Teruo Iwata

Association Between Estrogen Receptor-{beta} Expression and Epidermal Growth Factor Receptor Mutation in the Postoperative Prognosis of Adenocarcinoma of the Lung

PURPOSE Adenocarcinoma of the lung unrelated to a smoking habit occurs more frequently in women than men, thus suggesting an association between female hormones and development of these tumors. The aim of this study was to elucidate the correlation between expression of estrogen receptor (ER) and clinicopathologic factors, including a mutation in the tyrosine kinase domain of epidermal growth factor receptor (EGFR), and prognosis in adenocarcinoma of the lung. PATIENTS AND METHODS This study evaluated 447 resected primary lung adenocarcinoma specimens. The expression of ERalpha and ERbeta was evaluated with an immunohistochemical method. The EGFR mutation was evaluated with polymerase chain reaction. RESULTS A strong cytoplasmic expression of ERalpha and nuclear expression of ERbeta were detected in 49.4% and 48.5% of all patients, respectively. A strong nuclear expression of ERbeta was independently associated with the EGFR mutations (odds ratio = 2.947; 95% CI, 1.97 to 4.57; P < .001) and good differentiation (odds ratio = 1.84; 95% CI, 1.21 to 2.80; P = .004) and was correlated with an increasing disease-free survival in patients with EGFR mutations (hazard ratio = 2.18; 95% CI, 1.18 to 4.06; P = .014). However, no prognostic significance was identified in patients without EGFR mutations. No clinicopathologic and/or prognostic significance of a strong expression of cytoplasmic ERalpha was found. CONCLUSION A strong nuclear expression of ERbeta correlates with EGFR mutations, and its favorable prognostic significance was influenced by the EGFR mutations in adenocarcinoma of the lung.

Expression of estrogen receptor beta predicts a clinical response and longer progression-free survival after treatment with EGFR-TKI for adenocarcinoma of the lung.

PURPOSE Epidermal growth factor receptor (EGFR)-tyrosine kinase inhibitor (EGFR-TKI) demonstrates a dramatic clinical response for the lung adenocarcinoma patients harboring a somatic mutation of EGFR. Such EGFR mutations are frequently found in adenocarcinoma with a strong expression of estrogen receptor (ER) beta, which has been shown to correlate with a favorable prognosis for the patients with EGFR mutations. The aim of this study is to elucidate the correlation between expression of ER beta and the therapeutic effect of EGFR-TKI in adenocarcinoma of the lung. PATIENTS AND METHODS Forty-three patients who were treated with EGFR-TKI for adenocarcinoma of the lung were evaluated. The expression of ER beta and the EGFR mutation were evaluated by immunohistochemistry and the polymerase chain reaction, respectively. Patients divided into two groups by the nuclear expression of ER beta. The clinical response and survival data were compared between the two groups. RESULT Strong (S) and weak (W) expression of ER beta was observed in 21 and 22 patients, respectively. EGFR mutations were detected in 30 (69.8%) cases. The S group had more frequent EGFR mutations than the W group (85.7%, 54.5%, p=0.045). The S group had better response rate (p=0.006) and longer progression-free survival (PFS; p=0.001) than the W group. Even in a limited analysis in the patients with EGFR mutations, the S group had tended to have a better response rate (77.8%, 41.7%, p=0.063), and significant longer PFS (p=0.012) than the W group. CONCLUSION A strong expression of ER beta predicts a good clinical outcome for patients with adenocarcinoma of the lung after treatment with EGFR-TKI. This suggests that the expression status of ER beta can be a candidate surrogate marker for EGFR-TKI treatment of patients with adenocarcinoma of the lung. Further investigation will be necessary to identify biomarkers using a larger cohort of patients in a prospective study.

TS expression predicts postoperative recurrence in adenocarcinoma of the lung

BACKGROUND Not all patients with lung cancer require postoperative adjuvant chemotherapy after a complete resection. However, no useful markers for either selecting appropriate candidates or for predicting clinical recurrence exist. METHODS Tumor specimens were collected from 183 consecutive patients who underwent a complete resection for lung adenocarcinoma from 2003 to 2007 in our department. We analyzed the thymidylate synthase (TS) and dihydrofolate reductase (DHFR) expressions in the primary lung adenocarcinoma by immunohistochemisty. RESULTS The strong expression of TS and DHFR was identified in 39 (21.3%) and 120 (65.6%) patients, respectively. The strong TS expression was identified in 11 (39.3%) of 28 patients and 28 (18.1%) of 155 patients in patients with and without recurrence, respectively (p=0.012). The strong DHFR expression was also identified in 23 (82.1%) and 97 (62.6%) of the patients with and without recurrence, respectively (p=0.045). Logistic regression models indicated the strong TS expression to be an independent factor for tumor recurrence. The strong TS and DHFR expression was associated with a poorer disease-free survival (DFS) according to the survival analysis. A multivariate analysis demonstrated the strong TS expression to be independently associated with an increased risk for poor DFS. CONCLUSIONS The strong TS expression may be a useful marker for predicting postoperative recurrence in patients with lung adenocarcinoma following surgery.

Clinical significance of the frequency of regulatory T cells in regional lymph node lymphocytes as a prognostic factor for non-small-cell lung cancer.

BACKGROUND Regulatory T cells (Tregs) are potent immunosuppressive cells that play a crucial role in tumor immune escape. The purpose of the present study was to evaluate the prognostic significance of the frequency of CD4+CD25+Foxp3+ Tregs in the regional lymph node lymphocytes (RLNL) and peripheral blood lymphocytes (PBL) in patients who underwent surgical resection of non-small cell lung cancer (NSCLC). METHODS The RLNL and PBL in 158 NSCLC patients who underwent complete surgical resection were collected at the time of surgery. The proportions of CD4+CD25+Foxp3+ cells in the RLNL and PBL were determined by flow cytometry. RESULTS The average proportions of Tregs in the RLNL and PBL were 1.28% and 0.76%, respectively. The proportion of Tregs in the RLNL was significantly higher than that in the PBL (p < 0.0001). The 5-year overall survival rates of the patients according to the proportion of Tregs in the RLNL were 84.4% and 63.5% in the lower and higher groups, respectively. A significant difference was observed in the survival rate between the higher and lower groups (p = 0.0056). Among the patients with stage I disease, the 5-year survival rate (91.4%) was significantly higher in patients with the lower proportion of Tregs in RLNL that in the higher group (72.1%) (p = 0.0147). CONCLUSIONS The higher proportion of Tregs in the RLNL was a significant unfavorable prognostic factor, even in patients with node-negative NSCLC. The information about the proportion of Tregs in the RLNL might improve the discriminatory power for assessing the risk of the recurrence of NSCLC.

Increased cytochrome P450 and aryl hydrocarbon receptor in bronchial epithelium of heavy smokers with non-small cell lung carcinoma carries a poor prognosis.

Smoking induces mutations via the formation of DNA-adducts in the bronchial and alveolar epithelium and contributes to the development of lung cancer. Benz(a)pyrene and nitrosamine, typical carcinogens in cigarette smoke, undergo metabolic activation by the phase I enzymes, such as cytochrome P450 (CYP) 1A1, CYP2A6 and CYP2E1. The transcriptional regulation of these phase I enzymes is regulated by arylhydrocarbon receptor (AH-R) which binds many well-known carcinogens. To identify a cause and effect relationship, the expression of cytochrome CYP and AH-R in the bronchial epithelium was correlated with the history of cigarette smoking in patients with non-small cell lung carcinoma (NSCLC). Although CYP3A+ cells were absent in the bronchial epithelium of all patients, there were many CYP2E1+ cells in heavy (>1000 cigarette/day x year) smokers (38.5%). In contra-distinction, there was significantly less number of CYP2E1+ cells in light (less than 1000 cigarette/day x year) smokers (15.6%) or non-smokers (10.0%). Similarly, there were more CYP1A1+ (19.2%) and CYP2A6+ cells in heavy (65.4%) smokers as compared to non-smokers. The number of AH-R+ cells was also significantly higher in cases with p53 mutation (62.5%) than those without (12.2%) mutation. Since in patients with early NSCLC, CYP positivity showed a close correlation with a poor survival (p less than 0.01), expression of CYP in bronchial epithelium has a prognostic potential.

Detection of EGFR and K-ras mutations for diagnosis of multiple lung adenocarcinomas.

The incidence of multiple primary lung adenocarcinoma (MPLA) is increasing, and it is important to distinguish MPLA from intrapulmonary metastasis (IPM) in order to determine the therapeutic strategy. However, there is no reliable method to differentiate between the two. The purpose of this study was to distinguish MPLA from IPM based on the gene status of EGFR and K-ras and the morphological Noguchi classification system. Sixty-eight tumors from 34 cases of clinical MPLA were evaluated. Of them, 11 cases (32.4%) were diagnosed as biological MPLA (bMPLA) by EGFR/K-ras mutation analyses, and 12 cases (35.3%) by morphological analysis. In all, 23 of the 34 cases (67.6%) were diagnosed as bMPLA. The remaining 11 cases were diagnosed as biological IPM (bIPM). The 5-year survival rates of bMPLA and bIPM were 90.9% and 63.6%, respectively (p=0.04). These findings suggest that the combination method including gene mutation and morphological analysis can guide treatment decisions and that there is a need for systemic chemotherapy, and surveillance monitoring.

A Successful Case of Tracheal Segmental Resection and Reconstruction for Cicatricial Tracheal Stenosis

A 45-year-old man had consciousness disorder due to fall trauma had received ventilation support and tracheostomy. Two months later, the tracheostomy tube was removed. One year later, he suffered from severe cicatricial tracheal stenosis. Under a diagnosis of post-intubation tracheal stenosis, he underwent circumferential resection and end-to-end anastomosis of the trachea. The central part of the resected trachea of 3 cartilage rings showed a stenosis like a pin hole. The post operative course was uneventful, and there was no stenosis or sutural insufficiency on examination by bronchoscopy. Tracheal resection and reconstruction is rare but effective for refractory tracheal stenosis.

Abstract 5103: Development of detecting systems for circulating tumor cells in malignant pleural mesothelioma patients.

Proceedings: AACR 104th Annual Meeting 2013; Apr 6-10, 2013; Washington, DC Background. To isolate and quantify of CTCs, the CellSearch system is the most commonly used techniques, approved by the FDA. But in this system, EpCAM negative CTCs are excluded. To detect CTCs in malignant pleural mesothelioma patients, we have developed two novel approaches, CTC-Chip and immunomagnetic beads which could be labeled with any type of antibodies. Methods. To evaluate the accuracy of the number of CTCs, we used the peripheral blood samples collected from healthy donors, spiked definite number of tumor cell lines, ACC-MESO1 and ACC-MESO4. Both tumor cell lines (obtained from ATCC) were derived from malingnant pleural mesothelioma, positive of mesothelin and podoplanin. The recovery rate was evaluated with two methods, the CTC-Chip and immunomagnetic beads, coated with antibodies specific for mesothelin and podoplanin. In addition, an 8 ml sample of peripheral blood was collected from a patient with malignant pleural mesothelioma, and was served for the immunomagnetic beads. To identify the captured cells as tumor cells, immunocytochemistry was performed using with the cocktail of two human cytokeratin-specific antibodies. Results. To confirm the efficacy of tumor isolation by two methods, we used tumor cell suspicion as samples. In CTC-Chip coated with anti-mesothelin and podoplanin, the recovery rate of captured ACC-MESO1 was 99%, 97%, respectively. The recovery efficiency of ACC-MESO1-spiked blood samples using CTC-Chip coated with anti-mesothelin and podoplanin, was 45%, 80%, respectively. In immunomagnetic beads coated with anti-mesothelin and podoplanin, the recovery rate of captured ACC-MESO4 was 80%, 25%, respectively. The recovery efficiency of ACC-MESO4-spiked blood samples using immunomagnetic beads coated with anti-mesothelin and podoplanin, was 33%, 15%, respectively. The CTCs from a patient was detected using with immunomagnetic beads coated with anti-mesothelin antibody and CTC count was 440 per 8 ml of peripheral blood. Conclusions. Both CTC-Chip and immunomagnetic beads could capture CTCs, using antibodies specific for mesothelin and podoplanin. Detection of CTCs from a patient with malignant pleural mesothelioma was performed by labeling of mesothelin-coated immunomagnetic beads. But the recovery rate of tumor cells in spiked samples were less than 30%. The step of deletion of red blood cells is necessary to be improved. Citation Format: Chikaishi Yasuhiro, Tomoko So, Soichi Oka, Masaru Takenaka, Makoto Nakagawa, Hidehiko Shimokawa, Teruo Iwata, Yoshika Nagata, Hidetaka Uramoto, Takeshi Hanagiri, Takeshi Ohnaga, Fumihiro Tanaka. Development of detecting systems for circulating tumor cells in malignant pleural mesothelioma patients. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 5103. doi:10.1158/1538-7445.AM2013-5103

Invasive Lobular Carcinoma of the Breast with Wide Spread Intestinal Metastases

症例は50歳の女性で, 両側乳腺浸潤性小葉癌の術後7年目に, 右側腹部圧痛, 便秘を訴え, 精査にて上行結腸の全周性狭窄, 胃のびらん性病変を認めた. いずれの病変も生検では低分化腺癌であり, 胃の生検では印環細胞を認めた. この消化管腫瘍の組織像は, 乳癌の組織像と類似し, 免疫染色でエストロゲンレセプターおよびプロゲステロンレセプター陽性であり, 乳癌の大腸転移・胃転移が疑われた. イレウス症状出現し, 結腸右半切除術を施行した. 大腸病変は粘膜下組織を中心に広がり, 乳癌と同様の組織所見であった. 乳腺浸潤性小葉癌では, 消化管への転移がありうるという特殊な転移様式を念頭においた全身検索が必要である. また, 浸潤性小葉癌で粘液産生の強い例では, 印環細胞の様相を呈すため, 胃転移症例では, 原発性胃癌との鑑別が必要である.