Hirotoshi Akita

A Phase III Comparison of Etoposide/Cisplatin with or without Added Ifosfamide in Small-Cell Lung Cancer

A total of 92 patients with small-cell lung cancer (SCLC) were randomized to receive cisplatin (80 mg/m2, day 1)/etoposide (100 mg/m2, days 1, 3, 5) (PE) or cisplatin (80 mg/m2, day 1)/etoposide (100 mg/m2, days 1, 3, 5)/ifosfamide (2 g/m2, days 1, 2, 3) (PEI) combination chemotherapy. After 2 courses of chemotherapy, patients with limited disease (LD) received chest irradiation of 40-50 Gy. Of the 89 patients who could be wholly evaluated, the overall response rate was 78% for PE and 74% for PEI therapy (NS). For all patients the complete response (CR) rates were 14 versus 21%, respectively, and 22 versus 30% for LD. However, the median survival times for all patients were 55 weeks for PE therapy versus 56 weeks for PEI therapy (NS). The 2-year survival rates were 15 and 17%, respectively, for all patients (NS). There was no difference in the duration of response between PE and PEI therapy in cases with CR or partial response. However, severe leukopenia (< 2,000/mm3) occurred more often after PEI (73%) than after PE (44%) therapy (p < 0.05). These results suggest that PEI is not superior to PE chemotherapy in SCLC. The use of ifosfamide in multimodality treatment regimens needs to be studied further.

Marked reduction of type I keratin (K14) in cisplatin-resistant human lung squamous-carcinoma cell lines

We have established two cisplatin-resistant human lung squamous-carcinoma cell lines, PC10-B3 and PC10-E5, from their original cell line PC10. To discover which proteins are associated with cisplatin resistance, we carried out a two-dimensional gel electrophoresis to analyze differences in protein alteration between PC10, PC10-B3 and PC10-E5. A protein spot M(r) 50 kDa, pI5.3, was reduced markedly and a spot M(r) 50 kDa, pI4.9 was increased when PC10-B3 and PC10-E5 were compared with PC10. A spot M(r) 58 kDa, pI5.8 newly appeared only in PC10-E5. Cell fractionation showed that the M(r) 50 kDa, pI5.3 (p50-5.3) and the M(r) 50 kDa, pI4.9 fell within the nuclear fraction, while the M(r) 58 kDa, pI5.8 was found among the cytosol and microsomal fractions. Microsequencing after in situ digestion of the dramatically reduced spot p50-5.3 revealed that it was identical to 50 kDa, type I keratin (K14). Moreover, a retinoic acid-mediated K14 reduction was concomitant with a 4.0-fold increase in cisplatin resistance in PC10. Our report is the first to suggest the possible association of marked K14 reduction and cisplatin resistance in PC10-B3 and PC10-E5.

Abstract 5162: Effects of combined epigenetic therapy with histone methyltransferase EZH2 inhibitor 3-deazaneplanocin and histone deacetylase inhibitor SAHA on non-small cell lung cancer cells

Proceedings: AACR Annual Meeting 2014; April 5-9, 2014; San Diego, CA [Background] EZH2, a polycomb group protein, has histone methyltransferase activity, and is involved in malignant transformation of several cancers. We have previously shown that an EZH2 inhibitor, DZNep, inhibits growth of non-small cell lung cancer(NSCLC) cell lines via G1 arrest and apoptosis. Recently, cotreatment with DZNep and an histone deacetylase (HDAC) inhibitor has been shown to induce apoptosis synergistically in AML, ovarian cancers and renal cancers. Therefore, we determined the combined effect of DZNep and an HDAC inhibitor, SAHA, on NSCLC cells. [Methods] We evaluated the effect of combined therapy with DZNep and SAHA against four human NSCLC cell lines_H1299, H1975, A549 and PC-3. Cell proliferation was measured by MTT assay. Percentage of apoptotic cells was measured using Annexin V-FITC with a flow cytometer. Western blot analysis was performed on total cell lysates. For in vivo assay, H1975 cells were subcutaneously implanted into the flank of BALB/cAJcl nu/nu nude mice. When the average tumor volume reached approximately 50∼100mm3, the following treatments were intraperitoneally administered twice per week for 6 weeks; vehicle alone (5% DM SO), DZNep (4mg/kg), SAHA (40mg/kg), and DZNep (4mg/kg) plus SAHA (40mg/kg). [Results] Co-treatment with DZNep and SAHA inhibited cell proliferation synergistically, and reduced EZH2 expression and histone H3 lysine 27 trimethylation more effectively compared with each agent alone. The co-treatment greatly induced accumulation of p27 Kip1 and decrease in cyclin A expression. Flow cytometry analysis demonstrated that the apoptotic fraction was increased in an additive or synergistic manner by the combination therapy. These effects were more evident in H1975 and PC-3 cells with EGFR mutation, in which the co-treatment markedly reduced phosphorylation of EGFR, AKT and ERK1/2. The combined therapy reduced the levels of β-catenin, and its downstream pro-proliferative targets cyclin D1 and EGFR. The co-treatment with DZNep and SAHA also caused significantly greater inhibition of tumor growth of H1975 xenografts than each agent alone in nude mice without visible toxicity. [Conclusion] Combined epigenetic therapy with an EZH2 inhibitor and an HDAC inhibitor may represent an effective approach for NSCLCs. Citation Format: Taichi Takashina, Ichiro Kinoshita, Junko Kikuchi, Yasushi Shimizu, Jun Sakakibara Konishi, Satoshi Oizumi, Masaharu Nishimura, Hirotoshi Dosaka Akita. Effects of combined epigenetic therapy with histone methyltransferase EZH2 inhibitor 3-deazaneplanocin and histone deacetylase inhibitor SAHA on non-small cell lung cancer cells. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 5162. doi:10.1158/1538-7445.AM2014-5162

Late Onset of Non-islet Cell Tumor Hypoglycemia Managed via Multidisciplinary Treatment in a Patient with a Solitary Fibrous Tumor

Solitary fibrous tumor (SFT) is a rare subtype of soft tissue sarcoma (STS). We herein describe a case of late onset of non-islet cell tumor hypoglycemia (NICTH) that was managed via multidisciplinary treatment in a patient with SFT. A 67-year-old man previously diagnosed with SFT 4 years prior to this presentation and treated with several rounds of surgery, presented with massive tumors. Eighteen months following his prescribed chemotherapy, the patient developed hypoglycemia. He was diagnosed with NICTH, after confirming the presence of high molecular weight insulin-like growth factor-2. This case suggests that paraneoplastic syndrome can occur even in cases of rare cancers, such as STS.

1190PA RANDOMIZED PHASE II TRIAL OF CISPLATIN PLUS GEMCITABINE VERSUS CARBOPLATIN PLUS GEMCITABINE IN PATIENTS WITH COMPLETELY RESECTED NON-SMALL CELL LUNG CANCER: HOKKAIDO LUNG CANCER CLINICAL STUDY GROUP TRIAL (HOT0703)

ABSTRACT Aim: The combination of platinum and gemcitabine (GEM) (P/G) is one of the most beneficial regimens in advanced non-small cell lung cancer (NSCLC). This study aimed to evaluate the efficacy and safety of P/G in adjuvant chemotherapy of postoperative NSCLC and select the P/G arm for a phase III trial. Methods: Patients with postoperative stage IB-IIIA NSCLC were randomly assigned to either cisplatin (CDDP) 40 mg/m2 on days 1 and 8 plus GEM 1000 mg/m2 on days 1 and 8 (GP arm) or carboplatin (CBDCA) AUC 5 on day 8 plus GEM 1000 mg/m2 on days 1 and 8 (GC arm) every 3 weeks for 4 cycles. The primary endpoint was 2-year disease-free survival (DFS); secondary endpoints were safety, feasibility, overall survival (OS), and biomarker analyses. Results: 102 patients were randomized (stages: 22% IB, 36% II, 42% IIIA; histology: 74% adenocarcinoma, 26% non-adenocarcinoma). Thirty-seven (73%) of 51 patients in each arm completed 4 cycles. Compliance was greater with GP than GC (dose intensity 87.6% vs. 77.9%, respectively; P Conclusions: Both P/G combination regimens were feasible and well-tolerated and, thus, may be potential options for adjuvant treatment in postoperative NSCLC. Although there were no significant differences in DFS between the two regimens, the data presented favor adopting GC for further evaluation. Disclosure: All authors have declared no conflicts of interest.

1465PUPDATED DATA ON CLINICAL AND MOLECULAR PROFILE OF SURGICALLY RESECTED SMALL CELL LUNG CANCER: INTERGROUP STUDY WITH FIGHT002 AND HOT1301

ABSTRACT Aim: NCCN and Japanese guidelines suggest surgery for patients with c-stage I small-cell lung cancer (SCLC), and ESMO guidelines recommend surgery for patients with c-stage II (T1,2 N0,1). However, the clinical impact of surgery with other variables on patients with early stage SCLC has yet to be determined. Therefore, clarification of the clinical and molecular profile of surgically resected SCLC is required. We expanded the number of patients and updated the clinical data which had been presented at ASCO 2014 (abstract #7590). Methods: We reviewed the clinical courses of 156 patients with SCLC who had undergone surgery at 17 institutes from January 2003 to January 2013. 125 formalin-fixed paraffin-embedded tissue samples were subjected to immunohistochemistry using 8 antibodies and to next-generation sequencing (NGS) systems using MiSeq and TruSight Tumor Sequencing Panel (Illumina) loading 26 genes. (UMIN registration No. 000010116 /10117). Results: Median relapse-free survival (RFS) and overall survival (OS) were 15.6 (95%CI: 6.8-24.5), and 33.3 (20.9-45.8) months, respectively. Multivariate analysis revealed that OS was longer in patients without history of malignancy (HR: 0.459, 95%CI: 0.248-0.847, p=0.013), with preoperative diagnosis (HR: 0.529, 95%CI: 0.293-0.953, p=0.034), and with c-stage II and under (HR: 0.117, 95%CI: 0.047-0.288, p Conclusions: These results support the ESMO guidelines for the management of c-stage II small-cell lung cancer, and indicate that history of malignancy might be a major decisive factor for surgery. The results of immunohistochemistry assist us in gaining a better understandingof the biology of SCLC. Disclosure: All authors have declared no conflicts of interest.

Abstract P4-04-08: A Potential non-viral vector to transfect dendritic cell and thereby MHC-Class I antigen presentation might be a potential use in carcinoma

To date almost no remarkable potential non-viral vector was developed to transfect promisingly primary dendritic cell like Bone Marrow Derived Dendritic Cell (BMDC) so that it might generate antigen presentation while epitopic-plasmid was delivered. Here we have introduced a potential non-viral vector named Stearyl-KALA MEND (100–130 nm, zeta potential 35–40 mV), which expressed to date the highest transgene expression while firefly luciferase was introduced (sub-cloned), and thereby promising antigen presentation in vitro while OVA plasmid containing MHC Class-I restricted SIINFEKL epitope was introduced and co-cultured with B3Z T-cell hybridoma. On the basis of these results, we used this vector to transfect BMDC and harvested the DC to chase the corresponding EG7-OVA induced tumor ex vivo. Later we further immunized mice directly with STR-KALA MEND containing OVA plasmid and challenged against the tumor (EG7-OVA induced). In this in vivo study we found also a significant antitumor activity. To evaluate the promptness of our vector we further sub-cloned Mart-1 gene, in our sub-cloned plasmid vector and immunized the mice as before. Thereafter inserting B16F10 melanoma cells to the immunized mice we found also a significant antitumor activity after 24 days of inoculation. Thus the vector, STR-KALA MEND might be a future potential use as DNA vaccine in anti-tumor methodology. Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P4-04-08.

Abstract 4118: Expression of CD133 as a prognostic marker of non-small cell lung cancers

[Background] CD133 is a membrance glycoprotein with five transmembrance loops. Previous reports have suggested that CD133-positive subpopulation of multipotent cells with extensive proliferative and self-renewal abilities has biological features as a cancer stem cell. In addition, it has been reported that the presence of CD133-positive cells was associated with a significantly poorer prognosis compared with the CD133-negative cells in some solid tumors, including colon cancer, hepatocellular carcinoma and glioma. However, little is known about the relationship between the expression of CD133 and clinical and clinicopathological characteristics in lung cancer. [Aim and Method] We conducted immunohistochemical assessment of 161 surgically resected non-small cell lung cancers (NSCLCs) at Hokkaido University Hospital between 1982 and 1994 to evaluate the correlation between CD133 expression and various features, including clinical and clinicopathological characteristics. [Results] Normal alveoli were used as negative controls and Bowman9s capsule basement membrane as positive controls. CD133 expression was significantly correlated with advanced p-Stage (p=0.040). The Kaplan-Meier method demonstrated that NSCLC patients with CD133 expression in tumor cells showed poor prognosis compared to those without CD133 expression in p-Stage II, III and IV (p=0.0098). CD133 expression alone was an independent factor for unfavorable prognostic in those patients (Hazard Ratio=3.157, p=0.015). [Conclusion] CD133 expression was correlated with p-Stage and was an independent factor for unfavorable prognosis in patients with p-Stage II, III and IV NSCLCs. CD133 expression may be a novel prognostic marker for NSCLCs. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 4118. doi:10.1158/1538-7445.AM2011-4118

A Case of a Small Lesion of Squamous Cell Carcinoma of the Peripheral Lung Diagnosed by Positive Findings on Sputum Cytologic Examination

背景. 喀痰細胞診陽性から発見された肺末梢発生の小型扁平上皮癌の1例を報告する. 症例. 77歳男性. 肺癌検診の喀疾細胞診で陽性と判定され精査を施行するも異常を認めなかった. 2年後, 胸部X線写真, 胸部CTにて右S3に径12mmの小型腫瘤影を認めた. 気管支鏡検査にて可視範囲に異常所見を認めず, 全支擦過細胞診を施行するも悪性所見は認めなかった. 右S3の腫瘤は6次分岐以降の末梢に存在するものと判断し, CTガイド下経皮生検を施行し, 扁平上皮癌と診断され, 60Gy/8回の定位放射線療法を施行した. 治療後約4年経過したが, 癌の再発は認めず, 喀痰細胞診も陰性を持続している. 結論. 肺末梢発生の小型扁平上皮癌が喀疾細胞診陽性の原因と考えられた.

Clinical Features of Primary Lung Cancer in Cases with Previous Lung Diseases.

1983年より1998年までに当科に入院した原発性肺癌755例のうち, 先行肺疾患それぞれの合併率と先行肺疾患合併肺癌の臨床的特徴を検討した.最も合併頻度が高かったのは肺結核で125例 (16.6%), 次いで肺気腫81例 (10.7%), 問質性肺炎30例 (4.0%), 気管支喘息14例 (1.9%), 珪肺11例 (1.5%) の順であった.肺気腫は合併率が年々急激に増加したが, 呼吸機能には変化はなかった.肺気腫, 間質性肺炎, 肺結核の合併は男性に多く (P<0.01), 肺気腫と肺結核の合併は高齢者, 重喫煙者に多く認められた (P<0.01).肺気腫, 間質性肺炎, 珪肺合併例で, それぞれの非合併例と比較して扁平上皮癌の比率が高く, 気管支喘息合併例では腺癌が多く認められたが, 肺気腫合併例 (P<0.01) を除き, 統計学的有意差は無かった.間質性肺炎合併例ではStage IIIA以上の進行した症例が多く (P=0.015), 化学療法単独と, 支持療法を受けた症例が多かった.以上より, 先行肺疾患合併原発性肺癌はそれぞれ特徴を持つことが確認され, 今後の原発性肺癌の病因解明にも有用な情報となることが期待される