Ltd. All rights reserved.
  • Eisaku Miyauchi, Kazuhiro Usui, Shunichi Sugawara, Masaru Nishitsuji, Yuka Fujita, Atsuto Mouri, Hiroshi Watanabe, Hiroshi Sakai, Ichiro Kinoshita, Koichi Hagiwara
    ANNALS OF ONCOLOGY 27 0923-7534 2016/07 [Refereed][Not invited]
  • Takashi Hirama, Ayako Shiono, Hiroshi Egashira, Etsuko Kishi, Koichi Hagiwara, Hidetoshi Nakamura, Minoru Kanazawa, Makoto Nagata
    JOURNAL OF CLINICAL MICROBIOLOGY 54 (3) 699 - 704 0095-1137 2016/03 [Refereed][Not invited]
    The nontuberculous mycobacteria (NTM) cause miscellaneous disorders in humans, especially in the lungs, which present with a variety of radiological features. To date, knowledge of the pathogenic role of the Mycobacterium avium-intracellulare complex (MAC) in the human lung and the definitive criteria for initiating multidrug therapy are still lacking. However, there is little doubt that clarithromycin is the most efficacious drug among the various treatment regimens for lung NTM. In this study, with the use of a bridged nucleic acid (BNA) probe a detection system based on a real-time PCR (BNA-PCR) for the identification of the point mutations at position 2058 or 2059 in domain V of the 23S rRNA gene responsible for clarithromycin resistance was developed and has been assessed using MAC isolates from clinical samples. Out of 199 respiratory specimens, the drug susceptibility test demonstrated 12 strains resistant to clarithromycin, while the BNA-PCR showed 8 strains carrying the point mutation at position 2058 or 2059 of the 23S rRNA gene. This system revealed that there were mycobacterial strains resistant to clarithromycin which do not carry previously identified resistance genes. This paper documents a novel system for detecting clarithromycin-resistant strains and demonstrates that although these mutations are tacitly assumed to account for >90% of the reported resistant mutants, there is a significant fraction of resistant mutants that do not harbor these mutations. Therefore, unknown mechanisms affecting clarithromycin resistance remain to be elucidated.
  • Nicola Normanno, Koichi Hagiwara, Baohui Han, Sergei Tjulandin, Christian Grohe, Takashi Yokoi, Alessandro Morabito, Silvia Novello, Edurne Arriola, Olivier Molinier, Rose Mccormack, Marianne Ratcliffe, Martin Reck
    JOURNAL OF THORACIC ONCOLOGY 10 (9) S262 - S263 1556-0864 2015/09 [Refereed][Not invited]
  • Sergei Tjulandin, Baohui Han, Koichi Hagiwara, Nicola Normanno, Laksmi Wulandari, Konstantin Laktionov, Achmad Hudoyo, Yong He, Yi P. Zhang, Meng-Zhao Wang, Chien Ying Liu, Marianne Ratcliffe, Rose Mccormack, Martin Reck
    JOURNAL OF THORACIC ONCOLOGY 10 (9) S466 - S467 1556-0864 2015/09 [Refereed][Not invited]
  • Hagiwara K
    Nihon Naika Gakkai zasshi. The Journal of the Japanese Society of Internal Medicine 104 (9) 1808 - 1811 0021-5384 2015/09 [Refereed][Not invited]
  • Eisaku Miyauchi, Akira Inoue, Kunihiko Kobayashi, Makoto Maemondo, Shunichi Sugawara, Satoshi Oizumi, Hiroshi Isobe, Akihiko Gemma, Yasuo Saijo, Hirohisa Yoshizawa, Koichi Hagiwara, Toshihiro Nukiwa
    JAPANESE JOURNAL OF CLINICAL ONCOLOGY 45 (7) 670 - 676 0368-2811 2015/07 [Refereed][Not invited]
    Objective: Epidermal growth factor receptor tyrosine kinase inhibitors are effective as first-line therapy for advanced non-small cell lung cancer patients harboring epidermal growth factor receptor mutations. However, it is unknown whether second-line platinum-based chemotherapy after epidermal growth factor receptor tyrosine kinase inhibitor therapy could lead to better outcomes. We evaluated the efficacy of second-line platinum-based chemotherapy after gefitinib for advanced non-small cell lung cancers harboring epidermal growth factor receptormutations (the NEJ002 study). Methods: Seventy-one non-small cell lung cancers, treated with gefitinib as first-line therapy and then receiving platinum-based chemotherapy as second-line therapy were evaluated in NEJ002. Patients were evaluated for antitumor response to second-line chemotherapy by computed tomography according to the criteria of the Response Evaluation Criteria in Solid Tumors group (version 1.0). Results: Of the 71 patients receiving platinum-based chemotherapy after first-line gefitinib, a partial response was documented in 25.4% (18/71), stable disease in 43.7% (31/71) and progression of disease in 21.1% (15/71). The objective response and disease control rates were 25.4% (18/71) and 69% (49/71), respectively. There was no significant difference between first-and second-line chemotherapy in objective response and disease control rates for advanced non-small cell lung cancer harboring activating epidermal growth factor receptor mutations. In the analysis of epidermal growth factor receptor mutation types, the objective responses of deletions in exon 19 and a point mutation in exon 21 (L858R) were 27.3% (9/33) and 28.1% (9/32), respectively, but these differences between objective response rates were not significant. Conclusions: The efficacy of second-line platinum-based chemotherapy followed at progression by gefitinib was similar to first-line platinum-based chemotherapy, and epidermal growth factor receptor mutation types did not influence the efficacy of second-line platinum-based chemotherapy.
  • Tatsuro Fukuhara, Makoto Maemondo, Akira Inoue, Kunihiko Kobayashi, Shunichi Sugawara, Satoshi Oizumi, Hiroshi Isobe, Akihiko Gemma, Masao Harada, Hirohisa Yoshizawa, Ichiro Kinoshita, Yuka Fujita, Yasuo Saijo, Koichi Hagiwara, Satoshi Morita, Toshihiro Nukiwa
    LUNG CANCER 88 (2) 181 - 186 0169-5002 2015/05 [Refereed][Not invited]
    Introduction: Epidermal growth factor receptor (EGFR)-tyrosine kinase inhibitor (TKI) treatment is the standard therapy for non-small cell lung cancer (NSCLC) harbouring EGFR-activating mutations. The NEJ002 phase 3 clinical trial demonstrated the efficacy of EGFR-TKI; gefitinib was significantly superior in both progression-free survival (PFS) and objective response rate (ORR) than carboplatin plus paclitaxel. However, several cases showed no response. In this study, we performed further analysis of the characteristics of these non-responders. Methods: Available data from NEJ002 on maximum changes in tumour size were obtained from 103 cases (90.4%) and 110 cases (96.5%) in the carboplatin paclitaxel and gefitinib groups, respectively. Waterfall plots of maximum tumour size changes were created for non-responders. Results: Five (4.9%) and 9 (8.2%) cases in the carboplatin paclitaxel and gefitinib groups were non-responders, respectively. The mean pack years of the non-responders in the carboplatin paclitaxel and gefitinib groups were 0.33 and 31.7, respectively. The ORR of total smokers (61.5%) and heavy smokers (over 40 pack years, 52.6%) in the gefitinib group were significantly lower compared to people who have never smoked (80.0%) (P = 0.044 and P=0.020, respectively). Smoker cases also showed a tendency towards lower PFS and overall survival (05). In addition, the EGFR common mutation types did not affect PFS and OS in gefitinib-treated cases in NEJ002. However, in this study, the ORR and waterfall plots showed that gefitinib-treated non-responders who had a deletion in exon 19 in the EGFR gene exhibited a tendency towards a higher response compared to those with a L858R mutation. Conclusions: NSCLC patients with a smoking history or the EGFR L858R mutation may demonstrate a poorer response to gefitinib treatment. 0 2015 The Authors. Published by Elsevier Ireland Ltd.
  • Takashi Hirama, Hajime Mogi, Hitoshi Egashira, Eiji Yamamoto, Shigenari Kukisaki, Koichi Hagiwara, Osamu Takei
    CLINICA CHIMICA ACTA 445 122 - 126 0009-8981 2015/05 [Refereed][Not invited]
    Currently molecular techniques are a broadly accepted tool for diagnosis and are able to benefit patients in clinical practice. The polymerase chain reaction (PCR) has been especially incorporated into practical applications that are already in widespread use across the globe. With regard to the initial DNA extraction from clinically relevant samples, a number of commercially available kits are commonly used and are also designed to be easy to handle and less labor-intensive. In this study, the pressure system extracting DNA in column-based kit was developed, and its utility was compared with the centrifuge method using sputum from patients who were diagnosed with pneumonia. Also, due to the compact size and rapid processing time, the practical application of the pressure-based system incorporated into an automated pipetting machine was evaluated through clinical study. Our data suggests that DNA extraction by pressure was capable of serving as a substitute for the centrifuge method, and the compact and automatic nature of the pressure system device provided rapid and valuable information for clinical practice. (C) 2015 Elsevier B.V. All rights reserved.
  • Yasushi Yatabe, Keith M. Kerr, Ahmad Utomo, Pathmanathan Rajadurai, Van Khanh Tran, Xiang Du, Teh-Ying Chou, Ma. Luisa D. Enriquez, Geon Kook Lee, Jabed Iqbal, Shanop Shuangshoti, Jin-Haeng Chung, Koichi Hagiwara, Zhiyong Liang, Nicola Normanno, Keunchil Park, Shinichi Toyooka, Chun-Ming Tsai, Paul Waring, Li Zhang, Rose McCormack, Marianne Ratcliffe, Yohji Itoh, Masatoshi Sugeno, Tony Mok
    JOURNAL OF THORACIC ONCOLOGY 10 (3) 438 - 445 1556-0864 2015/03 [Refereed][Not invited]
    Introduction: The efficacy of epidermal growth factor receptor (EGFR) tyrosine kinase inhibitors in EGFR mutation-positive non-small-cell lung cancer (NSCLC) patients necessitates accurate, timely testing. Although EGFR mutation testing has been adopted by many laboratories in Asia, data are lacking on the proportion of NSCLC patients tested in each country, and the most commonly used testing methods. Methods: A retrospective survey of records from NSCLC patients tested for EGFR mutations during 2011 was conducted in 11 Asian Pacific countries at 40 sites that routinely performed EGFR mutation testing during that period. Patient records were used to complete an online questionnaire at each site. Results: Of the 22,193 NSCLC patient records surveyed, 31.8% (95% confidence interval: 31.2%-32.5%) were tested for EGFR mutations. The rate of EGFR mutation positivity was 39.6% among the 10,687 cases tested. The majority of samples were biopsy and/or cytology samples (71.4%). DNA sequencing was the most commonly used testing method accounting for 40% and 32.5% of tissue and cytology samples, respectively. A pathology report was available only to 60.0% of the sites, and 47.5% were not members of a Quality Assurance Scheme. Conclusions: In 2011, EGFR mutation testing practices varied widely across Asia. These data provide a reference platform from which to improve the molecular diagnosis of NSCLC, and EGFR mutation testing in particular, in Asia.
  • Naoto Morikawa, Yuji Minegishi, Akira Inoue, Makoto Maemondo, Kunihiko Kobayashi, Shunichi Sugawara, Masao Harada, Koichi Hagiwara, Shoji Okinaga, Satoshi Oizumi, Toshihiro Nukiwa, Akihiko Gemma
    EXPERT OPINION ON PHARMACOTHERAPY 16 (4) 465 - 472 1465-6566 2015/03 [Refereed][Not invited]
    Objective: To assess outcomes of elderly patients with advanced NSCLC harboring an EGFR mutation treated with gefitinib, as well as safety and impact on quality of life (QoL). Methods: We performed a retrospective analysis of pooled data from one Phase III and two Phase II studies of 71 patients aged >= 70 years with a performance status of 0-2. The main outcome measures were progression-free survival (PFS), overall survival (OS) and response rate (RR), as well as incidence of adverse events and time to 9.1% deterioration in QoL. Results: Median PFS (14.3 vs 5.7 months, p < 0.001) and overall RR (73.2 vs 26.5%, p < 0.001) in the gefitinib group were superior to those in the standard chemotherapy group, whereas median OS was not significantly different (30.8 vs 26.4 months, p = 0.42). Elevation of aspartate transaminase and/or alanine transaminase (18.3%) was the most common adverse event, and one treatment-related death (pneumonitis) occurred. Time to 9.1% deterioration in the QoL domains of pain and dyspnea, anxiety, and daily functioning was similar between the two age groups. Conclusion: First-line gefitinib is efficacious with acceptable toxicity in relatively fit elderly patients with advanced NSCLC harboring an EGFR mutation.
  • Shinsuke Hashida, Junichi Soh, Shinichi Toyooka, Tomoaki Tanaka, Masashi Furukawa, Kazuhiko Shien, Hiromasa Yamamoto, Hiroaki Asano, Kazunori Tsukuda, Koichi Hagiwara, Shinichiro Miyoshi
    ONCOLOGY REPORTS 32 (1) 145 - 152 1021-335X 2014/07 [Refereed][Not invited]
    The T790M mutation in the epidermal growth factor receptor (EGFR) gene is known to be associated with the acquired resistance of lung adenocarcinoma patients to EGFR-tyrosine kinase inhibitors (EGFR-TKIs). The minor T790M mutant allele is occasionally detected in EGFR-TKI-naive tumor samples, yet findings concerning the clinical impact of the minor T790M mutation vary among previous studies. In the present study, we assessed the clinical impact of the minor T790M mutation using a novel, highly sensitive assay combining high-resolution melting (HRM), mutant-enriched PCR and co-amplification at a lower denaturation temperature (COLD)-PCR. We determined the T790M mutational status in 146 surgically resected lung adenocarcinomas without a history of EGFR-TKI treatment using mutant-enriched COLD-HRM (MEC-HRM) and standard HRM assays. The sensitivities of the MEC-HRM and standard HRM assays for the detection of T790M-mutant alleles among wild-type alleles were 0.01 and 10%, respectively. Although the T790M mutation was not detected using a standard HRM assay, we identified 19 (13%) T790M mutations using the MEC-HRM assay and defined these 19 mutations as minor T790M mutations. The proportion of T790M alleles was <0.1% in 17 (84%) of the 19 samples. Multivariate analyses revealed that a minor T790M mutation was significantly associated with the presence of EGFR exon 19 deletions or the L858R mutation (both of which are drug-sensitive EGFR mutations) (P=0.04). In conclusion, the minor EGFR T790M mutations were present in 13% of EGFR-TKI-naive surgically resected lung adenocarcinomas and were associated with drug-sensitive EGFR mutations.
  • Satoshi Oizumi, Shunichi Sugawara, Koichi Minato, Toshiyuki Harada, Akira Inoue, Yuka Fujita, Makoto Maemondo, Hirohisa Yoshizawa, Kazuhiko Ito, Akihiko Gemma, Masaru Nishitsuji, Masao Harada, Hiroshi Isobe, Ichiro Kinoshita, Satoshi Morita, Kunihiko Kobayashi, Koichi Hagiwara, Minoru Kurihara, Toshihiro Nukiwa
    JOURNAL OF CLINICAL ONCOLOGY 32 (15) 0732-183X 2014/05 [Refereed][Not invited]
  • Koichi Hagiwara, Katsuya Ogata, Tsuyoshi Okamoto, Taira Uehara, Naruhito Hironaga, Hiroshi Shigeto, Jun-ichi Kira, Shozo Tobimatsu
    CLINICAL NEUROPHYSIOLOGY 125 (5) 1021 - 1029 1388-2457 2014/05 [Refereed][Not invited]
    Objective: Age-related changes are well documented in the primary somatosensory cortex (SI). Based on previous somatosensory evoked potential studies, the amplitude of N20 typically increases with age probably due to cortical disinhibition. However, less is known about age-related change in the secondary somatosensory cortex (SII). The current study quantified age-related changes across SI and SII mainly based on oscillatory activity indices measured with magnetoencephalography. Methods: We recorded somatosensory evoked magnetic fields (SEFs) to right median nerve stimulation in healthy young and old subjects and assessed major SEF components. Then, we evaluated the phase-locking factor (PLF) for local field synchrony on neural oscillations and the weighted phase-lag index (wPLI) for cortico-cortical synchrony between SI and SII. Results: PLF was significantly increased in SI along with the increased amplitude of N20m in the old subjects. PLF was also increased in SII associated with a shortened peak latency of SEFs. wPLI analysis revealed the increased coherent activity between SI and SII. Conclusions: Our results suggest that the functional coupling between SI and SII is influenced by the cortical disinhibition due to normal aging. Significance: We provide the first electrophysiological evidence for age-related changes in oscillatory neural activities across the somatosensory areas. (C) 2013 International Federation of Clinical Neurophysiology. Published by Elsevier Ireland Ltd. All rights reserved.
  • Takashi Hirama, Shohei Minezaki, Takefumi Yamaguchi, Etsuko Kishi, Keiji Kodama, Hiroshi Egashira, Kunihiko Kobayashi, Makoto Nagata, Toshiaki Ishii, Manabu Nemoto, Masahiko Tanaka, Koichi Fukunaga, Minoru Kanazawa, Koichi Hagiwara
    Respiratory medicine 108 (2) 395 - 404 0954-6111 2014/02 [Refereed][Not invited]
    UNLABELLED: Identification of the causative pathogen(s) of pneumonia would allow the selection of effective antibiotics and thus reduce the mortality rate and the emergence of drug-resistant pathogens. To identify such pathogens and to obtain these benefits, it is necessary that a clinical test is rapid, accurate, easily performed, and cost-effective. Here, we devised a PCR-based test, named HIRA-TAN, which is able to discriminate therapeutic targets from commensal organisms (e.g. Streptococcus pneumoniae or Haemophilus influenzae) and to detect foreign organisms (e.g. Mycoplasma pneumoniae or Legionella pneumophila) in the sputum. The utility of this system was validated in a prospective study, using sputum samples from patients with pneumonia. 568 patients were enrolled and the HIRA-TAN assay identified the causative pathogens with an accuracy of 96.7% for H. influenzae; 93.2% for Pseudomonas aeruginosa; 80.6% for Klebsiella pneumoniae; 90.9% for Moraxella catarrhalis; 87.5% for Escherichia coli; 78.1% for MRSA and 91.6% for S. pneumoniae. Overall the HIRA-TAN procedure was able to identify the causative pathogens of pneumonia in 60% of the cases. Additionally, this procedure was able to determine when the pneumonia-causing organism was a commensal organism or a foreign organism in a single assay. The HIRA-TAN approach yielded reproducible results and provided valuable information to plan the course of treatment of pneumonia. Through the rapid identification of the causative pathogens, the HIRA-TAN will promote targeted treatments for pneumonias. CLINICAL TRIALS REGISTRATION: UMIN000001694.
  • Satoshi Watanabe, Yuji Minegishi, Hirohisa Yoshizawa, Makoto Maemondo, Akira Inoue, Shunichi Sugawara, Hiroshi Isobe, Masao Harada, Yoshiki Ishii, Akihiko Gemma, Koichi Hagiwara, Kunihiko Kobayashi
    JOURNAL OF THORACIC ONCOLOGY 9 (2) 189 - 194 1556-0864 2014/02 [Refereed][Not invited]
    Introduction: In non-small-cell lung cancer, an exon 19 deletion and an L858R point mutation in the epidermal growth factor receptor (EGFR) are predictors of a response to EGFR-tyrosine kinase inhibitors. However, it is uncertain whether other uncommon EGFR mutations are associated with sensitivity to EGFR-tyrosine kinase inhibitors. Methods: A post-hoc analysis to assess prognostic factors was performed with the use of patients with EGFR mutations (exon 19 deletion, L858R, G719X, and L861Q) who were treated with gefitinib in the NEJ002 study, which compared gefitinib with carboplatin-paclitaxel as the first-line therapy. Results: In the NEJ002 study, 225 patients with EGFR mutations received gefitinib at any treatment line. The Cox proportional hazards model indicated that performance status, response to chemotherapy, response to gefitinib, and mutation types were significant prognostic factors. Overall survival (OS) was significantly shorter among patients with uncommon EGFR mutations (G719X or L861Q) compared with OS of those with common EGFR mutations (12 versus 28.4 months; p = 0.002). In the gefitinib group (n = 114), patients with uncommon EGFR mutations had a significantly shorter OS (11.9 versus 29.3 months; p < 0.001). By contrast, OS was similar between patients with uncommon mutations and those with common mutations in the carboplatin-paclitaxel group (n = 111; 22.8 versus 28 months; p = 0.358). Conclusions: The post-hoc analyses clearly demonstrated shorter survival for gefitinib-treated patients with uncommon EGFR mutations compared with the survival of those with common mutations and suggest that the first-line chemotherapy may be relatively effective for non-small-cell lung cancer with uncommon EGFR mutations.
  • Rinako Ishikawa, Tetsuya Okano, Tomohiko Mio, Ai Masumoto, Akiko Kawashima, Yuri Maeno, Susumu Yamazaki, Hisayoshi Daito, Ou Yamaguchi, Yoshitake Murayama, Koichi Hagiwara, Kunihiko Kobayashi
    RESPIROLOGY 18 27 - 27 1323-7799 2013/11 [Refereed][Not invited]
  • Yotaro Takaku, Kazuyuki Nakagome, Takehito Kobayashi, Tomoyuki Soma, Koichi Hagiwara, Minoru Kanazawa, Makoto Nagata
    RESPIROLOGY 18 114 - 114 1323-7799 2013/11 [Refereed][Not invited]
  • Baohui Han, Sergei Tjulandin, Nicola Normanno, Koichi Hagiwara, Martin Reck, Gael Mcwalter, Rose Mccormack
    JOURNAL OF THORACIC ONCOLOGY 8 S1098 - S1099 1556-0864 2013/11 [Refereed][Not invited]
  • Martin Reck, Koichi Hagiwara, Sergei Tjulandin, Baohui Han, Gael Mcwalter, Rose Mccormack, Nicola Normanno
    JOURNAL OF THORACIC ONCOLOGY 8 S483 - S484 1556-0864 2013/11 [Refereed][Not invited]
  • Hiroki Kabata, Toru Satoh, Masaharu Kataoka, Yuichi Tamura, Tomohiko Ono, Miyuki Yamamoto, Huqun, Koichi Hagiwara, Keiichi Fukuda, Tomoko Betsuyaku, Koichiro Asano
    RESPIROLOGY 18 (7) 1076 - 1082 1323-7799 2013/10 [Refereed][Not invited]
    Background and objectiveMutation of bone morphogenetic protein receptor type 2 (BMPR2) is a cause of pulmonary arterial hypertension (PAH). We measured the prevalence of this mutation and its impact on the phenotypes and long-term clinical outcomes in Japanese patients. MethodsBetween 1999 and 2007, we consecutively enrolled and, until March 2012, followed 49 Japanese patients with PAH, including nine familial cases from seven families. We genotyped BMPR2, using direct sequencing and multiplex ligation-dependent probe amplification, to examine (i) the prevalence of BMPR2 mutations and gene rearrangement, (ii) the relationship between BMPR2 genotype and clinical phenotypes, and (iii) the long-term clinical outcomes of mutation carriers versus non-carriers under state-of-the-art medical therapy. ResultsBMPR2 mutations were present in four of the seven families (57%) and in 14 of the 40 patients (35%) with sporadic PAH. The mean age at onset of PAH was 37.4 years in BMPR2 carriers, versus 25.9 years in non-carriers (P=0.0025). The gender distribution and hemodynamic status at time of diagnosis were similar regardless of the mutation status. The 5-year survival rate after diagnosis of PAH was 88.5% in BMPR2 mutation carriers versus 80.9% in non-carriers (ns). ConclusionsThe prevalence of BMPR2 mutations in Japanese with PAH was similar to that reported in other populations. At onset of PAH, BMPR2 mutation non-carriers were, on average, younger than carriers, possibly due to the heterogeneity of this subpopulation. With state-of-the-art therapy, the long-term survival of patients with PAH was high, regardless of the mutation status.
  • Masumoto A, Hirama T, Kawana H, Kawashima A, Shiono A, Mio T, Nagata M, Hagiwara K, Kanazawa M
    Nihon Naika Gakkai zasshi. The Journal of the Japanese Society of Internal Medicine 102 (10) 2679 - 2681 0021-5384 2013/10 [Refereed][Not invited]
  • Shohei Minezaki, Hidetoshi Nakamura, Takashi Hirama, Ayako Shiono, Akiko Kawashima, Makoto Nagata, Koichi Hagiwara, Minoru Kanazawa
    EUROPEAN RESPIRATORY JOURNAL 42 0903-1936 2013/09 [Refereed][Not invited]
  • Tomoko Suzuki, Masaya Suzuki, Gregory Downey, Koichi Hagiwara
    EUROPEAN RESPIRATORY JOURNAL 42 0903-1936 2013/09 [Refereed][Not invited]
  • Akihiko Miyanaga, Kumi Shimizu, Rintaro Noro, Masahiro Seike, Kazuhiro Kitamura, Seiji Kosaihira, Yuji Minegishi, Takehito Shukuya, Akinobu Yoshimura, Masashi Kawamoto, Shinichi Tsuchiya, Koichi Hagiwara, Manabu Soda, Kengo Takeuchi, Nobuyuki Yamamoto, Hiroyuki Mano, Yuichi Ishikawa, Akihiko Gemma
    BMC CANCER 13 262  1471-2407 2013/05 [Refereed][Not invited]
    Background: The EML4-ALK (echinoderm microtubule-associated protein-like 4 gene and the anaplastic lymphoma kinase gene) fusion oncogene represents a novel molecular target in a small subset of non-small-cell lung cancers (NSCLCs). The EML4-ALK fusion gene occurs generally in NSCLC without mutations in epidermal growth factor receptor (EGFR) and KRAS. Case presentation: We report that a case of EML4-ALK-positive NSCLC with EGFR mutation had a response of stable disease to both an EGFR tyrosine kinase inhibitor (EGFR-TKI) and ALK inhibitor. Conclusions: We described the first clinical report of a patient with EML4-ALK-positive NSCLC with EGFR mutation that had a response of stable disease to both single-agent EGFR-TKI and ALK inhibitor. EML4-ALK translocation may be associated with resistance to EGFR-TKI, and EGFR signaling may contribute to resistance to ALK inhibitor in EML4-ALK-positive NSCLC.
  • Koichi Hagiwara, Kunihiko Kobayashi
    CANCER SCIENCE 104 (3) 291 - 297 1347-9032 2013/03 [Refereed][Not invited]
    Mutations in the epidermal growth factor receptor (EGFR) gene confer it with cancer driver gene functions in non-small cell lung cancer (NSCLC). Epidermal growth factor receptor -tyrosine kinase inhibitors are effective agents against NSCLC with a mutated EGFR gene. Accordingly, many guidelines recommend the use of an EGFR mutation test in NSCLC. However, not all patients are tested in most countries where tissue samples are mainly used for the test. As of 2011, most of the patients with advanced NSCLC are tested in Japan, and the use of cytological samples has significantly contributed to this success. A portion of samples used to determine a definite diagnosis of NSCLC, either tissue samples or cytological samples, is ensured to contain cancer cells, and is then investigated by an EGFR mutation test that is applicable to both tissue samples and cytological samples. Cytological samples now account for one-third of all the samples investigated. EGFR mutation is detected in cytological samples at a similar rate with tissue samples. The criterion ensuring an EGFR mutation test to have satisfactory sensitivity and specificity for use in both tissue and cytological samples is presented. Cytological samples are valuable clinical sources being collected less invasively than tissue samples, and should therefore be extensively used in EGFR mutation testing.
  • Kunihiko Kobayashi, Koichi Hagiwara
    TARGETED ONCOLOGY 8 (1) 27 - 33 1776-2596 2013/03 [Refereed][Not invited]
    Before 2009, nonsmall cell lung cancer (NSCLC) was one disease entity treated by cytotoxic chemotherapy that provided a response rate of 20-35 % and a median survival time (MST) of 10-12 months. In 2004, it was found that activated mutations of the epidermal growth factor receptor (EGFR) gene were present in a subset of NSCLC and that tumors with EGFR mutations were highly sensitive to EGFR tyrosine kinase inhibitors (TKI). Four phase III studies (North East Japan (NEJ) 002, West Japan Thoracic Oncology Group (WJTOG) 3405, OPTIMAL, and EUROTAC) prospectively compared TKI (gefitinib or erlotinib) with cytotoxic chemotherapy as first-line therapy in EGFR-mutated NSCLC. These studies confirmed that progression-free survival (PFS) with TKIs (as the primary endpoint) was significantly longer than that with standard chemotherapy (hazard ratio [HR] = 0.16-0.49) from 2009 to 2011. Although the NEJ 002 study showed identical overall survival (OS) between the arms (HR = 0.89), quality of life (QoL) was maintained much longer in patients treated with gefitinib. In conclusion, TKI should be considered as the standard first-line therapy in advanced EGFR-mutated NSCLC. Since 2009, a new step has been introduced in the treatment algorithm for advanced NSCLC.
  • Seung Tae Kim, Hae-Yun Jung, Jae Sook Sung, Uk Hyun Jo, Tomoaki Tanaka, Koichi Hagiwara, Kyong Hwa Park, Sang Won Shin, Jun Suk Kim, Yeul Hong Kim
    AMERICAN JOURNAL OF CLINICAL ONCOLOGY-CANCER CLINICAL TRIALS 36 (1) 57 - 63 0277-3732 2013/02 [Refereed][Not invited]
    Background: Epidermal growth factor receptor (EGFR) mutations as prognostic or predictive marker in patients with non-small cell lung cancer (NSCLC) have been used widely. However, it may be difficult to get tumor tissue for analyzing the status of EGFR mutation status in large proportion of patients with advanced disease. Patients and Methods: We obtained pairs of tumor and serum samples from 57 patients with advanced NSCLC, between March 2006 and January 2009. EGFR mutation status from tumor samples was analyzed by genomic polymerase chain reaction and direct sequence and EGFR mutation status from serum samples was determined by the peptide nucleic acid locked nucleic acid polymerase chain reaction clamp. Results: EGFR mutations were detected in the serum samples of 11 patients and in the tumor samples of 12 patients. EGFR mutation status in the serum and tumor samples was consistent in 50 of the 57 pairs (87.7%). There was a high correlation between the mutations detected in serum sample and the mutations detected in the matched tumor sample (correlation index 0.62; P < 0.001). Twenty-two of 57 patients (38.5%) received EGFR-tyrosine kinase inhibitors as any line therapy. The response for EGFR- tyrosine kinase inhibitors was significantly associated with EGFR mutations in both tumor samples and serum samples (P < 0.05). There was no significant differences in overall survival according to the status of EGFR mutations in both serum and tumor samples (P > 0.05). Conclusions: Serum sample might be alternatively used in the difficult time of getting tumor tissue for analyzing the status of EGFR mutation status in patients with advanced NSCLC.
  • Yutaka Usui, Akiko Kaga, Fumikazu Sakai, Ayako Shiono, Ken-ichiro Komiyama, Koichi Hagiwara, Minoru Kanazawa
    BMJ OPEN 3 (7) 2044-6055 2013 [Refereed][Not invited]
    Objectives To assess clinical, laboratory and radiographic findings associated with outcomes and to clarify more practical ways to predict hospital mortality in patients with acute exacerbation (AE) of chronic fibrosing interstitial pneumonia (CFIP). Design Single-centre retrospective cohort study. Setting University Hospital in Japan. Participants We identified 51 consecutive patients with AE of idiopathic CFIP through multidisciplinary discussion. Patients who had connective tissue disease, drug-induced lung disease, pneumoconiosis, hypersensitivity pneumonitis, sarcoidosis, pulmonary histiocytosis, lymphangioleiomyomatosis and eosinophilic pneumonia were excluded. Interventions There were no interventions. Main outcome measures The main outcome was determination of in-hospital mortality predictors. Other outcomes included clinical, laboratory and radiographic differences between non-survivors and survivors in patients with AE of CFIP. Results The mean age of the patients with AE of CFIP was 71years. Compared with survivors, non-survivors had a significantly shorter duration of symptoms before admission, lower prevalence of peripheral distribution of ground-glass opacity and centrilobular emphysema (CLE) on thin-section CT, lower peripheral lymphocyte count, higher brain natriuretic peptide titre, lower Pao(2):Fio(2) (P:F) ratio, higher prevalence of systemic inflammatory response syndrome (SIRS) and higher SIRS score on admission (p=0.0069, 0.0032, 0.015, 0.040, 0.0098, 0.012, 9.9x10(-7) and 5.4x10(-6), respectively). Multivariate analysis revealed SIRS (HR=6.2810, p=0.015), CLE (HR=0.0606, p=3.6x10(-5)) and serum procalcitonin level (HR=2.7110, p=0.022) to be independent predictors of in-hospital mortality. A Kaplan-Meier estimate on the basis of stratification according to the presence or absence of SIRS and CLE demonstrated a distinct survival curve for each subset of patients. Conclusions Distinct survival curves documented by stratification according to the presence or absence of SIRS and CLE may provide basic information for a rational management strategy for patients with AE of CFIP on admission.
  • Yotaro Takaku, Tomoyuki Soma, Fuyumi Nishihara, Kazuyuki Nakagome, Takehito Kobayashi, Koichi Hagiwara, Minoru Kanazawa, Makoto Nagata
    INTERNATIONAL ARCHIVES OF ALLERGY AND IMMUNOLOGY 161 107 - 117 1018-2438 2013 [Refereed][Not invited]
    Background: Omalizumab, an anti-immunoglobulin E monoclonal antibody, has shown an inhibitory effect on airway inflammation, which may be associated with clinical improvement of severe asthma. This study evaluated changes in airway inflammation and cytokine release by the peripheral blood mononuclear cells (PBMCs) of Japanese patients with severe asthma after administration of omalizumab. Methods: Sixteen Japanese patients with severe asthma who were allergic to house-dust mites were enrolled in this study. Eight received omalizumab every 2 or 4 weeks for 16 weeks, and 8 control subjects were treated with conventional drug treatment. Changes in clinical scores for sputum eosinophils and levels of fraction of exhaled nitric oxide (FeNO) were measured at the time of enrollment and at week 16. Cytokines from PBMCs stimulated by house-dust mite (Dermatophagoides farinae) or ionomycin/phorbol myristate acetate (PMA) were measured at baseline and at week 16. Results: In the omalizumab-treated group, decreases in sputum eosinophils and FeNO were observed following treatment. Furthermore, the ex vivo production of interleukin (IL)-5 by PBMCs in response to both mite allergen and ionomycin/PMA decreased significantly. In contrast, interferon (IFN)-gamma production was unchanged. There were no changes in any of the parameters observed in the control group. Conclusion: Omalizumab exerts inhibitory effects on airway inflammation in Japanese patients with severe allergic asthma. This treatment attenuates production of IL-5 by PBMCs stimulated with both a specific allergen and a nonspecific activator. Reduction of the Th2 inflammatory cascade likely contributes to clinical benefits; however, further studies are required to clarify these results due to the small sample size in this study. Copyright (C) 2013 S. Karger AG, Basel
  • Akiko Kawashima, Tomoko Suzuki, Fuyumi Nishihara, Takehito Kobayashi, Yotaro Takaku, Kazuyuki Nakagome, Tomoyuki Soma, Koichi Hagiwara, Minoru Kanazawa, Makoto Nagata
    INTERNATIONAL ARCHIVES OF ALLERGY AND IMMUNOLOGY 161 10 - 15 1018-2438 2013 [Refereed][Not invited]
    Background: Neutrophils are often increased in the airways of either chronic severe asthma or acute exacerbations. Neutrophils that have migrated in response to interleukin-8 (IL8) may lead eosinophils to accumulate in the airways of patients with asthma and possibly aggravate the disease. In this study, we investigated whether formoterol modified the trans-basement membrane migration (TBM) of eosinophils stimulated with neutrophils and IL-8. Methods: Neutrophils and eosinophils were isolated from peripheral blood obtained from healthy donors. Eosinophil TBM was examined using a modified Boyden's chamber technique. Neutrophils were preincubated with or without formoterol (0.1 mu M) at 37 degrees C for 30 min. Eosinophils were added to the upper compartment of a chamber with a Matrigel-coated transwell insert. Medium containing preincubated neutrophils and IL-8 was added to the lower compartment of the chamber. After a 90-minute incubation, the eosinophils that had migrated into the lower chamber were calculated using eosinophil peroxidase assays. Results: A combination of neutrophils and IL-8 significantly induced the eosinophil TBM; formoterol alone had no effect. However, formoterol modestly but significantly attenuated the TBM of eosinophils stimulated with neutrophils and IL-8. Conclusion: These results suggest that formoterol may act as a therapeutic agent on enhanced eosinophilic inflammation in acute exacerbation or persistent, severe asthma. The effect of formoterol likely involves the inhibition of neutrophil activation. Copyright (C) 2013 S. Karger AG, Basel
  • Hisayoshi Daito, Motoi Suzuki, Jun Shiihara, Paul E. Kilgore, Hitoshi Ohtomo, Konosuke Morimoto, Masayuki Ishida, Taro Kamigaki, Hitoshi Oshitani, Masahiro Hashizume, Wataru Endo, Koichi Hagiwara, Koya Ariyoshi, Shoji Okinaga
    Thorax 68 (6) 544 - 550 1468-3296 2013 [Refereed][Not invited]
    Background: On 11 March 2011, the Tohoku earthquake and tsunami struck off the coast of northeastern Japan. Within 3 weeks, an increased number of pneumonia admissions and deaths occurred in local hospitals. Methods: A multicentre survey was conducted at three hospitals in Kesennuma City (population 74 000), northern Miyagi Prefecture. All adults aged ≥18 years hospitalised between March 2010 and June 2011 with community-acquired pneumonia were identified using hospital databases and medical records. Segmented regression analyses were used to quantify changes in the incidence of pneumonia. Results: A total of 550 pneumonia hospitalisations were identified, including 325 during the pre-disaster period and 225 cases during the post-disaster period. The majority (90%) of the post-disaster pneumonia patients were aged ≥65 years, and only eight cases (3.6%) were associated with near-drowning in the tsunami waters. The clinical pattern and causative pathogens were almost identical among the pre-disaster and post-disaster pneumonia patients. A marked increase in the incidence of pneumonia was observed during the 3-month period following the disaster the weekly incidence rates of pneumonia hospitalisations and pneumonia-associated deaths increased by 5.7 times (95% CI 3.9 to 8.4) and 8.9 times (95% CI 4.4 to 17.8), respectively. The increases were largest among residents in nursing homes followed by those in evacuation shelters. Conclusions: A substantial increase in the pneumonia burden was observed among adults after the Tohoku earthquake and tsunami. Although the exact cause remains unresolved, multiple factors including population aging and stressful living conditions likely contributed to this pneumonia outbreak.
  • Kenichiro Komiyama, Ai Masumoto, Fuyumi Nishihara, Takehito Kobayashi, Tomoyuki Soma, Koichi Hagiwara, Minoru Kanazawa, Makoto Nagata
    Japanese Journal of Allergology 62 (12) 1631 - 1641 1347-7935 2013 [Refereed][Not invited]
    Background: ImmunoCAP® Rapid is a rapid test kit to measure the allergen-specific IgE to the eight major inhalation allergen (cat, mite, orchard grass, ragweed, wormwood, dog, cockroach, Japan cedar). Methods: We performed ImmunoCAP® Rapid 83 patients with allergic disease (26 males, 57 females, median aged 43 years, 53 of asthma, 43 of allergic rhinitis) in our allergy center. ImmunoCAP® Rapid results were compared with those of skin prick test (SPT). Results: Although total positive allergens of SPT were higher than that of ImmunoCAP® Rapid (26.5% vs 22.5%, p< O.05), there was no significantly difference of each positive allergen between two tests. The rate of ImmunoCAP® Rapid to Japan cedar was almost equivalent to SPT in all patients (68.7% vs 55.4%, p = 0.07). In contrast, the rate of ImmunoCAP® Rapid to Japan cedar was higher than SPT in patients with rhinitis (90.4% vs 71.4%, p< O.05). Efficiency between ImmunoCAP® Rapid and SPT was 86.4%, sensitivity was 66.9%, and specificity was 93.4%. The reactivity of ImmunoCAP® Rapid to allergens significantly correlated with sizes of SPT (erythema r = 0.645, urticaria: r = 0.657). Conclusion: Although identification rate in the screening ImmunoCAP® Rapid slightly inferior to SPT, this test system was useful for diagnosis of Japan cedar and mite.
  • Manabu Soda, Kazutoshi Isobe, Akira Inoue, Makoto Maemondo, Satoshi Oizumi, Yuka Fujita, Akihiko Gemma, Yoshihiro Yamashita, Toshihide Ueno, Kengo Takeuchi, Young Lim Choi, Hitoshi Miyazawa, Tomoaki Tanaka, Koichi Hagiwara, Hiroyuki Mano
    CLINICAL CANCER RESEARCH 18 (20) 5682 - 5689 1078-0432 2012/10 [Refereed][Not invited]
    Purpose: EML4-ALK is a lung cancer oncogene, and ALK inhibitors show marked therapeutic efficacy for tumors harboring this fusion gene. It remains unsettled, however, how the fusion gene should be detected in specimens other than formalin-fixed, paraffin-embedded tissue. We here tested whether reverse transcription PCR (RT-PCR)-based detection of EML4-ALK is a sensitive and reliable approach. Experimental Design: We developed a multiplex RT-PCR system to capture ALK fusion transcripts and applied this technique to our prospective, nationwide cohort of non-small cell lung cancer (NSCLC) in Japan. Results: During February to December 2009, we collected 916 specimens from 853 patients, quality filtering of which yielded 808 specimens of primary NSCLC from 754 individuals. Screening for EML4-ALK and KIF5B-ALK with our RT-PCR system identified EML4-ALK transcripts in 36 samples (4.46%) from 32 individuals (4.24%). The RT-PCR products were detected in specimens including bronchial washing fluid (n = 11), tumor biopsy (n = 8), resected tumor (n = 7), pleural effusion (n = 5), sputum (n = 4), and metastatic lymph node (n = 1). The results of RT-PCR were concordant with those of sensitive immunohistochemistry with ALK antibodies. Conclusions: Multiplex RT-PCR was confirmed to be a reliable technique for detection of ALK fusion transcripts. We propose that diagnostic tools for EML4-ALK should be selected in a manner dependent on the available specimen types. FISH and sensitive immunohistochemistry should be applied to formalin-fixed, paraffin-embedded tissue, but multiplex RT-PCR is appropriate for other specimen types. Clin Cancer Res; 18(20); 5682-9. (C) 2012 AACR.
  • Skronski Michal, Szpechcinski Adam, Jagus Paulina, Tanaka Tomoaki, Struniawski Radoslaw, Hagiwara Koichi, Chorostowska-Wynimko Joanna
    EUROPEAN RESPIRATORY JOURNAL 40 0903-1936 2012/09 [Refereed][Not invited]
  • Makoto Maemondo, Yuji Minegishi, Akira Inoue, Kunihiko Kobayashi, Masao Harada, Shoji Okinaga, Naoto Morikawa, Satoshi Oizumi, Tomoaki Tanaka, Hiroshi Isobe, Shoji Kudoh, Koichi Hagiwara, Toshihiro Nukiwa, Akihiko Gemma
    JOURNAL OF THORACIC ONCOLOGY 7 (9) 1417 - 1422 1556-0864 2012/09 [Refereed][Not invited]
    Introduction: Recent studies have demonstrated that first-line treatment with gefitinib, an epidermal growth factor receptor (EFGR)-targeted tyrosine kinase inhibitor, is significantly superior to standard chemotherapy for advanced non-small-cell lung cancer (NSCLC) harboring EGFR sensitive mutations. Meanwhile, the efficacy of gefitinib therapy among elderly populations diagnosed with EGFR-mutated NSCLC has not yet been elucidated. The purpose of this study was to investigate the efficacy and feasibility of gefitinib for chemotherapy-naive patients aged 75 or older with NSCLC harboring EGFR mutations; generally, these patients have no indication for treatment with platinum doublets. Methods: Chemotherapy-naive patients aged 75 years or older with performance status 0 to 1 and advanced NSCLC harboring EGFR mutations, as determined by the peptide nucleic acid-locked nucleic acid polymerase chain reaction clamp method, were enrolled. The enrolled patients received 250 mg/day of gefitinib orally. Results: Between January 2008 and May 2009, 31 patients were enrolled, all of whom were eligible. The median age was 80 (range, 75-87) years. Twenty-five patients (81%) were women, and 30 patients (97%) had adenocarcinoma. The overall response rate was 74% (95% confidence interval, 58%-91%), and the disease control rate was 90%. The median progression-free survival was 12.3 months. The common adverse events were rash, diarrhea, and liver dysfunction. One treatment-related death because of interstitial lung disease occurred. Conclusions: This is the first study that verified safety and efficacy of first-line treatment with gefitinib in elderly patients having advanced NSCLC with EGFR mutation. Considering its strong antitumor activity and mild toxicity, first-line gefitinib may be preferable to standard chemotherapy for this population.
  • Takeo Ishii, Koichi Hagiwara, Shinobu Ikeda, Tomio Arai, Makiko Naka Mieno, Toshio Kumasaka, Masaaki Muramatsu, Motoji Sawabe, Akihiko Gemma, Kozui Kida
    COPD-JOURNAL OF CHRONIC OBSTRUCTIVE PULMONARY DISEASE 9 (4) 409 - 416 1541-2555 2012/08 [Refereed][Not invited]
    Surfactant protein D (SFTPD) is a lung-specific anti-inflammatory factor that antagonizes inflammation by inhibiting oxidative stress and stimulating innate immunity. Variations in SFTPA2 and SFTPB, genes for other surfactant proteins, have been associated with lung cancer. We therefore investigated associations between SFTPD variations and lung cancer as well as emphysema and interstitial pneumonia, which are characterized by chronic inflammation from which lung cancer often arises. DNA from 1342 autopsy samples, including those from 140 subjects with lung cancer, was investigated. The single nucleotide polymorphism (SNP) rs721917, which results in methionine being exchanged for threonine at amino acid 11 (the Met11Thr variation), tended to be associated with emphysema and was associated with interstitial pneumonia and lung cancer. A haplotype analysis revealed that the haplotypes associated with emphysema and lung cancer differed from that associated with interstitial pneumonia, suggesting a differential role for SFTPD in the development of these diseases. A mediating analysis did not reveal a mediating effect exerted by emphysema or interstitial pneumonia on lung cancer. Our results suggested that SFTPD plays a role in the development of lung cancer and that the role for lung cancer may differ from that for interstitial pneumonia.
  • Hagiwara K
    Nihon rinsho. Japanese journal of clinical medicine 70 Suppl 6 521 - 525 0047-1852 2012/08 [Refereed][Not invited]
  • Satoshi Oizumi, Kunihiko Kobayashi, Akira Inoue, Makoto Maemondo, Shunichi Sugawara, Hirohisa Yoshizawa, Hiroshi Isobe, Masao Harada, Ichiro Kinoshita, Shoji Okinaga, Terufumi Kato, Toshiyuki Harada, Akihiko Gemma, Yasuo Saijo, Yuki Yokomizo, Satoshi Morita, Koichi Hagiwara, Toshihiro Nukiwa
    ONCOLOGIST 17 (6) 863 - 870 1083-7159 2012/06 [Refereed][Not invited]
    Background. For non-small cell lung cancer (NSCLC) patients with epidermal growth factor receptor (EGFR) mutations, first-line gefitinib produced a longer progression-free survival interval than first-line carboplatin plus paclitaxel but did not show any survival advantage in the North East Japan 002 study. This report describes the quality of life (QoL) analysis of that study. Methods. Chemotherapy-naive patients with sensitive EGFR-mutated, advanced NSCLC were randomized to receive gefitinib or chemotherapy (carboplatin and paclitaxel). Patient QoL was assessed weekly using the Care Notebook, and the primary endpoint of the QoL analysis was time to deterioration from baseline on each of the physical, mental, and life well-being QoL scales. Kaplan-Meier probability curves and log-rank tests were employed to clarify differences. Results. QoL data from 148 patients (72 in the gefitinib arm and 76 in the carboplatin plus paclitaxel arm) were analyzed. Time to defined deterioration in physical and life well-being significantly favored gefitinib over chemotherapy (hazard ratio [HR] of time to deterioration, 0.34; 95% confidence interval [CI], 0.23-0.50; p < .0001 and HR, 0.43; 95% CI, 0.28-0.65; p < .0001, respectively). Conclusion. QoL was maintained much longer in patients treated with gefitinib than in patients treated with standard chemotherapy, indicating that gefitinib should be considered as the standard first-line therapy for advanced EGFR-mutated NSCLC in spite of no survival advantage. The Oncologist 2012;17:863- 870
  • Sodai Narumi, Akira Inoue, Naoto Morikawa, Yuji Minegishi, Makoto Maemondo, Shoji Okinaga, Kunihiko Kobayashi, Toshiyuki Harada, Koichi Hagiwara, Toshihiro Nukiwa, Akihiko Gemma
    JOURNAL OF CLINICAL ONCOLOGY 30 (15) 0732-183X 2012/05 [Refereed][Not invited]
  • Huqun, Rinako Ishikawa, Jialing Zhang, Hitoshi Miyazawa, Yoshiya Goto, Yoshihiko Shimizu, Koichi Hagiwara, Nobuyuki Koyama
    CANCER 118 (6) 1599 - 1606 0008-543X 2012/03 [Refereed][Not invited]
    BACKGROUND: Enhancer of zeste homolog 2 (EZH2) epigenetically silences many genes through the trimethylation of histone H3 lysine 27 and is implicated in tumor growth, invasion, and metastasis. However, its role in lung cancer has not been well characterized. The objective of the current study was to elucidate the role of EZH2 in nonsmall cell lung cancer (NSCLC) by investigating both clinical samples and cell lines. METHODS: An immunohistochemical analysis of EZH2 expression was performed in samples from patients with stage I NSCLC to investigate the association of EZH2 expression levels with clinicopathologic variables. An in vitro cell growth assay and a Matrigel invasion assay also were conducted in the EZH2-expressing NSCLC cell lines A549 and H1299 after knocking down EZH2 expression by using an EZH2-specific short-hairpin RNA. RESULTS: The immunohistochemical analysis classified stage I NSCLC samples (n 106) into a negative EZH2 expression group (n 40; 37.7%) and a positive EZH2 expression group (n 66; 62.3%). Positive EZH2 expression was associated significantly with larger tumor size (P.014). Kaplan-Meier survival analyses and log-rank tests demonstrated that patients whose samples were classified into the positive EZH2 expression group had a significantly shorter overall survival (P.015). Experiments in the NSCLC cell lines revealed that the knockdown of EZH2 expression reduced the tumor growth rate and invasive activity. CONCLUSIONS: The current results indicated that EZH2 promotes progression and invasion of NSCLC, and its expression is a novel prognostic biomarker in NSCLC. Cancer 2012; 118: 1599-606. VC 2011 American Cancer Society.
  • Takeo Ishii, Koichi Hagiwara, Koichiro Kamio, Shinobu Ikeda, Tomio Arai, Makiko Naka Mieno, Toshio Kumasaka, Masaaki Muramatsu, Motoji Sawabe, Akihiko Gemma, Kozui Kida
    EUROPEAN JOURNAL OF HUMAN GENETICS 20 (2) 230 - 235 1018-4813 2012/02 [Refereed][Not invited]
    Surfactant protein D (SFTPD) induces emphysema in knockout mice, but the association of SFTPD with chronic obstructive pulmonary disease (COPD) and emphysema in humans is unclear. Therefore, we aimed to determine the association between genetic variations in SFTPD and susceptibility to COPD and emphysema. Two populations were studied: population A comprised 270 smokers, including 188 COPD and 82 at-risk subjects, and population B comprised 1131 autopsy cases including 160 cases with emphysema. Six single-nucleotide polymorphisms (SNPs) that tagged the linkage disequilibrium blocks on the entire SFTPD gene were genotyped; the associations of the genotypes with COPD, pulmonary function, percentage of the low-attenuation area (LAA%), and percentage of the airway wall area (WA%) were determined in population A. In population B, the associations of the genotypes with emphysema were assessed. A C allele at SNP rs721917 that results in the replacement of Met with Thr at position 11 in SFTPD was positively correlated with the LAA% in the upper lung (P=1.1x10(-5)) and overall LAA% (P=1.0x10(-4)), and negatively correlated with the serum concentration of SFTPD (P=7x10(-11)) in the population A. The C/C (rs721917/rs10887199) haplotype was associated with emphysema in both the populations. Subjects with a C allele at rs721917 have a lower serum SFTPD concentration and are more susceptible to emphysema. This suggests a protective effect of SFTPD against COPD and emphysema. European Journal of Human Genetics (2012) 20, 230-235; doi:10.1038/ejhg.2011.183; published online 21 September 2011
  • Noriyuki Yamada, Satoshi Oizumi, Hajime Asahina, Naofumi Shinagawa, Eiki Kikuchi, Junko Kikuchi, Jun Sakakibara-Konishi, Tomoaki Tanaka, Kunihiko Kobayashi, Koichi Hagiwara, Masaharu Nishimura
    ONCOLOGY 82 (6) 341 - 346 0030-2414 2012 [Refereed][Not invited]
    Objectives: Cytological examination of samples obtained by bronchoscopy is a useful method for establishing the diagnosis of non-small cell lung cancer (NSCLC). However, the utility of a highly sensitive method for the detection of epidermal growth factor receptor (EGFR) mutation in the cytological specimens has not been fully evaluated. Methods: We retrospectively examined the efficacy of the peptide nucleic acid-locked nucleic acid polymerase chain reaction (PNA-LNA PCR) clamp method for detecting EGFR mutations in 122 bronchoscopic cytological specimens from NSCLC patients. Results: Overall, 41 specimens (33.6%) were positive for EGFR mutation. Twenty-nine (39.7%) of 73 specimens obtained by using endobronchial ultrasonography with a guide sheath, 7 (33.3%) of 21 specimens obtained under direct vision by using a conventional bronchoscope, 4 (36.4%) of 11 specimens obtained by using an ultrathin bronchoscope, and 1 (5.9%) of 17 specimens obtained by endobronchial ultrasound-guided transbronchial needle aspiration were positive for EGFR mutation. Furthermore, among 22 resected NSCLC cases, the EGFR mutation status obtained from bronchoscopic materials was consistent with the status obtained from surgical samples, with the exception of 1 case. Conclusion: The detection of EGFR mutation by subjecting bronchoscopic cytological specimens to a PNA-LNA PCR clamp assay proves useful. Copyright (C) 2012 S. Karger AG, Basel
  • John M. Brehm, Koichi Hagiwara, Yohannes Tesfaigzi, Shannon Bruse, Thomas J. Mariani, Soumyaroop Bhattacharya, Nadia Boutaoui, John P. Ziniti, Manuel E. Soto-Quiros, Lydiana Avila, Michael H. Cho, Blanca Himes, Augusto A. Litonjua, Francine Jacobson, Per Bakke, Amund Gulsvik, Wayne H. Anderson, David A. Lomas, Erick Forno, Soma Datta, Edwin K. Silverman, Juan C. Celedon
    THORAX 66 (12) 1085 - 1090 0040-6376 2011/12 [Refereed][Not invited]
    Rationale Traditional genome-wide association studies (GWASs) of large cohorts of subjects with chronic obstructive pulmonary disease (COPD) have successfully identified novel candidate genes, but several other plausible loci do not meet strict criteria for genome-wide significance after correction for multiple testing. Objectives The authors hypothesise that by applying unbiased weights derived from unique populations we can identify additional COPD susceptibility loci. Methods The authors performed a homozygosity haplotype analysis on a group of subjects with and without COPD to identify regions of conserved homozygosity haplotype (RCHHs). Weights were constructed based on the frequency of these RCHHs in case versus controls, and used to adjust the p values from a large collaborative GWAS of COPD. Results The authors identified 2318 RCHHs, of which 576 were significantly (p<0.05) over-represented in cases. After applying the weights constructed from these regions to a collaborative GWAS of COPD, the authors identified two single nucleotide polymorphisms (SNPs) in a novel gene (fibroblast growth factor-7 (FGF7)) that gained genome-wide significance by the false discovery rate method. In a follow-up analysis, both SNPs (rs12591300 and rs4480740) were significantly associated with COPD in an independent population (combined p values of 7.9E-7 and 2.8E-6, respectively). In another independent population, increased lung tissue FGF7 expression was associated with worse measures of lung function. Conclusion Weights constructed from a homozygosity haplotype analysis of an isolated population successfully identify novel genetic associations from a GWAS on a separate population. This method can be used to identify promising candidate genes that fail to meet strict correction for multiple testing.
  • Yotaro Takaku, Kazuyuki Nakagome, Takehito Kobayashi, Koichi Hagiwara, Minoru Kanazawa, Makoto Nagata
    Respiratory Research 12 138  1465-9921 2011/10 [Refereed][Not invited]
    Background: Eosinophils play an important role in the pathogenesis of bronchial asthma and its exacerbation. Recent reports suggest the involvement of IFN-γ-inducible protein of 10 kDa (IP-10) in virus-induced asthma exacerbation. The objective of this study was to examine whether CXCR3 ligands including IP-10 modify the effector functions of eosinophils.Methods: Eosinophils isolated from the blood of healthy donors were stimulated with CXCR3 ligands and their adhesion to rh-ICAM-1 was then measured using eosinophil peroxidase assays. The generation of eosinophil superoxide anion (O2 -) was examined based on the superoxide dismutase-inhibitable reduction of cytochrome C. Eosinophil-derived neurotoxin (EDN) release was evaluated to determine whether CXCR3 ligands induced eosinophil degranulation. Cytokine and chemokine production by eosinophils was examined using a Bio-plex assay.Results: Eosinophil adhesion to ICAM-1 was significantly enhanced by IP-10, which also significantly induced eosinophil O2 - generation in the presence of ICAM-1. Both the enhanced adhesion and O2 - generation were inhibited by an anti-β2 integrin mAb or an anti-CXCR3 mAb. Other CXCR3 ligands, such as monokine induced by IFN-γ (Mig) and IFN-inducible T cell α chemoattractant (I-TAC), also induced eosinophil adhesion and O2 - generation in the presence of ICAM-1. IP-10, but not Mig or I-TAC, increased the release of EDN. IP-10 increased the production of a number of cytokines and chemokines by eosinophils.Conclusions: These findings suggest that CXCR3 ligands such as IP-10 can directly upregulate the effector functions of eosinophils. These effects might be involved in the activation and infiltration of eosinophils in the airway of asthma, especially in virus-induced asthma exacerbation. © 2011 Takaku et al licensee BioMed Central Ltd.
  • Koichi Hagiwara, Hiroyuki Morino, Jun Shiihara, Tomoaki Tanaka, Hitoshi Miyazawa, Tomoko Suzuki, Masakazu Kohda, Yasushi Okazaki, Kuniaki Seyama, Hideshi Kawakami
    PLOS ONE 6 (9) 1932-6203 2011/09 [Refereed][Not invited]
    Genes involved in disease that are not common are often difficult to identify; a method that pinpoints them from a small number of unrelated patients will be of great help. In order to establish such a method that detects recessive genes identical-by-descent, we modified homozygosity mapping (HM) so that it is constructed on the basis of homozygosity haplotype (HM on HH) analysis. An analysis using 6 unrelated patients with Siiyama-type alpha 1-antitrypsin deficiency, a disease caused by a founder gene, the correct gene locus was pinpointed from data of any 2 patients (length: 1.2-21.8 centimorgans, median: 1.6 centimorgans). For a test population in which these 6 patients and 54 healthy subjects were scrambled, the approach accurately identified these 6 patients and pinpointed the locus to a 1.4-centimorgan fragment. Analyses using synthetic data revealed that the analysis works well for IBD fragment derived from a most recent common ancestor (MRCA) who existed less than 60 generations ago. The analysis is unsuitable for the genes with a frequency in general population more than 0.1. Thus, HM on HH analysis is a powerful technique, applicable to a small number of patients not known to be related, and will accelerate the identification of disease-causing genes for recessive conditions.
  • Takashi Hirama, Takefumi Yamaguchi, Hitoshi Miyazawa, Tomoaki Tanaka, Giichi Hashikita, Etsuko Kishi, Yoshimi Tachi, Shun Takahashi, Keiji Kodama, Hiroshi Egashira, Akemi Yokote, Kunihiko Kobayashi, Makoto Nagata, Toshiaki Ishii, Manabu Nemoto, Masahiko Tanaka, Koichi Fukunaga, Satoshi Morita, Minoru Kanazawa, Koichi Hagiwara
    PLOS ONE 6 (9) e24474  1932-6203 2011/09 [Refereed][Not invited]
    Commensal organisms are frequent causes of pneumonia. However, the detection of these organisms in the airway does not mean that they are the causative pathogens; they may exist merely as colonizers. In up to 50% cases of pneumonia, the causative pathogens remain unidentified, thereby hampering targeting therapies. In speculating on the role of a commensal organism in pneumonia, we devised the battlefield hypothesis. In the "pneumonia battlefield,'' the organism-to-human cell number ratio may be an index for the pathogenic role of the organism. Using real-time PCR reactions for sputum samples, we tested whether the hypothesis predicts the results of bacteriological clinical tests for 4 representative commensal organisms: Streptococcus pneumoniae, Haemophilus influenzae, Pseudomonas spp., and Moraxella catarrhalis. The cutoff value for the organism-to-human cell number ratio, above which the pathogenic role of the organism was suspected, was set up for each organism using 224 sputum samples. The validity of the cutoff value was then tested in a prospective study that included 153 samples; the samples were classified into 3 groups, and each group contained 93%, 7%, and 0% of the samples from pneumonia, in which the pathogenic role of Streptococcus pneumoniae was suggested by the clinical tests. The results for Haemophilus influenzae, Pseudomonas spp., and Moraxella catarrhalis were 100%, 0%, and 0%, respectively. The battlefield hypothesis enabled legitimate interpretation of the PCR results and predicted pneumonia in which the pathogenic role of the organism was suggested by the clinical test. The PCR reactions based on the battlefield hypothesis may help to promote targeted therapies for pneumonia. The prospective observatory study described in the current report had been registered to the University Hospital Medical Information Network (UMIN) registry before its initiation, where the UMIN is a registry approved by the International Committee of Medical Journal Editors (ICMJE). The UMIN registry number was UMIN000001118: A prospective study for the investigation of the validity of cutoff values established for the HIRA-TAN system (April 9, 2008).
  • Hagiwara K
    Nihon Naika Gakkai zasshi. The Journal of the Japanese Society of Internal Medicine 100 (9) 2695 - 2701 0021-5384 2011/09 [Refereed][Not invited]
  • Hironori Satoh, Akira Inoue, Kunihiko Kobayashi, Makoto Maemondo, Satoshi Oizumi, Hiroshi Isobe, Akihiko Gemma, Yasuo Saijo, Hirohisa Yoshizawa, Koichi Hagiwara, Toshihiro Nukiwa
    JOURNAL OF THORACIC ONCOLOGY 6 (8) 1413 - 1417 1556-0864 2011/08 [Refereed][Not invited]
    Introduction: Although standard schedule of gefitinib was the administration of 250 mg tablet every day, many patients need dose reduction because of toxicities. However, the efficacy of such low-dose gefitinib for patients with epidermal growth factor receptor-mutated non-small cell lung cancer has rarely been evaluated. Methods: A post hoc comparison of the efficacy (response rate and survival) in patients treated with gefitinib with or without any dose reduction in NEJ002 study was performed. Results: Among 114 patients treated with first-line gefitinib in NEJ002, 61 (54%) continued gefitinib without any dose reduction until their diseases progressed, and 53 (46%) reduced their dose of gefitinib because of some toxicities. There was no significant difference of patient characteristics between the two groups. The progression-free survival of low-dose group tended to be better than that of standard-dose group (median progression-free survival, 11.8 versus 9.9 months; p = 0.144), and the overall survival of low-dose group was also better than that of standard-dose group (median survival time, 32.7 versus 25.3 months; p = 0.049). Conclusions: The results suggest that low-dose gefitinib may be clinically not inferior to standard-dose gefitinib for non-small cell lung cancer with sensitive epidermal growth factor receptor mutations. Prospective study of low-dose gefitinib is warranted especially for frail patients who need less toxic treatment.
  • Michal Skronski, Tomoaki Tanaka, Adam Szpechcinski, Renata Langfort, Koichi Hagiwara, Tadeusz Orlowski, Joanna Chorostowska-Wynimko
    JOURNAL OF THORACIC ONCOLOGY 6 (6) S999 - S1000 1556-0864 2011/06 [Refereed][Not invited]
  • Kazuyuki Nakagome, Mitsuru Imamura, Hirokazu Okada, Kimito Kawahata, Tsutomu Inoue, Kumiko Hashimoto, Hiroaki Harada, Takehiro Higashi, Rie Takagi, Kazuhisa Nakano, Koichi Hagiwara, Minoru Kanazawa, Makoto Dohi, Makoto Nagata, Sho Matsushita
    JOURNAL OF IMMUNOLOGY 186 (10) 5975 - 5982 0022-1767 2011/05 [Refereed][Not invited]
    Allergic airway inflammation is generally considered a Th2-type immune response. Recent studies, however, demonstrated that Th17-type immune responses also play important roles in this process, especially in the pathogenesis of neutrophilic airway inflammation, a hallmark of severe asthma. We previously reported that dendritic cells release dopamine to naive CD4(+) T cells in Ag-specific cell-cell interaction, in turn inducing Th17 differentiation through dopamine D1-like receptor (D1-like-R). D1-like-R antagonist attenuates Th17-mediated diseases such as experimental autoimmune encephalomyelitis and autoimmune diabetes. However, the effect of antagonizing D1-like-R on Th17-mediated airway inflammation has yet to be studied. In this study, we examined whether D1-like-R antagonist suppresses OVA-induced neutrophilic airway inflammation in OVA TCR-transgenic DO11.10 mice and then elucidated the mechanism of action. DO11.10 mice were nebulized with OVA or PBS, and some mice received D1-like-R antagonist orally before OVA nebulization. D1-like-R antagonist significantly suppressed OVA-induced neutrophilic airway inflammation in DO11.10 mice. It also inhibited the production of IL-17 and infiltration of Th17 cells in the lung. Further, D1-like-R antagonist suppressed the production of IL-23 by lung CD11c(+) APCs. In contrast, D1-like-R antagonist did not increase Foxp3(+) regulatory T cells in the lung. D1-like-R antagonist neither suppressed nonspecific LPS-induced neutrophilic airway inflammation nor OVA-induced eosinophilic airway inflammation. These results indicate that D1-like-R antagonist could suppress Th17-mediated neutrophilic airway inflammation, raising the possibility that antagonizing D1-like-R serves as a promising new strategy for treating neutrophil-dominant severe asthma. The Journal of Immunology, 2011, 186: 5975-5982.
  • Koyama Nobuyuki, Rinako Sadakata, Hitoshi Miyazawa, Koichi Hagiwara, Kunihiko Kobayashi
    CANCER RESEARCH 71 0008-5472 2011/04 [Refereed][Not invited]
  • Ishimoto Osamu, Kunihiko Kobayashi, Akira Inoue, Makoto Maemondo, Shunichi Sugawara, Satoshi Oizumi, Yasuo Saijo, Akihiko Gemma, Satoshi Morita, Koichi Hagiwara, Toshihiro Nukiwa
    ANNALS OF ONCOLOGY 21 11 - 11 0923-7534 2010/11 [Refereed][Not invited]
  • Huqun, Shun-ichiro Fukuyama, Hiroyuki Morino, Hiroshi Miyazawa, Tomoaki Tanaka, Tomoko Suzuki, Masakazu Kohda, Hideshi Kawakami, Yasushi Okazaki, Kuniaki Seyama, Koichi Hagiwara
    BMC BIOINFORMATICS 11 S5  1471-2105 2010/10 [Refereed][Not invited]
    Homozygosity mapping is a powerful procedure that is capable of detecting recessive disease-causing genes in a few patients from families with a history of inbreeding. We report here a homozygosity mapping algorithm for high-density single nucleotide polymorphism arrays that is able to (i) correct genotyping errors, (ii) search for autozygous segments genome-wide through regions with runs of homozygous SNPs, (iii) check the validity of the inbreeding history, and (iv) calculate the probability of the disease-causing gene being located in the regions identified. The genotyping error correction restored an average of 94.2% of the total length of all regions with run of homozygous SNPs, and 99.9% of the total length of them that were longer than 2 cM. At the end of the analysis, we would know the probability that regions identified contain a disease-causing gene, and we would be able to determine how much effort should be devoted to scrutinizing the regions. We confirmed the power of this algorithm using 6 patients with Siiyama-type alpha 1-antitrypsin deficiency, a rare autosomal recessive disease in Japan. Our procedure will accelerate the identification of disease-causing genes using high-density SNP array data.
  • Makoto Maemondo, Akira Inoue, Kunihiko Kobayashi, Shunichi Sugawara, Satoshi Oizumi, Hiroshi Isobe, Akihiko Gemma, Masao Harada, Hirohisa Yoshizawa, Ichiro Kinoshita, Yuka Fujita, Shoji Okinaga, Haruto Hirano, Kozo Yoshimori, Toshiyuki Harada, Takashi Ogura, Masahiro Ando, Hitoshi Miyazawa, Tomoaki Tanaka, Yasuo Saijo, Koichi Hagiwara, Satoshi Morita, Toshihiro Nukiwa
    NEW ENGLAND JOURNAL OF MEDICINE 362 (25) 2380 - 2388 0028-4793 2010/06 [Refereed][Not invited]
    Background: Non-small-cell lung cancer with sensitive mutations of the epidermal growth factor receptor (EGFR) is highly responsive to EGFR tyrosine kinase inhibitors such as gefitinib, but little is known about how its efficacy and safety profile compares with that of standard chemotherapy. Methods: We randomly assigned 230 patients with metastatic, non-small-cell lung cancer and EGFR mutations who had not previously received chemotherapy to receive gefitinib or carboplatin-paclitaxel. The primary end point was progression-free survival; secondary end points included overall survival, response rate, and toxic effects. Results: In the planned interim analysis of data for the first 200 patients, progression-free survival was significantly longer in the gefitinib group than in the standard-chemotherapy group (hazard ratio for death or disease progression with gefitinib, 0.36; P<0.001), resulting in early termination of the study. The gefitinib group had a significantly longer median progression-free survival (10.8 months, vs. 5.4 months in the chemotherapy group; hazard ratio, 0.30; 95% confidence interval, 0.22 to 0.41; P<0.001), as well as a higher response rate (73.7% vs. 30.7%, P<0.001). The median overall survival was 30.5 months in the gefitinib group and 23.6 months in the chemotherapy group (P=0.31). The most common adverse events in the gefitinib group were rash (71.1%) and elevated aminotransferase levels (55.3%), and in the chemotherapy group, neutropenia (77.0%), anemia (64.6%), appetite loss (56.6%), and sensory neuropathy (54.9%). One patient receiving gefitinib died from interstitial lung disease. Conclusions: First-line gefitinib for patients with advanced non-small-cell lung cancer who were selected on the basis of EGFR mutations improved progression-free survival, with acceptable toxicity, as compared with standard chemotherapy. (UMIN-CTR number, C000000376.) N Engl J Med 2010;362:2380-8.
  • Hirofumi Maruyama, Hiroyuki Morino, Hidefumi Ito, Yuishin Izumi, Hidemasa Kato, Yasuhito Watanabe, Yoshimi Kinoshita, Masaki Kamada, Hiroyuki Nodera, Hidenori Suzuki, Osamu Komure, Shinya Matsuura, Keitaro Kobatake, Nobutoshi Morimoto, Koji Abe, Naoki Suzuki, Masashi Aoki, Akihiro Kawata, Takeshi Hirai, Takeo Kato, Kazumasa Ogasawara, Asao Hirano, Toru Takumi, Hirofumi Kusaka, Koichi Hagiwara, Ryuji Kaji, Hideshi Kawakami
    Nature 465 (7295) 223 - 6 0028-0836 2010/05 [Refereed][Not invited]
    Amyotrophic lateral sclerosis (ALS) has its onset in middle age and is a progressive disorder characterized by degeneration of motor neurons of the primary motor cortex, brainstem and spinal cord. Most cases of ALS are sporadic, but about 10% are familial. Genes known to cause classic familial ALS (FALS) are superoxide dismutase 1 (SOD1), ANG encoding angiogenin, TARDP encoding transactive response (TAR) DNA-binding protein TDP-43 (ref. 4) and fused in sarcoma/translated in liposarcoma (FUS, also known as TLS). However, these genetic defects occur in only about 20-30% of cases of FALS, and most genes causing FALS are unknown. Here we show that there are mutations in the gene encoding optineurin (OPTN), earlier reported to be a causative gene of primary open-angle glaucoma (POAG), in patients with ALS. We found three types of mutation of OPTN: a homozygous deletion of exon 5, a homozygous Q398X nonsense mutation and a heterozygous E478G missense mutation within its ubiquitin-binding domain. Analysis of cell transfection showed that the nonsense and missense mutations of OPTN abolished the inhibition of activation of nuclear factor kappa B (NF-kappaB), and the E478G mutation revealed a cytoplasmic distribution different from that of the wild type or a POAG mutation. A case with the E478G mutation showed OPTN-immunoreactive cytoplasmic inclusions. Furthermore, TDP-43- or SOD1-positive inclusions of sporadic and SOD1 cases of ALS were also noticeably immunolabelled by anti-OPTN antibodies. Our findings strongly suggest that OPTN is involved in the pathogenesis of ALS. They also indicate that NF-kappaB inhibitors could be used to treat ALS and that transgenic mice bearing various mutations of OPTN will be relevant in developing new drugs for this disorder.
  • Nobuyuki Koyama, Jialing Zhang, Hu Qun, Yoshiya Goto, Yuichi Ishikawa, Yoshihiko Shimizu, Kozo Sakaguchi, Kunihiko Kobayashi, Koichi Hagiwara
    CANCER RESEARCH 70 0008-5472 2010/04 [Refereed][Not invited]
  • Tomoaki Tanaka, Masaru Matsuoka, Akihisa Sutani, Akihiko Gemma, Makoto Maemondo, Akira Inoue, Shoji Okinaga, Makoto Nagashima, Satoshi Oizumi, Kazutsugu Uematsu, Yoshiaki Nagai, Gaku Moriyama, Hitoshi Miyazawa, Kenji Ikebuchi, Satoshi Morita, Kunihiko Kobayashi, Koichi Hagiwara
    INTERNATIONAL JOURNAL OF CANCER 126 (3) 651 - 655 0020-7136 2010/02 [Refereed][Not invited]
    Mutation in the epidermal growth factor receptor (EGFR) is frequently seen in non-small cell lung cancers (NSCLCs), especially in Asian females with adenocarcinoma. The frequency of mutation and the factors associated requires to be elucidated by analyzing a large number of consecutive clinical samples. We summarized the result of the EGFR mutation analysis for 1,176 patients performed at the time of diagnosis or relapse. The PNA-LNA PCR clamp, a highly sensitive detection method for the EGFR mutation, was employed. For fresh cases a portion of samples isolated to establish the diagnosis of lung cancer was used. For cases with a relapsed disease archival tissue were tested. The variables associated with the EGFR mutation after removing the confound factors were investigated by the logistic analysis using the samples collected in our university (n = 308) where detailed information on patients were available. The frequency of the EGFR mutation and its subtypes were investigated using all samples (n = 1,176). The EGFR mutation was significantly associated with adenocarcinoma (P = 0.006) and light-smoking (p < 0.0001), but not gender. The deletions in exon 19 were more frequently associated with mate gender while exon 21 deletions were with female gender (p = 0.0011). The overall frequency of the EGFR mutation was 31%. Our result suggests that the female predominance in the EGFR mutation rate is a reflection of a higher frequency of adenocarcinoma in females. The gender difference in the mutation subtypes may provide a clue for the mechanism of the occurrence of the EGFR mutation.
  • Takefumi Yamaguchi, Tomoyuki Soma, Yotaro Takaku, Kazuyuki Nakagome, Koichi Hagiwara, Minoru Kanazawa, Makoto Nagata
    INTERNATIONAL ARCHIVES OF ALLERGY AND IMMUNOLOGY 152 32 - 40 1018-2438 2010 [Refereed][Not invited]
    Background: There is evidence that excessive use of inhalational beta(2)-agonists induces the deterioration of asthma. Although the exact mechanism of this remains to be elucidated, overuse of beta(2)-agonists may impair the Th1/Th2 balance in asthmatic airways. The aim of the present study was to evaluate whether salbutamol, a representative inhalational beta(2)-agonist, modifies the production of Th1- and Th2-type cytokines by mononuclear cells separated from patients with asthma and healthy volunteers. Methods: Peripheral blood mononuclear cells (PBMCs) obtained from 8 healthy volunteers and 10 patients with mild persistent asthma allergic to house dust mites were treated with either salbutamol or medium alone. PBMCs were then stimulated with either medium alone, house dust mite (Dermatophagoides farina, Df) allergen or a combination of ionomycin plus phorbol 12-myristate 13-acetate ester (PMA). Concentrations of IFN-gamma, IL-13, TNF-alpha and RANTES in the cell supernatants were measured using ELISA. Results: In PBMCs from healthy volunteers, salbutamol did not modify IFN-gamma production, but increased the spontaneous production of IL-13. In contrast, salbutamol significantly inhibited the spontaneous and ionomycin- plus PMA-stimulated production of IFN-gamma by PBMCs from asthmatics. Salbutamol significantly enhanced both spontaneous and Df-induced production of IL-13 by PBMCs from asthmatics. Salbutamol did not modify the production of TNF-alpha. Finally, salbutamol enhanced the production of RANTES induced by Df allergen in asthmatics. Conclusions: Salbutamol inhibits IFN-gamma and enhances IL-13 production by PBMCs from asthmatics. These effects would promote a Th1/Th2 imbalance in the airways and may therefore contribute to the deterioration of asthma. Copyright (C) 2010 S. Karger AG, Basel
  • Yotaro Takaku, Kazuyuki Nakagome, Takehito Kobayashi, Takefumi Yamaguchi, Fuyumi Nishihara, Tomoyuki Soma, Koichi Hagiwara, Minoru Kanazawa, Makoto Nagata
    INTERNATIONAL ARCHIVES OF ALLERGY AND IMMUNOLOGY 152 41 - 46 1018-2438 2010 [Refereed][Not invited]
    Background: Most patients with asthma are currently controlled by pharmacotherapeutic means such as inhaled corticosteroid (ICS). However, whether ICS actually induces remission of asthma remains unknown. The present study evaluates changes in airway inflammation and hyperresponsiveness in adult patients with asthma after stopping ICS. Methods: We enrolled 11 patients with allergic asthma (7 males and 4 females; mean age, 52.3 years) who had been asymptomatic and had no exacerbation by low-dose ICS. Airway hyperresponsiveness (AHR) was assessed using methacholine challenge, and induced sputum was evaluated before and every 3 months after ICS cessation during the 1-year follow-up. Results: Among the 11 asthmatics, AHR increased in 10 (90.9%) and asthma clinically relapsed in 4 (36.4%) within 1 year of ICS cessation. AHR increased in all 7 asthmatics that were sensitized to Dermatophagoides farinae and asthma clinically relapsed in 4 (57.1%) of them. Furthermore, eosinophil numbers and IL-4 concentrations in the sputum significantly increased after ICS cessation. Conclusions: Remission with normal airway response to methacholine (no AHR) might be rare in adult patients with allergic asthma, and sensitization to house dust mites appears to play an important role in relapse. Therefore, ICS cessation should be carefully considered in patients sensitive to house dust mites. Serial determination of eosinophil counts or IL-4 concentrations in sputum might be appropriate for monitoring and preventing asthma relapse in adults. Copyright (C) 2010 S. Karger AG, Basel
  • Rinako Sadakata, Atsushi Hatamochi, Keiji Kodama, Akiko Kaga, Takefumi Yamaguchi, Tomoyuki Soma, Yutaka Usui, Makoto Nagata, Akira Ohtake, Koichi Hagiwara, Minoru Kanazawa
    INTERNAL MEDICINE 49 (16) 1797 - 1800 0918-2918 2010 [Refereed][Not invited]
    Ehlers-Danlos syndrome type IV (EDS type IV), vascular type, an autosomal dominant disorder caused by a mutation of the type III procollagen gene (COL3A1) is the most severe form of EDS and often presents with aortic hemorrhage or organ perforation. This report discusses a male patient with EDS type IV with dyspnea due to hemopneumothorax. He had thin skin and hypermobile joints and was clinically confirmed as having EDS type IV. The diagnosis was genetically confirmed by a mutation c. 2528 G>A (p.Gly843Glu) in the COL3A1 gene. The position of the mutation has never been reported.
  • Teruo Tachibana, Koichi Hagiwara, Takeshi Johkoh
    CURRENT OPINION IN PULMONARY MEDICINE 15 (5) 486 - 490 1070-5287 2009/09 [Refereed][Not invited]
    Purpose of review Our knowledge of pulmonary alveolar microlithiasis (PAM) has significantly increased since its detailed description by Sosman in 1957. Here we provide updated information on the long-term clinical course, the specific findings in imaging studies and the genetics of this disease. Recent findings The responsible gene, the mutation of which is associated with PAM, has been identified as SLC34A2. Characteristic chest computed tomography (CT) findings in patients with PAM have been shown to correlate well with specific pathological findings. Elevated serum levels of surfactant proteins A and D have also been reported in this disease. Long-term follow up information has been updated. Summary The gene responsible for PAM, SLC34A2, has been identified. It encodes a type IIb sodium-dependent phosphate transporter, the function of which provides an insight into the pathogenesis of this disease. The demonstration of a mutation in the SLC34A2 gene helps to confirm the diagnosis of PAM. Characteristic chest CT findings that include irregular thickening of perilobular interstitium and calcification along perilobular structures correlate with specific pathological findings. Serum levels of surfactant proteins A and D correlate with the progression of the disease, and may be a useful monitoring tool. Scrutiny of the long-term follow-up data of PAM patients reveals that the prognosis for PAM is poor. The establishment of an effective treatment, which is not yet available, is mandatory.
  • Taro Yokoyama, Nobuyuki Koyama, Keiji Kodama, Koichi Hagiwara, Minoru Kanazawa
    BMJ Case Reports 2009 1757-790X 2009/07 [Refereed][Not invited]
  • Satoshi Morita, Isamu Okamoto, Kunihiko Kobayashi, Koichi Yamazaki, Hajime Asahina, Akira Inoue, Koichi Hagiwara, Noriaki Sunaga, Noriko Yanagitani, Toyoaki Hida, Kimihide Yoshida, Tomonori Hirashima, Kosei Yasumoto, Kenji Sugio, Tetsuya Mitsudomi, Masahiro Fukuoka, Toshihiro Nukiwa
    CLINICAL CANCER RESEARCH 15 (13) 4493 - 4498 1078-0432 2009/07 [Refereed][Not invited]
    Purpose: Somatic mutations of the epidermal growth factor receptor (EGFR) gene are associated with an increased response to gefitinib in patients with non-small cell lung cancer. We have examined the impact of gefitinib on progression-free survival and overall survival in patients with EGFR mutation - positive non - small cell lung cancer. Experimental Design: We searched for all clinical trials that prospectively evaluated the efficacy of gefitinib for advanced non - small cell lung cancer with EGFR mutations in Japan. We did a combined analysis based on individual patient data from the identified trials. Results: Seven eligible trials were identified fora total of 148 non - small cell lung cancer patients with EGFR mutations. The overall response rate to gefitinib was 76.4% [95% confidence interval (95% CI), 69.5-83.2]. The median progression-free survival and overall survival were 9.7 months (95% CI, 8.2-11.1) and 24.3 months (95% CI, 19.8-28.2), respectively. Good performance status and chemotherapy-naive status were significantly associated with a longer progression-free survival or overall survival. Of the 148 patients, 87 received gefitinib as a first-line therapy, whereas 61 received systemic chemotherapy before gefitinib treatment. The median progression-free survival after the start of first-line therapy was significantly longer in the gefitinib-first group than in the chemotherapy-first group (10.7 versus 6.0 months; P < 0.001), whereas no significant difference in median overall survival was apparent between the two groups (27.7 versus 25.7 months; P = 0.782). Conclusions: Gefitinib monotherapy confers substantial clinical benefit in terms of progression-free survival and overall survival in non-small cell lung cancer patients with EGFR mutations. Randomized trials comparing chemotherapy with gefitinib as a first-line treatment are warranted in such patients.
  • Nobuyuki Koyama, Hu Qun, Zialing Zhang, Yoshiya Goto, Hiroyuki Nitanda, Yoshihiko Shimizu, Koichi Kaneko, Minoru Kanazawa, Kunihiko Kobayashi, Koichi Hagiwara
    CANCER RESEARCH 69 0008-5472 2009/05 [Refereed][Not invited]
  • Akira Inoue, Kunihiko Kobayashi, Kazuhiro Usui, Makoto Maemondo, Shoji Okinaga, Iwao Mikami, Masahiro Ando, Koichi Yamazaki, Yasuo Saijo, Akihiko Gemma, Hitoshi Miyazawa, Tomoaki Tanaka, Kenji Ikebuchi, Toshihiro Nukiwa, Satoshi Morita, Koichi Hagiwara
    JOURNAL OF CLINICAL ONCOLOGY 27 (9) 1394 - 1400 0732-183X 2009/03 [Refereed][Not invited]
    Purpose This multicenter phase II study was undertaken to investigate the efficacy and feasibility of gefitinib for patients with advanced non-small-cell lung cancer (NSCLC) harboring epidermal growth factor receptor (EGFR) mutations without indication for chemotherapy as a result of poor performance status (PS). Patients and Methods Chemotherapy-naive patients with poor PS (patients 20 to 74 years of age with Eastern Cooperative Oncology Group PS 3 to 4, 75 to 79 years of age with PS 2 to 4, and >= 80 years of age with PS 1 to 4) who had EGFR mutations examined by the peptide nucleic acid-locked nucleic acid polymerase chain reaction clamp method were enrolled and received gefitinib (250 mg/d) alone. Results Between February 2006 and May 2007, 30 patients with NSCLC and poor PS, including 22 patients with PS 3 to 4, were enrolled. The overall response rate was 66% (90% CI, 51% to 80%), and the disease control rate was 90%. PS improvement rate was 79% (P < .00005); in particular, 68% of the 22 patients improved from >= PS 3 at baseline to <= PS 1. The median progression-free survival, median survival time, and 1-year survival rate were 6.5 months, 17.8 months, and 63%, respectively. No treatment-related deaths were observed. Conclusion This is the first report indicating that EGFR mutation-positive patients with extremely poor PS benefit from first-line gefitinib. Because there previously has been no standard treatment for these patients with short life expectancy other than best supportive care, examination of EGFR mutations as a biomarker is recommended in this patient population.
  • Shinya Kikuchi, Izumi Kikuchi, Yotaro Takaku, Takehito Kobayashi, Koichi Hagiwara, Minoru Kanazawa, Makoto Nagata
    INTERNATIONAL ARCHIVES OF ALLERGY AND IMMUNOLOGY 149 87 - 93 1018-2438 2009 [Refereed][Not invited]
    Background: There is evidence that eosinophils and neutrophils are simultaneously increased in the airways of some patients with chronic refractory asthma. The mechanisms by which neutrophils accumulate in the airways of asthmatics remain to be elucidated, however, chemoattractants for neutrophils such as CXC chemokines may affect either the accumulation or functional status of neutrophils in such patients. The objective of the present study was to identify the CXC chemokine responsible for the neutrophilic and possibly eosinophilic inflammation observed in the airways of patients with refractory asthma. Methods: Following the inhalation of hypertonic saline, induced sputum was obtained from 14 healthy controls, 16 patients with mild well-controlled nonrefractory asthma, and 14 patients with refractory asthma. Concentrations of CXC chemokines and differential inflammatory cell counts were determined. Results: The percentages of induced sputum eosinophils were significantly higher both in patients with nonrefractory asthma and in patients with refractory asthma. On the other hand, the percentages of neutrophils were increased only in sputum from patients with refractory asthma. The concentration of IL-8, but not ENA-78 or GRO-alpha, was also significantly increased in induced sputum from patients with refractory asthma. The concentration of IL-8 correlated significantly with the percentages of neutrophils. Conclusions: The results of the present study suggest that IL-8, but not ENA-78 or GRO-alpha, may contribute to the observation of neutrophilic inflammation in patients with refractory asthma. Copyright (c) 2009 S. Karger AG, Basel
  • Mariko Mori, Yotaro Takaku, Takehito Kobayashi, Koichi Hagiwara, Minoru Kanazawa, Makoto Nagata
    INTERNATIONAL ARCHIVES OF ALLERGY AND IMMUNOLOGY 149 31 - 38 1018-2438 2009 [Refereed][Not invited]
    Rationale: Eosinophils preferentially accumulate at sites of inflammation in the asthmatic airway. Participation of circulating eosinophils in the airway inflammation in asthma involves their interaction with adhesion molecules expressed on the endothelial cell surface and exposure to inflammatory mediators, such as cysteinyl leukotrienes (cysLTs). Objective: To investigate whether interaction of eosinophils with adhesion molecules modifies the functions of these cells induced by cysLTs. Methods: Eosinophils were isolated from the blood of healthy donors, incubated in the EIA plates coated with adhesion proteins, and then exposed to LTD4. The generation of superoxide anion (O-2(-)), adhesion to the plates, and release of eosinophil-derived neutrotoxin (EDN) were evaluated. Results: Neither VCAM-1 nor LTD 4 (100 nM) independently induced eosinophil O-2(-) generation, however, combined exposure to the two molecules synergistically induced eosinophil O-2(-) generation. ICAM-1 by itself induced eosinophil O-2(-) generation, which was enhanced by LTD4. On the contrary, P-selectin did not induce O-2(-) generation, either in the presence or absence of LTD4. LTD4 significantly enhanced eosinophil adhesion to rh-VCAM-1 and rh-ICAM-1, but not to rh-P-selectin. Finally, we observed that combined exposure of eosinophils to LTD4 and VCAM-1 induced the release of EDN. Conclusion: Combined exposure to VCAM-1 or ICAM-1 and cysLT effectively induces the effector functions of eosinophils. Eosinophil adhesion to and migration across endothelial cells via these specific adhesion proteins and subsequent exposure to cysLTs may be mechanisms underlying activation of the effector functions of eosinophils in the asthmatic airway. Copyright (c) 2009 S. Karger AG, Basel
  • Arata Azuma, Koichi Hagiwara, Shoji Kudoh
    AMERICAN JOURNAL OF RESPIRATORY AND CRITICAL CARE MEDICINE 177 (12) 1397 - 1398 1073-449X 2008/06 [Refereed][Not invited]
  • Hitoshi Miyazawa, Tomoaki Tanaka, Yoshiaki Nagai, Masaru Matsuoka, Hu qun, Akihisa Sutani, Kiyoshi Udagawa, Jialing Zhang, Takashi Hirama, Yoshitake Murayama, Nobuyuki Koyama, Kenji Ikebuchi, Makoto Nagata, Minoru Kanazawa, Toshihiro Nukiwa, Seiichi Takenoshita, Kunihiko Kobayashi, Koichi Hagiwara
    CANCER SCIENCE 99 (3) 595 - 600 1347-9032 2008/03 [Refereed][Not invited]
    Mutations in the epidermal growth factor receptor (EGFR) are observed in a fraction of non-small-cell lung cancers (NSCLS). EGFR mutation-positive NSCLS responds to gefitinib. Secondary T790M mutation confers gefitinib resistance to NSCLS. A detection test for the T790M mutation was designed based on the peptide nucleic acid-locked nucleic acid polymerase chain reaction clamp method. The specificity and sensitivity of the test were both greater than 0.99. The test revealed that only a small population of the PC-13 cells carried the T790M mutation. The test also revealed that the T790M mutation was found in none of 151 NSCLC specimens obtained before gefitinib treatment, whereas it was found in four of four specimens obtained from NSCLS that had become refractory to gefitinib. In one patient in whom the L858R-positive EGFR allele was amplified to multiple copies, an L858R-T790M double-mutant allele emerged during the gefitinib therapy. This allele was expressed highly. The T790M mutation detection test based on the peptide nucleic acid-locked nucleic acid polymerase chain reaction clamp method is sensitive and specific, and is applicable to clinical practice. It detects T790M-positive cells in the course of gefitinib treatment, and thus will help to devise therapies effective for T790M-positive NSCLS.
  • Nobuyuki Koyama, Makoto Nagata, Koichi Hagiwara, Minoru Kanazawa
    RESPIROLOGY 13 (2) 309 - 311 1323-7799 2008/03 [Refereed][Not invited]
    Mucormycosis is an uncommon fungal infection, which generally develops in immunosuppressed hosts. In particular, pulmonary infection by Cunninghamella bertholletiae, a rare species of Mucor, is characterized by invasiveness and high mortality. Herein a case of pulmonary mucormycosis due to C. bertholletiae in a female patient with chronic renal insufficiency, secondary to microscopic polyarteritis, is reported. The patient survived after successful treatment with a cumulative dose of 1508 mg of amphotericin B, phased reduction of glucocorticoid therapy and chest tube drainage of a pneumothorax, without the necessity for surgical intervention. This case demonstrates that conservative therapy may be effective in patients for whom surgical intervention is not an option.
  • Akihiko Miyanaga, Akihiko Gemma, Masahiro Ando, Seiji Kosaihira, Rintaro Noro, Yuji Minegishi, Kiyoko Kataoka, Michiya Nara, Tetsuya Okano, Hitoshi Miyazawa, Tomoaki Tanaka, Akinobu Yoshimura, Kunihiko Kobayashi, Hiroshi Iwanami, Koichi Hagiwara, Eitaka Tsuboi, Shoji Kudoh
    ONCOLOGY REPORTS 19 (2) 377 - 383 1021-335X 2008/02 [Refereed][Not invited]
    It is known that an epidermal growth factor receptor (EGFR) gene mutation(s) is present in a percentage of non-small cell lung cancers (NSCLCs). Gefitinib, an inhibitor of the tyrosine kinase activity of EGFR, is effective on most of them. The EGFR mutation status alone cannot fully predict the response to gefitinib and the prognosis for the patients. We hypothesized that information on the expression levels of phosphorylated-EGFR and -Akt, and E-cadherin, alone or in combination with information on the EGFR mutation, may refine our ability of prediction. We investigated 24 NSCLCs that had recurred after surgery and were treated with gefitinib. Specimens resected by surgery were subjected to the peptide nucleic acid-locked nucleic acid polymerase chain reaction clamp reaction to determine the EGFR mutation status, and to immunohistochemical staining of phosphorylated-EGFR and -Akt, and E-cadherin to determine their expression levels. The EGFR mutation status was predictive of responsive disease (complete response: CR + partial response: PR) and controlled disease (CR + PR + stable disease: SD). Positive E-cadherin staining was predictive of longer time to progression (12.4 vs. 5.9 months, p<0.05) and overall survival (OS) (18.4 vs. 13.0 months, p<0.05). Together the patients with an EGFR mutation and the patients with positive E-cadherin staining defined a patient group with a median OS of 18.4 months and excluded the patient group with the median OS of 3.7 months. Neither p-Akt nor p-EGFR staining was associated with the response and survival. In patients with surgically resected NSCLC tumors, the EGFR mutation status and E-cadherin staining can select patients who will benefit from gefitinib therapy.
  • Tomoyuki Soma, Yotaro Takaku, Takehito Kobayashi, Koichi Hagiwara, Minoru Kanazawa, Kazutsugu Uematsu, Makoto Nagata
    INTERNATIONAL ARCHIVES OF ALLERGY AND IMMUNOLOGY 146 22 - 27 1018-2438 2008 [Refereed][Not invited]
    Background: The use of a budesonide (BUD)/formoterol (FOR) combination (Symbicort(R)) for both maintenance and reliever therapy reduces the exacerbation of asthma better than the traditional fixed-dose regimens. In the early stage of exacerbation, the combination therapy could intervene with the development of subsequent airway inflammation. The objective of the study was to evaluate whether in the early stage of cell activation the use of BUD/FOR combination would modify the adhesion of eosinophils or production of cytokines from mononuclear cells. Methods: Human peripheral blood eosinophils, pretreated with BUD (0.1 mu M), FOR (0.1 mu M) or BUD/FOR combination for 30 min at 37 degrees C, were stimulated with IL-5, leukotriene D4 or phorbol myristate acetate, and eosinophil adhesion was evaluated using an eosinophil peroxidase assay. BUD (0.1 mu M), FOR (0.1 mu M) or BUD/FOR combination was added to the peripheral blood mononuclear cells stimulated with ionomycin plus phorbol myristate acetate. Concentrations of IL-5 and RANTES in the cell supernatant were measured using ELISA. Results: BUD, FOR or BUD/FOR combination did not modify the eosinophil adhesion induced by any stimulators. In contrast, BUD or BUD/FOR combination significantly reduced the productions of IL-5 and RANTES in peripheral blood mononuclear cells (BUD: p < 0.0001, p = 0.027 vs. control; BUD/ FOR: p < 0.0001, p = 0.031 vs. control) when the drugs were added 15 min, but not 4 h, following cell stimulation. Conclusion: In the early stage of exacerbation of asthma, inhaled BUD may suppress the progression of the allergic inflammatory cascade by inhibiting the mononuclear cells. These results also underline the importance of using BUD in rescue inhaler. Copyright (C) 2008 S. Karger AG, Basel.
  • Hagiwara K
    Nihon rinsho. Japanese journal of clinical medicine 65 Suppl 8 405 - 409 0047-1852 2007/10 [Refereed][Not invited]
  • Hitoshi Miyazawa, Masaaki Kato, Takuya Awata, Masakazu Kohda, Hiroyasu Iwasa, Nobuyuki Koyama, Tomoaki Tanaka, Huqun, Shunei Kyo, Yasushi Okazaki, Koichi Hagiwara
    AMERICAN JOURNAL OF HUMAN GENETICS 80 (6) 1090 - 1102 0002-9297 2007/06 [Refereed][Not invited]
    A promising strategy for identifying disease susceptibility genes for both single- and multiple-gene diseases is to search patients' autosomes for shared chromosomal segments derived from a common ancestor. Such segments are characterized by the distinct identity of their haplotype. The methods and algorithms currently available have only a limited capability for determining a high-resolution haplotype genomewide. We herein introduce the homozygosity haplotype (HH), a haplotype described by the homozygous SNPs that are easily obtained from high-density SNP genotyping data. The HH represents haplotypes of both copies of homologous autosomes, allowing for direct comparisons of the autosomes among multiple patients and enabling the identification of the shared segments. The HH successfully detected the shared segments from members of a large family with Marfan syndrome, which is an autosomal dominant, single- gene disease. It also detected the shared segments from patients with model multigene diseases originating with common ancestors who lived 10-25 generations ago. The HH is therefore considered to be useful for the identification of disease susceptibility genes in both single- and multiple-gene diseases.
  • Sato N, Sutani A, Oya H, Yamaguchi T, Saito K, Kobayashi K, Nagata M, Hagiwara K, Kanazawa M
    Nihon Kokyuki Gakkai zasshi = the journal of the Japanese Respiratory Society 45 (3) 237 - 242 1343-3490 2007/03 [Refereed][Not invited]
  • Huqun, Shinyu Izumi, Hitoshi Miyazawa, Kuniaki Ishii, Bine Uchiyama, Tadashi Ishida, Sawako Tanaka, Ryushi Tazawa, Shunichiro Fukuyama, Tomoaki Tanaka, Yoshiaki Nagai, Akemi Yokote, Hiroki Takahashi, Toshihiko Fukushima, Kunihiko Kobayashi, Hirofumi Chiba, Makoto Nagata, Susumu Sakamoto, Koichiro Nakata, Yuji Takebayashi, Yoshihiko Shimizu, Koichi Kaneko, Michio Shimizu, Minoru Kanazawa, Shosaku Abe, Yoshikazu Inoue, Seiichi Takenoshita, Kunihiko Yoshimura, Koichiro Kudo, Teruo Tachibana, Toshihiro Nukiwa, Koichi Hagiwara
    AMERICAN JOURNAL OF RESPIRATORY AND CRITICAL CARE MEDICINE 175 (3) 263 - 268 1073-449X 2007/02 [Refereed][Not invited]
    Rationale: Pulmonary alveolar microlithiasis is an autosomal recessive disorder in which microliths are formed in the alveolar space. Objectives: To identify the responsible gene that causes pulmonary alveolar microlithiasis. Methods: By means of a genomewide single-nucleotide polymorphism analysis using DNA from three patients, we have narrowed the region in which the candidate gene is located. From this region, we have identified a gene that has mutations in all patients with pulmonary alveolar microlithiasis. Measurements and Main Results: We identified a candidate gene, SLC34A2, that encodes a type IIb sodium phosphate cotransporter and that is mutated in six of six patients investigated. SLC34A2 is specifically expressed in type 11 alveolar cells, and the mutations abolished the normal gene function. Conclusion: Mutations in the SLC34A2 gene that abolish normal gene function cause pulmonary alveolar microlithiasis.
  • Tomoaki Tanaka, Yoshiaki Nagai, Hitoshi Miyazawa, Nobuyuki Koyama, Suguru Matsuoka, Akihisa Sutani, Huqun, Kiyoshi Udagawa, Yoshitake Murayama, Makoto Nagata, Yoshihiko Shimizu, Kenji Ikebuchi, Minoru Kanazawa, Kunihiko Kobayashi, Koichi Hagiwara
    CANCER SCIENCE 98 (2) 246 - 252 1347-9032 2007/02 [Refereed][Not invited]
    Gefitinib is an inhibitor of the tyrosine kinase activity of epidermal growth factor receptor (EGFR). Accumulating evidence suggests that gefitinib may provide a survival benefit to EGFR mutation-positive non-small lung cancer patients. We have established a clinical test that can detect EGFR mutations from cytological specimens or paraffin-embedded tissue specimens that are contaminated by normal cells. This test is based on the peptide nucleic acid, locked nucleic acid polymerase chain reaction clamp method that can detect G719S, G719C, L858R, L861Q and seven different exon 19 deletions in the presence of 100-1000-fold wild-type alleles. Consequently, using a small aliquot of samples isolated to establish a cancer diagnosis, the EGFR mutation status is determined soon after the diagnosis of cancer is made. We investigated the EGFR mutation status in 86 patients using a variety of cytological specimens (59 bronchoscopy specimens, 16 pleural effusion, 9 sputum, and 2 pericardial effusion) and in 46 patients who had a disease relapse and paraffin-embedded tissues were available. Forty-five patients (34%) were positive for mutation (29 exon 19 deletions, 16 L858R and 1 L861Q). The sensitivity and the specificity of this test was 97% and 100%, respectively. EGFR mutation status thereby obtained was used to determine each patient's therapeutic regimen. This test is easily integrated into the normal clinical practice for lung cancer, while allowing the medical staff to select therapeutic regimen depending on the EGFR mutation status.
  • Takehito Kobayashi, Yotaro Takaku, Izumi Kikuchi, Tomoyuki Soma, Koichi Hagiwara, Minoru Kanazawa, Makoto Nagata
    INTERNATIONAL ARCHIVES OF ALLERGY AND IMMUNOLOGY 143 38 - 43 1018-2438 2007 [Refereed][Not invited]
    Background: There is increasing evidence that both neutrophilic and eosinophilic inflammation persist in the airways of patients with severe asthma. We have reported a positive relationship between the concentrations of eosinophils and neutrophils in sputum from severe asthmatics, suggesting a possible role of eosinophils in regulating neutrophilic inflammation. The aim of this study was to investigate whether activated eosinophils modify the trans-basement membrane migration (TBM) of neutrophils. Methods: Eosinophils and neutrophils were isolated from peripheral blood drawn from healthy donors. The TBM of neutrophils in response to a variety of chemoattractants was evaluated in the presence or absence of eosinophils by using the chambers with a Matrigel (R)-coated Transwell (R) insert. Results: As expected, eotaxin (10 nM) and RANTES (10 nM), but not IL-8 (10 nM), induced the TBM of eosinophils. On the contrary, only IL-8 induced the TBM of neutrophils. When eosinophils were coincubated with neutrophils and stimulated with IL-8, the TBM of eosinophils was significantly augmented. On the other hand, when neutrophils were coincubated with eosinophils and stimulated with eotaxin or RANTES, the TBM of neutrophils was not modified. Conclusions: Neutrophils migrated by IL-8 may lead eosinophils to accumulate in the airways of patients with severe asthma. On the other hand, it is unlikely that eosinophils migrated by chemoattractants such as CC chemokines regulate neutrophilic inflammation. Copyright (C) 2007 S. Karger AG, Basel.
  • Izumi Kikuchi, Shinya Kikuchi, Takehito Kobayashi, Yotaro Takaku, Koichi Hagiwara, Minoru Kanazawa, Makoto Nagata
    INTERNATIONAL ARCHIVES OF ALLERGY AND IMMUNOLOGY 143 44 - 49 1018-2438 2007 [Refereed][Not invited]
    Background: Recent evidence suggests that both neutrophilic and eosinophilic inflammation persist in the airways of patients with severe asthma. Neutrophils can secrete a variety of mediators which may augment the migration of eosinophils. We have reported that activated neutrophils augment the trans-basement membrane migration (TBM) of eosinophils in vitro. Theophylline has been shown to modulate some functions of both neutrophils and eosinophils. The objective of this study was to evaluate whether theophylline modulates the neutrophil-dependent augmentation of eosinophil TBM. Methods: Eosinophils and neutrophils were isolated from peripheral blood collected from healthy donors and were then preincubated with either 0.1 mM theophylline or the medium control. The TBM of eosinophils in response to IL-8 was evaluated in the presence or absence of neutrophils by using the chambers with a Matrigel (R)-coated Transwell (R) insert. The generation of O-2(-) was evaluated by the cytochrome c reduction assay. Results: As previously reported, IL-8-stimulated neutrophils significantly augmented the TBM of eosinophils. Theophylline significantly attenuated the neutrophil-dependent augmentation of eosinophil TBM (p < 0.001) and did not directly modify the TBM of neutrophils in response to IL-8 or LTB4. Similarly, the LTB4-induced TBM of eosinophils was not modified by theophylline. Finally, theophylline attenuated the superoxide anion generation from IL-8-stimulated neutrophils on the Matrigel-coated plates. Conclusions: Our results show that theophylline can attenuate the neutrophil-dependent augmentation of eosinophil TBM. This effect is possibly attributable to the suppression of neutrophil activation provoked by the combination of basement membrane and IL-8. Copyright (C) 2007 S. Karger AG, Basel.
  • Nobuyuki Koyama, Yasuto Jinn, Kazuhiko Takabe, Masafumi Yoshizawa, Yutaka Usui, Naohiko Inase, Shuji Miyake, Yasuyuki Yoshizawa, Koichi Hagiwara, Minoru Kanazawa
    ANTICANCER RESEARCH 26 (6B) 4519 - 4525 0250-7005 2006/11 [Refereed][Not invited]
    Background: Factors predicting gefitinib sensitivity and adverse events in non-small cell lung cancer (NSCLC) remain controversial. Patients and Methods: Correlations among clinicopathological characteristics, gefitinib sensitivity and adverse events were studied in 154 patients with NSCLC, whereas epidermal growth factor receptor (EGFR) mutations were analyzed in 44 patients. Results: Female, non-smoker, adenocarcinoma of stage I-II, and gefitinib effectiveness correlated with longer time to progression (TTP) and overall survival (OS), while the rate of interstitial lung disease in patients undergoing thoracic radiotherapy and stomatitis in females or those who never smoked were significantly higher. EGFR mutations were identified in 18 cases, and among 34 gefitinib-treated patients, 16 patients harboring mutations tended to do better, both in terms of TTP and OS. The results of the mutation analysis from surgical and non-surgical specimens were identical. Conclusion: Certain clinicopathological characteristics and EGFR mutations can be either predictive of gefitinib sensitivity or adverse events.
  • Hagiwara K
    Nihon Naika Gakkai zasshi. The Journal of the Japanese Society of Internal Medicine 95 (6) 1036 - 1041 0021-5384 2006/06 [Refereed][Not invited]
  • Kikuchi, I, S Kikuchi, T Kobayashi, K Hagiwara, Y Sakamoto, M Kanazawa, M Nagata
    AMERICAN JOURNAL OF RESPIRATORY CELL AND MOLECULAR BIOLOGY 34 (6) 760 - 765 1044-1549 2006/06 [Refereed][Not invited]
    Neutrophilic inflammation observed with severe asthma is often associated with interleukin-8 (IL-8). Neutrophils can secrete a variety of mediators that may augment the migration of eosinophils. We have reported a positive correlation between the concentrations of neutrophils and eosinophils in sputum from subjects with severe asthma, suggesting a possible role of neutrophils in regulating eosinophilic inflammation. The aim of this study was to investigate whether neutrophils stimulated with IL-8 modify the trans-basement membrane migration (TBM) of eosinophils. Eosinophils and neutrophils were isolated from peripheral blood drawn from healthy donors or subjects with mild asthma. The TBM of eosinophils in response to IL-8 was evaluated in the presence or absence of neutrophils using the chambers with a Matrigel-coated transwell insert. Neither IL-8 alone nor the presence of neutrophils alone induced the TBM of eosinophils. However, when eosinophils were coincubated with neutrophils and stimulated with IL-8, the TBM of eosinophils was significantly augmented. This augmented TBM of eosinophils was inhibited by a matrix metalloproteinase-9 inhibitor, a leukotriene B-4 receptor antagonist, platelet-activating factor antagonists, or an anti-TNF-alpha monoclonal antibodies. These results suggest that neutrophils migrated in response to IL-8 may lead eosinophils to accumulate in the airways of asthma and possibly aggravate this disease.
  • M Kushiya, K Saito, Kikuchi, I, T Kobayashi, K Hagiwara, M Kanazawa, M Nagata
    INTERNATIONAL ARCHIVES OF ALLERGY AND IMMUNOLOGY 140 17 - 22 1018-2438 2006 [Refereed][Not invited]
    Background: beta 2-Agonists, a representative class of bronchodilators used for asthma, have been shown to modulate some functions of eosinophils, including cell adhesion. Similarly, a leukotriene receptor antagonist (LTRA) may be beneficial in controlling inflammation in asthma, as cysteinyl leukotrienes (cysLTs) can cause accumulation or activation of eosinophils. Recent evidence suggests that the addition of an LTRA, but not a long-acting beta 2-agonist, to inhaled corticosteroid additionally reduces the number of eosinophils in sputum and blood from patients with asthma. The present study examined whether a beta(2)-agonist and an LTRA differentially modify eosinophil adhesion and activation induced by cysLTs and other activators. Methods: Eosinophils were isolated from blood of healthy donors and then incubated in the presence or absence of salbutamol (albuterol) or montelukast. Eosinophils were then exposed to leukotriene D-4 (LTD4) or another activator, and the generation of superoxide anion (O-2(-)) was evaluated by cytochrome C reduction assay. Eosinophil adhesion was examined by an eosinophil peroxidase assay. Results: Montelukast, but not salbutamol (both at 1 mu M), inhibited LTD4-induced (100 nM) eosinophil adhesion to recombinant human intercellular adhesion molecule 1. Both drugs similarly and partially inhibited the 100 pM interleukin-5-induced adhesive response of eosinophils to recombinant human intercellular adhesion molecule 1. Montelukast, but not salbutamol, blocked LTD4-induced eosinophil O-2(-) generation of eosinophils. Finally, neither salbutamol nor montelukast modified phorbol myristate acetate (11 ng/ml)-induced O-2(-) generation from eosinophils. Conclusion: These results confirm that LTD4 directly induces activation of eosinophils via the cysLT1 receptor. Furthermore, the results suggest that a beta(2)-agonist has no effect on eosinophil adhesion and activation induced by cysLTs. These results explain the differential effects of an LTRA and a beta(2)-agonist in the treatment of eosinophilic inflammation in asthma. Copyright (c) 2006 S. Karger AG, Basel.
  • Y Nagai, H Miyazawa, Huqun, T Tanaka, K Udagawa, M Kato, S Fukuyama, A Yokote, K Kobayashi, N Kanazawa, K Hagiwara
    CANCER RESEARCH 65 (16) 7276 - 7282 0008-5472 2005/08 [Refereed][Not invited]
    Lung cancer is one of the leading causes of the cancer death worldwide. Gefitinib is an inhibitor of the tyrosine kinase activity of the epidermal growth factor receptor (EGFR) and has been introduced in the treatment of advanced lung cancers. The responsiveness to gefitinib has been linked to the presence of EGFR mutations. Clinical samples contain many normal cells in addition to cancer cells. A method capable of detecting EGFR mutations in a large background of wild-type EGFR genes could provide a superior clinical test. We developed a rapid and sensitive detection system for EGFR mutations named the peptide nucleic acid-locked nucleic acid (PNA-LNA) PCR clamp that can detect EGFR mutations in the presence of 100-to 1,000-fold background of wild-type EGFR. We used this method to screen 30 non-small cell lung cancer cell lines established from Japanese patients. In addition to 11 cell lines that have mutations, we found 12 cell lines in which specific mutations are observed only in the subpopulation(s) of the cells. Genetic heterogeneity of EGFR suggests that the EGFR gene is unstable in established cancers and the heterogeneity may explain variable clinical responses of lung cancers to gefitinib.
  • H Xin, T Kikuchi, S Andarini, S Ohkouchi, T Suzuki, T Nukiwa, Huqun, K Hagiwara, T Honjo, Y Saijo
    EUROPEAN JOURNAL OF IMMUNOLOGY 35 (5) 1371 - 1380 0014-2980 2005/05 [Refereed][Not invited]
    The CX3C chemokine fractalkine (CX3CL1) exists as both a membrane-bound form promoting firm cell-cell adhesion and a soluble form chemoattracting leukocytes expressing its receptor CX3CR1. When adenoviral vector expressing mouse fractalkine (AdFKN) was transduced to the tumor cells, fractalkine was expressed as both membrane-bound form on the tumor cells and soluble form in the supernatant in vitro. Intratumoral injection of AdFKN (1x10(9)PFU/tumor) into C26 and B16F10 tumors resulted in marked reduction of tumor growth compared to control (C26: 86.5%, p< 0.001; B16F10: 85.5%,p< 0.001). Histological examination of tumor tissues revealed abundant infiltration of NK cells, dendritic cells, and CD8(+) T lymphocytes 3 and/or 6 days after treatment with AdFKN. Splenocytes from mice treated by AdFKN developed tumor-specific cytotoxic T cells, and thereby protected from rechallenging with parental tumor cells. Antitumor effects by AdFKN were completely abrogated in both NK cell-depleted mice and CD8(-/-) mice, and partially blocked in CD4(-/-) mice. These data indicated that fractalkine mediates antitumor effects by both NK cell-dependent and T cell-dependent mechanisms. This study suggests that fractalkine can be a suitable candidate for immunogene therapy of cancer because fractalkine induces both innate and adaptive immunity.
  • R Tazawa, E Hamano, T Arai, H Ohta, O Ishimoto, K Uchida, M Watanabe, J Saito, M Takeshita, Y Hirabayashi, Ishige, I, Y Eishi, K Hagiwara, M Ebina, Y Inoue, K Nakata, T Nukiwa
    AMERICAN JOURNAL OF RESPIRATORY AND CRITICAL CARE MEDICINE 171 (10) 1142 - 1149 1073-449X 2005/05 [Refereed][Not invited]
    The anti-granulocyte-macrophage colony-stimulating factor (GM-CSF) autoantibody is inferred to cause idiopathic pulmonary alveolar proteinosis (iPAP): the antibody neutralizes GM-CSF and thereby impairs differentiation of alveolar macrophages. Administration of GM-CSF improves respiratory function of patients with iPAP, as confirmed in this study using aerosolized GM-CSF. To elucidate its mechanism, we characterized bronchoalveolar lavage fluid and alveolar macrophages obtained from three patients with iPAP who were treated successfully with aerosolized GM-CSF. Cell number, expressions of surface mannose receptor and the transcription factor PU.1, and phagocytic ability of alveolar macrophages were all restored to control levels. With treatment, the neutralizing capacity of GM-CSF activity was reduced markedly, concomitant with the decreasing autoantibody levels. Interestingly, the amount of GMCSF autoantibody complex also decreased. In one case in which the complex was analyzed, the majority of GM-CSF binding the complex was endogenous protein, suggesting that the complex is removed immediately from the lung after treatment. Our study shows that GM-CSF administration engenders a decrease in the neutralizing capacity against the protein in the lungs. Thereby, it facilitates restoration of the normal function of alveolar macrophages.
  • T Shirai, S Miyagi, D Horiuchi, T Okuda-Katayanagi, M Nishimoto, M Muramatsu, Y Sakamoto, M Nagata, K Hagiwara, A Okuda
    JOURNAL OF BIOLOGICAL CHEMISTRY 280 (8) 7244 - 7252 0021-9258 2005/02 [Refereed][Not invited]
    The extraembryonic endoderm is derived from inner cell mass cells of the blastocyst during early mouse embryogenesis. Formation of the extraembryonic endoderm, which later contributes to the yolk sac, appears to be a prerequisite for subsequent differentiation of the inner cell mass. While embryonic stem cells can be induced to differentiate into extraembryonic endoderm cells in vitro, the molecular mechanisms underlying this process are poorly understood. We used a promoter trap approach to search for genes that are expressed in embryonic stem cells and are highly up-regulated during differentiation to the extraembryonic endoderm fate. We showed that fibronectin fits this expression profile. Moreover we identified an enhancer in the 12th intron of the fibronectin locus that recapitulated the endogenous pattern of fibronectin expression. This enhancer carries Sox protein-binding sequences, and our analysis demonstrated that Sox7 and Sox17, which are highly expressed in the extraembryonic endoderm, were involved in enhancer activity.
  • Suzuki T, Fukuhara T, Tanaka M, Nakamura A, Akiyama K, Sakakibara T, Koinuma D, Kikuchi T, Tazawa R, Maemondo M, Hagiwara K, Saijo Y, Nukiwa T
    Clinical cancer research : an official journal of the American Association for Cancer Research 11 (1) 58 - 66 1078-0432 2005/01 [Refereed][Not invited]
  • S Kikuchi, M Nagata, L Kikuchi, K Hagiwara, M Kanazawa
    INTERNATIONAL ARCHIVES OF ALLERGY AND IMMUNOLOGY 137 7 - 11 1018-2438 2005 [Refereed][Not invited]
    Background: Eosinophils are generally recognized as effector cells in asthma. Recently, neutrophils have been suggested to contribute to the development of chronic severe asthma. The mechanisms by which neutrophils contribute to the pathophysiology of asthma remain to be elucidated; however, neutrophils may affect either accumulation or functional status of eosinophils via the generation of inflammatory mediators. The objective of this study was to evaluate whether neutrophilic inflammation is associated with eosinophilic inflammation in severe asthma. Methods: Following the inhalation of hypertonic saline, induced sputum was obtained from 12 healthy controls, 10 mild persistent asthmatics who were treated with low-dose inhaled corticosteroids, and 8 severe persistent asthmatics who were treated with combinations of drugs including high-dose inhaled corticosteroids and oral prednisolone. Subsequently, differential inflammatory cell counts were evaluated. Results: The percentage of eosinophils in induced sputum was significantly higher in patients who showed airway neutrophilia. In severe persistent asthmatics, the percentage of neutrophils was significantly correlated with the percentage of eosinophils in induced sputum. Conclusions: The results of the present study suggest that accumulated neutrophils may contribute to the development of eosinophilic inflammation in severe persistent asthmatics who were treated with oral and high-dose inhaled corticosteroids. This effect may contribute to the eventual manifestation of airway inflammation in severe asthma. Copyright (C) 2005 S. Karger AG, Basel.
  • M Nagata, K Saito, Kikuchi, I, K Hagiwara, M Kanazawa
    INTERNATIONAL ARCHIVES OF ALLERGY AND IMMUNOLOGY 137 2 - 6 1018-2438 2005 [Refereed][Not invited]
    Background: Evidence shows that leukotriene receptor antagonist (LTRA) can cause a partial reduction of eosinophils in the asthmatic airway. Although cysteinyl leukotrienes (CysLTs) can regulate the development of eosinophilic inflammation, LTRA might modulate the eosinophilic response to other inflammatory molecules involved in allergic inflammation. Montelukast is an LTRA that inhibits eosinophil transendothelial migration (TEM) in response to platelet-activating factor (PAF). The present study evaluates whether pranlukast (an LTRA) modifies eosinophil TEM in response to chemoattractants including PAF and C-C chemokines. Methods: Eosinophils isolated from the blood of healthy individuals were incubated with or without pranlukast. We then evaluated eosinophil transmigration across human umbilical vein endothelial cells in response to LTD4, eotaxin, RANTES and PAR Results: Pranlukast did not modify the spontaneous transmigration of eosinophils (n = 5). As reported, eosinophil TEM was significantly augmented by 0.1 mu M LTD4 and this enhancement was blocked by 1 mu M pranlukast (p < 0.001; n = 6). On the other hand, pranlukast did not modify eosinophil transmigration in response to eotaxin, RANTES, or PAF (p > 0.1; n = 5). Conclusion: The inhibitory effect of pranlukast on eosinophil transmigration is highly specific for the CysLT1-dependent pathway. Copyright (C) 2005 S. Karger AG, Basel.
  • M Nagata, K Saito, Kikuchi, I, K Tabe, K Hagiwara, M Kanazawa, Y Sakamoto
    INTERNATIONAL ARCHIVES OF ALLERGY AND IMMUNOLOGY 134 21 - 24 1018-2438 2004 [Refereed][Not invited]
    Background: Eosinophil transendothelilal migration across vascular endothelial cells is an initial step of eosinophil accumulation in allergic inflammation. There is increasing evidence that specific immunotherapy (SIT) modulates the production of inflammatory molecules from mononuclear cells. Objective: The present study was undertaken to examine whether SIT modifies eosinophil transendothelial migration induced by the supernatants of antigen-stimulated mononuclear cells from atopic asthmatics. Methods: Dermatophagoides farinae (Df)-sensitive mild persistent asthmatics were divided into a SIT-treated group and a control group. Peripheral blood mononuclear cells (PBMC) were isolated before and after SIT using the rush protocol, and cultured for 96 h at 37 C in the presence or absence of Df antigen. Eosinophils were isolated from the blood of healthy subjects, and put on transwell filters coated with pulmonary microvascular endothelial cell monolayers stimulated with IL-4 plus TNF-alpha. The supernatants of PBMC were applied to the lower compartment and the transmigration of eosinophils was examined. Results: Df stimulation of PBMC resulted in an augmentation of eosinophil transendothelial migration. This enhancement was abrogated following SIT. In the control group, the antigen-induced effect on eosinophil transmigration did not show an interval change. Conclusion: SIT attenuates eosinophil transendothelial migration induced by antigen-stimulated mononuclear cells. Copyright (C) 2004 S. Karger AG, Basel.
  • T Fukushima, S Suzuki, M Mashiko, T Ohtake, Y Endo, Y Takebayashi, K Sekikawa, K Hagiwara, S Takenoshita
    ONCOGENE 22 (41) 6455 - 6457 0950-9232 2003/09 [Refereed][Not invited]
    BRAF is a serine/threonine kinase that receives a mitogenic signal from RAS and transmits it to the MAP kinase pathway. Recent studies have reported that mutations of the BRAF gene were detected with varying frequencies in several cancers, notably more than 60% in melanoma. We analysed mutations of BRAF and RAS genes in 100 cases of thyroid carcinoma to investigate genetic aberrations in the RAS/RAF/MEK/MAP kinase pathway. BRAF mutations were detected exclusively in papillary carcinomas ( 40 in 76 cases: 53%), and were exclusively V599E, a mutation frequently observed in other carcinomas. NRAS mutation was observed in six cases (6%), all in histological types other than papillary carcinoma, and was exclusively Q61R. No mutations were found in KRAS or HRAS. Our results suggest that BRAF mutations may play a critical role in the carcinogenesis of papillary carcinoma of the thyroid.
  • Huqun, Y Endo, H Xin, M Takahashi, T Nukiwa, K Hagiwara
    CANCER SCIENCE 94 (8) 718 - 724 1347-9032 2003/08 [Refereed][Not invited]
    p73, a close homolog of p53 tumor suppressor, induces growth arrest and apoptosis. However, its role in cancers is controversial because of the rarity of p73 mutations, lack of tumors in p73-knockout mice, and the presence of multiple isotypes, among which DeltaN isotypes inhibit the function of TA isotypes. We analyzed three naturally occurring p73 mutants found in lung cancer cell lines, NCl-H1155, DMS 92 and A427. NCI-H1155 is a cell line that has a p73 mutation [p73(G264W)] in the DNA-binding domain, as well as a p53 mutation [p53(R273H)], which is frequently found in human cancers and has a "gain-of-function" characteristic. p73alpha(G264W) not only lacks transactivation activity itself, but also suppressed the transactivation activity of the wild-type p73a in a dose-dependent manner, indicating that p73alpha(G264W) is a dominant-negative mutant. p73alpha(G264W) failed to suppress colony formation. We tested two other mutations, p73(Del418) in DMS 92 and p73(Del603) in A427. Both mutants retained similar levels of transactivation activity and suppression of colony formation to those of wild-type p73. The biological significance of these two mutations is unclear. In NCI-H1155 cells the coexistence of mutations that abrogate the normal functions of p73 and p53 may indicate that each mutation confers an additive growth advantage upon the cells.
  • K Usui, Y Saijo, K Narumi, S Koyama, M Maemondo, T Kikuchi, R Tazawa, K Hagiwara, Y Ishibashi, S Ohta, T Nukiwa
    ONCOGENE 22 (17) 2655 - 2663 0950-9232 2003/05 [Refereed][Not invited]
    Therapeutic modalities that overcome the antiapoptotic function of Bcl-2 that is often overexpressed in cancer cells are expected to be a novel strategy for cancer treatment. We previously reported that the leukemic cell death induced by an N-terminally truncated Bax (DeltaN Bax: corresponding to amino acid 112-192 of full-length Bax) was not blocked by Bcl-2 or Bcl-x(L) owing to the lack of the BH3 domain needed to interact with the antiapoptotic Bcl-2 family molecules. In this study, we used the Cre-loxP system that allowed us to propagate adenoviruses expressing DeltaN Bax, and investigated the effects of the DeltaN Bax gene transfer into A549 and NCI-H1299 nonsmall cell lung cancer cell tines. DeltaN Bax showed more cell-death-inducing activity in both cells than did the full-length Bax in vitro. It was found that the DeltaN Bax-induced cell death was not inhibited by the pan-caspase inhibitor z-VAD-fmk, suggesting that DeltaN Bax induces cell death through a caspase-independent mechanism. Intratumoral injection of adenoviruses expressing AN Bax into A549 tumors in Balb/c nude mice showed a significantly stronger suppression of tumor growth (74%) than full-length Bax (25%) compared to the control. Our results suggest that DeltaN Bax may provide a better alternative than currently used cytotoxic genes in cancer gene therapy trials.
  • Okohchi S, Tazawa R, Kimura Y, Miura R, Oshima M, Ebina M, Hagiwara K, Watanabe A, Nukiwa T
    Nihon Kokyuki Gakkai zasshi = the journal of the Japanese Respiratory Society 41 (4) 315 - 319 1343-3490 2003/04 [Refereed][Not invited]
    A 49-year-old man who had been treated with vitamin D since idiopathic hypoparathyroidism (IPH) was diagnosed at age 15 was referred to our hospital because of progressive wheezing. The same condition was also diagnosed in his sister at age 19. Chest radiography and computed tomography (CT) showed diffuse tracheal stenosis to a diameter of 6-8 mm. Laryngoscopy revealed no abnormality in the patient's epiglottis or vocal cords. No intact parathyroid hormone was detected in his sera but a brain CT scan showed calcification of the basal ganglia, which was compatible with IPH. Steroid therapy relieved his wheezing occasionally, but with little improvement of the narrowing of his trachea. He died of respiratory failure accompanied by lung abscess 15 weeks later. Autopsy revealed tracheal stenosis, thickening of the tracheal wall with marked fibrosis, and slight infiltration of inflammatory cells, which supported a diagnosis of relapsing polychondritis. This is the first report of relapsing polychondritis associated with IPH. Since both diseases are associated with autoimmune activity, this case may provide some insight into the pathogeneses of both diseases.
  • Kikuchi T, Hagiwara K, Honda Y, Watanabe A, Nukiwa T
    The Japanese journal of antibiotics 56 Suppl A 100 - 105 0368-2781 2003/04 [Refereed][Not invited]
  • A Nakamura, Y Mori, K Hagiwara, T Suzuki, T Sakakibara, T Kikuchi, T Igarashi, M Ebina, J Miyazaki, T Takai, T Nukiwa, T Nukiwa
    JOURNAL OF EXPERIMENTAL MEDICINE 197 (5) 669 - 674 0022-1007 2003/03 [Refereed][Not invited]
    Secretory leukoprotease inhibitor (SLPI) protects tissue against the destructive action of neutrophil elastase at the site of inflammation. Recent studies on new functions of SLPI have demonstrated that SLPI may play a larger role in innate immunity than merely as a protease inhibitor. To clarify the functions of SLPI in bacterial infections, we generated SLPI-deficient mice (SLPI-/- mice) and analyzed their response to experimental endotoxin shock induced by lipopolysaccharide (LPS). SLPI-/- mice showed a higher mortality from endotoxin shock than did wild type mice. This may be explained in part by our observation that SLPI-/- macrophages show higher interleukin 6 and high-mobility group (HMG)-1 production and nuclear factor kappaB activities after LPS treatment than do SLPI+/+ macrophages. SLPI also affects B cell function. SLPI-/- B cells show more proliferation and IgM production after LPS treatment than SLPI+/+ B cells. Our results suggest that SLPI attenuates excessive inflammatory responses and thus assures balanced functioning of innate immunity.
  • Hagiwara K, Kikuchi T, Endo Y, Huqun, Usui K, Takahashi M, Shibata N, Kusakabe T, Xin H, Hoshi S, Miki M, Inooka N, Tokue Y, Nukiwa T
    Journal of immunology (Baltimore, Md. : 1950) 170 (4) 1973 - 1979 0022-1767 2003/02 [Refereed][Not invited]
  • Kobayashi T, Miki M, Kikuchi T, Takahashi H, Hagiwara K, Watanabe A, Nukiwa T, Uchiyama B, Tateda K, Yamaguchi K
    Nihon Naika Gakkai zasshi. The Journal of the Japanese Society of Internal Medicine 91 (6) 1861 - 1863 0021-5384 2002/06 [Refereed][Not invited]
  • Tazawa R, Pradono P, Hagiwara K, Nukiwa T
    Nihon rinsho. Japanese journal of clinical medicine 60 Suppl 5 625 - 629 0047-1852 2002/05 [Refereed][Not invited]
  • T Kikuchi, K Hagiwara, Y Honda, K Gomi, T Kobayashi, H Takahashi, Y Tokue, A Watanabe, T Nukiwa
    JOURNAL OF ANTIMICROBIAL CHEMOTHERAPY 49 (5) 745 - 755 0305-7453 2002/05 [Refereed][Not invited]
    Erythromycin and other macrolides are effective for the treatment of chronic inflammatory airway diseases such as diffuse panbronchiolitis (DPB) and chronic sinusitis. The effect of macrolides in DPB is suggested to be anti-inflammatory rather than antibacterial. We investigated the effects of clarithromycin on interleukin-8 (IL-8) production using human peripheral monocytes and the human monocytic leukaemia cell line, THP-1. Bacterial extracts from Escherichia coli, Pseudomonas aeruginosa and Helicobacter pylori, as well as E. coli-derived lipopolysaccharide (LPS), induced IL-8 production. Clarithromycin suppressed this production in a dose-dependent manner in both monocytes and THP-1 cells (49.3-75.0% inhibition at 10 mg/L). A luciferase reporter gene assay with plasmids containing a serially deleted IL-8 promoter fragment showed that both the activator protein-1 (AP-1) and/or the nuclear factor-kappaB (NF-kappaB) binding sequences were responsible for the LPS and clarithromycin responsiveness of the IL-8 promoter. Consistently, in an electromobility shift assay, LPS increased the specific binding of both AP-1 and NF-kappaB, whereas clarithromycin suppressed it. Moreover, LPS and clarithromycin regulated three other promoters that have either the NF-kappaB or the AP-1 binding sequences: two synthetic (pAP-1-Luc and pNF-kappaB-Luc) and one naturally occurring (ELAM-Luc). Our results indicate that clarithromycin modified inflammation by sup-pressing IL-8 production and that clarithromycin may affect the expression of other genes through AP-1 and NF-kappaB. In addition to treatment of airway diseases, the anti-inflammatory effect of macrolides may be beneficial for the treatment of other inflammatory diseases such as chronic gastritis caused by H. pylori.
  • M Maemondo, K Narumi, Y Saijo, K Usui, M Tahara, R Tazawa, K Hagiwara, K Matsumoto, T Nakamura, T Nukiwa
    MOLECULAR THERAPY 5 (2) 177 - 185 1525-0016 2002/02 [Refereed][Not invited]
    Hepatocyte growth factor (HGF) affects tumor growth/invasion and tumor neovascularization. A proposed HGF antagonist, NK4 (an amino-terminal kringle-domain peptide of HGF), inhibits tumor growth/invasion through the competition of HGF binding to its receptor, c-Met, and acts as an angiogenesis inhibitor. To investigate the in vivo effect of NK4 gene transfer, we constructed an adenovirus vector expressing human NK4 (AdCMV.NK4). Human lung cancer cell lines (A549 and H358) infected in vitro with AdCMV.NK4 yielded NK4 protein without a change in the cell growth rate. In contrast, direct injection of AdCMV.NK4 (1 x 10(9) pfu, twice) into established subcutaneous tumors in BALB/c nu/nu mice resulted in suppression of the tumors by 64% for A549 or by 91% for H358 compared with controls (P < 0.02 or P < 0.01, respectively). Counting of the tumor vessels revealed suppressed vascularity by 57% in H358 tumors when using AdCMV.NK4 (P < 0.0001). Furthermore, systemic NK4 delivery by intraperitoneal injection of AdCMV.NK4 effectively suppressed both angiogenesis in the Matrigel assay (86% reduction, P < 0.032), subcutaneous tumor growth in vivo (by 65% for H358, P < 0.001), and hematogenous lung metastases without obvious side effects. These results indicate that NK4 elicits tumor-growth suppression in vivo through its anti-angiogenic activity and anti-HGF activity and that NK4 gene transfer can be an effective tool in the treatment of cancer.
  • Koinuma D, Miki M, Ebina M, Tahara M, Hagiwara K, Kondo T, Taguchi Y, Nukiwa T
    Internal medicine (Tokyo, Japan) 41 (1) 26 - 29 0918-2918 2002/01 [Refereed][Not invited]
  • P Pradono, R Tazawa, M Maemondo, M Tanaka, K Usui, Y Saijo, K Hagiwara, T Nukiwa
    CANCER RESEARCH 62 (1) 63 - 66 0008-5472 2002/01 [Refereed][Not invited]
    Cyclooxygenase, involved in tumor growth and angiogenesis, converts arachidonic acid to prostaglandin (PG)H-2, which is immediately converted to bioactive prostanoids including PGE(2), PGD(2), thromboxane (TX)A(2) and PGI(2). To test the hypothesis that changes in the prostanoid profile alter cancer growth, we transduced the retroviral vectors carrying TXA(2) synthase cDNA or PGI(2) synthase cDNA to colon-26 adenocarcinoma cells and subsequently inoculated each transformant to syngeneic BALB/c mice. Tumors derived from TXA(2) synthase transformants grew faster (280%, day 8, versus null-vector control; P < 0.05) and showed more abundant vasculature (204%, versus null-vector control; P < 0.01), whereas tumors from PGI(2) synthase transformants presented opposite effects. These effects by the transgenes were reversed by administration of specific inhibitors. These results suggest that the profile of downstream metabolites of cyclooxygenase in cancer cells can be a determinant for tumor development.


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