-
Kunitoshi Shigeyasu,
Shunsuke Kagawa, Futoshi Uno,
Masahiko Nishizaki,
Hiroyuki Kishimoto,
Akira Gochi,
Toshikazu Kimura,
Takaomi Takahata,
Yasuyuki Nonaka,
Motoki Ninomiya,
Toshiyoshi Fujiwara
[show abstract]
[hide abstract]
ABSTRACT: PURPOSE: Peritoneal dissemination is the most frequent and life-threatening mode of metastasis and recurrence in patients with gastric cancer. A multicenter phase II study was designed to evaluate the efficacy and tolerability of S-1 and docetaxel combination chemotherapy regimen for the treatment of advanced or recurrent gastric cancer patients with peritoneal dissemination. METHODS: Nineteen patients with histologically confirmed unresectable or recurrent gastric cancer with peritoneal dissemination were enrolled. Oral S-1 at 80 mg/m(2)/day was administered twice daily for 2 weeks, followed by 1 drug-free week. Docetaxel infusion at 40 mg/m(2) was performed on day 1, simultaneous with S-1 administration. The primary endpoints were overall survival (OS) and time to progression (TTP). The secondary endpoints were the response rates and safety status. RESULTS: Patients received a median of 4 cycles of the S-1 and docetaxel regimen (range 1-43). The disease control rate was 73.7 % (14/19). Median overall survival was 459 days (15.3 months), while median time to progression was 212 days (7.1 months). Neutropenia was the most common type of toxicity (n = 7, 36.8 %). CONCLUSIONS: Combination chemotherapy with S-1 and docetaxel is a tolerable and effective treatment for advanced or recurrent gastric cancer patients with peritoneal dissemination.
Cancer Chemotherapy and Pharmacology 01/2013; · 2.83 Impact Factor
-
Joe Hasei,
Tsuyoshi Sasaki,
Hiroshi Tazawa,
Shuhei Osaki,
Yasuaki Yamakawa,
Toshiyuki Kunisada,
Aki Yoshida,
Yuuri Hashimoto,
Teppei Onishi, Futoshi Uno,
Shunsuke Kagawa,
Yasuo Urata,
Toshifumi Ozaki,
Toshiyoshi Fujiwara
[show abstract]
[hide abstract]
ABSTRACT: Tumor suppressor p53 is a multifunctional transcription factor that regulates diverse cell fates, including apoptosis and autophagy in tumor biology. p53 overexpression enhances the antitumor activity of oncolytic adenoviruses; however, the molecular mechanism of this occurrence remains unclear. We previously developed a tumor-specific replication-competent oncolytic adenovirus, OBP-301, that kills human osteosarcoma cells, but some human osteosarcoma cells were OBP-301-resistant. In this study, we investigated the antitumor activity of a p53-expressing oncolytic adenovirus, OBP-702, and the molecular mechanism of the p53-mediated cell death pathway in OBP-301-resistant human osteosarcoma cells. The cytopathic activity of OBP-702 was examined in OBP-301-sensitive (U2OS and HOS) and OBP-301-resistant (SaOS-2 and MNNG/HOS) human osteosarcoma cells. The molecular mechanism in the OBP-702-mediated induction of two cell death pathways, apoptosis and autophagy, was investigated in OBP-301-resistant osteosarcoma cells. The antitumor effect of OBP-702 was further assessed using an orthotopic OBP-301-resistant MNNG/HOS osteosarcoma xenograft tumor model. OBP-702 suppressed the viability of OBP-301-sensitive and OBP-301-resistant osteosarcoma cells more efficiently than OBP-301 or a replication-deficient p53-expressing adenovirus (Ad-p53). OBP-702 induced more profound apoptosis and autophagy when compared with OBP-301 or Ad-p53. E1A-mediated miR-93/106b upregulation induced p21 suppression, leading to p53-mediated apoptosis and autophagy in OBP-702-infected cells. p53 overexpression enhanced adenovirus-mediated autophagy through activation of damage-regulated autophagy modulator (DRAM). Moreover, OBP-702 suppressed tumor growth in an orthotopic OBP-301-resistant MNNG/HOS xenograft tumor model. These results suggest that OBP-702-mediated p53 transactivation is a promising antitumor strategy to induce dual apoptotic and autophagic cell death pathways via regulation of miRNA and DRAM in human osteosarcoma cells.
Molecular Cancer Therapeutics 01/2013; · 5.23 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: We report a case of gastric cancer that presented as transverse myelopathy due to spinal bone metastasis. A 45-year- old man with advanced gastric cancer underwent distal gastrectomy and lymph node dissection for curative intent. However, pathological examination of the specimen revealed positive cytological findings in the peritoneal lavage fluid in addition to serosal invasion and lymph node metastasis(pT4aN3bCY1, stage IV). Three months after the operation, during the second course of chemotherapy with S-1, he began to complain of back pain, and positron emission tomography-computed tomography revealed spinal bone metastasis. Despite immediate radiotherapy for the bone metastasis, he soon suffered from paraplegia in the lower extremities followed by disturbances of bladder and bowel function. We created a sigmoid colostomy, which enabled self-care for defecation, and resumed radiotherapy and chemotherapy. Bone metastasis of gastric cancer is rare but the prognosis is very poor. Because of a rapidly deteriorating clinical course, early diagnosis and multidisciplinary approaches are important for gastric cancer patients with spinal metastasis.
Gan to kagaku ryoho. Cancer & chemotherapy 11/2012; 39(12):2357-9.
-
[show abstract]
[hide abstract]
ABSTRACT: A 79-year-old man who had previously undergone partial resection of the remnant stomach and Roux-en-Y reconstruction was diagnosed as having peritoneal recurrence near the ligament of Treitz. In the course of chemotherapy for recurrent gastric cancer, he complained of colic pain. CT examination revealed a marked dilation of the duodenum suggesting the presence of a distal duodenal stricture resulting from the known recurrent tumor. To palliate this intestinal obstruction, we successfully placed an expandable metal stent(EMS) using a double-balloon enteroscope(DBE), which achieved immediate relief of the obstruction and enabled the resumption of oral intake and chemotherapy. While the endoscopic placement of an EMS is available for malignant gastro-intestinal obstruction, it is considerably more difficult to approach the duodenum with Roux-en-Y anastomosis. A DBE has made it possible to place an EMS deep in the small intestine. In the present case, this minimally invasive procedure avoided the need for surgery and greatly contributed to palliation. Thus, EMS placement using a DBE is a possible palliative treatment for malignant small bowel obstruction.
Gan to kagaku ryoho. Cancer & chemotherapy 11/2012; 39(12):2372-4.
-
Hiroshi Tazawa,
Shuya Yano,
Ryosuke Yoshida,
Yasumoto Yamasaki,
Tsuyoshi Sasaki,
Yuuri Hashimoto,
Shinji Kuroda,
Masaaki Ouchi,
Teppei Onishi, Futoshi Uno,
Shunsuke Kagawa,
Yasuo Urata,
Toshiyoshi Fujiwara
[show abstract]
[hide abstract]
ABSTRACT: Autophagy is known to have a cytoprotective role under various cellular stresses; however, it also results in robust cell death as an important safeguard mechanism that protects the organism against invading pathogens and unwanted cancer cells. Autophagy is regulated by cell signalling including microRNA (miRNA), a post-transcriptional regulator of gene expression. Here, we show that genetically engineered telomerase-specific oncolytic adenovirus induced miR-7 expression, which is significantly associated with its cytopathic activity in human cancer cells. Virus-mediated miR-7 upregulation depended on enhanced expression of the E2F1 protein. Ectopic expression of miR-7 suppressed cell viability and induced autophagy by inhibiting epidermal growth factor receptor (EGFR) expression. Our results suggest that oncolytic adenovirus induces autophagic cell death through an E2F1-miR-7-EGFR pathway in human cancer cells, providing a novel insight into the molecular mechanism of an anticancer virotherapy.
International Journal of Cancer 04/2012; 131(12):2939-50. · 5.44 Impact Factor
-
Ryosuke Yoshida,
Hiroshi Tazawa,
Yuuri Hashimoto,
Shuya Yano,
Teppei Onishi,
Tsuyoshi Sasaki,
Yasuhiro Shirakawa,
Hiroyuki Kishimoto, Futoshi Uno,
Masahiko Nishizaki,
Shunsuke Kagawa,
Toshiyoshi Fujiwara
[show abstract]
[hide abstract]
ABSTRACT: Trastuzumab, a humanized antibody targeting HER2, exhibits remarkable therapeutic efficacy against HER2-positive breast and gastric cancers; however, acquired resistance presents a formidable obstacle to long-term tumor responses in the majority of patients. Here, we show the mechanism of resistance to trastuzumab in HER2-positive human cancer cells and explore the molecular sensitization by exogenous expression of HER2-extracellular domain (ECD) in HER2-negative or trastuzumab-resistant human cancer cells. We found that long-term exposure to trastuzumab induced resistance in HER2-positive cancer cells; HER2 expression was downregulated, and antibody-dependent cellular cytotoxicity (ADCC) activity was impaired. We next examined the hypothesis that trastuzumab-resistant cells could be re-sensitized by the transfer of non-functional HER2-ECD. Exogenous HER2-ECD expression induced by the stable transfection of a plasmid vector or infection with a replication-deficient adenovirus vector had no apparent effect on the signaling pathway, but strongly enhanced ADCC activity in low HER2-expressing or trastuzumab-resistant human cancer cells. Our data indicate that restoration of HER2-ECD expression sensitizes HER2-negative or HER2-downregulated human cancer cells to trastuzumab-mediated ADCC, an outcome that has important implications for the treatment of human cancers.
Cancer Immunology and Immunotherapy 03/2012; · 3.70 Impact Factor
-
Yasumoto Yamasaki,
Hiroshi Tazawa,
Yuuri Hashimoto,
Toru Kojima,
Shinji Kuroda,
Shuya Yano,
Ryosuke Yoshida, Futoshi Uno,
Hiroyuki Mizuguchi,
Akira Ohtsuru,
Yasuo Urata,
Shunsuke Kagawa,
Toshiyoshi Fujiwara
[show abstract]
[hide abstract]
ABSTRACT: Oncolytic viruses engineered to replicate in tumour cells but not in normal cells could be used as tumour-specific vectors carrying the therapeutic genes. We previously developed a telomerase-specific oncolytic adenovirus, OBP-301, that causes cell death in human cancer cells with telomerase activities. Here, we further modified OBP-301 to express the wild-type p53 tumour suppressor gene (OBP-702), and investigated whether OBP-702 induces stronger antitumour activity than OBP-301. The antitumour effect of OBP-702 was compared to that of OBP-301 on OBP-301-sensitive (H358 and H460) and OBP-301-resistant (T.Tn and HSC4) human cancer cells. OBP-702 suppressed the viability of both OBP-301-sensitive and OBP-301-resistant cancer cells more efficiently than OBP-301. OBP-702 caused increased apoptosis compared to OBP-301 or a replication-deficient adenovirus expressing the p53 gene (Ad-p53) in H358 and T.Tn cells. Adenovirus E1A-mediated p21 and MDM2 downregulation was involved in the apoptosis caused by OBP-702. Moreover, OBP-702 significantly suppressed tumour growth in subcutaneous tumour xenograft models compared to monotherapy with OBP-301 or Ad-p53. Our data demonstrated that OBP-702 infection expressed adenovirus E1A and then inhibited p21 and MDM2 expression, which in turn efficiently induced apoptotic cell death. This novel apoptotic mechanism suggests that the p53-expressing OBP-702 is a promising antitumour reagent for human cancer and could improve the clinical outcome.
European journal of cancer (Oxford, England: 1990) 01/2012; 48(14):2282-91. · 4.12 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: Hypoxia is a microenvironmental factor that contributes to the invasion, progression and metastasis of tumor cells. Hypoxic tumor cells often show more resistance to conventional chemoradiotherapy than normoxic tumor cells, suggesting the requirement of novel antitumor therapies to efficiently eliminate the hypoxic tumor cells. We previously generated a tumor-specific replication-competent oncolytic adenovirus (OBP-301: Telomelysin), in which the human telomerase reverse transcriptase (hTERT) promoter drives viral E1 expression. Since the promoter activity of the hTERT gene has been shown to be upregulated by hypoxia, we hypothesized that, under hypoxic conditions, the antitumor effect of OBP-301 with the hTERT promoter would be more efficient than that of the wild-type adenovirus 5 (Ad5). In this study, we investigated the antitumor effects of OBP-301 and Ad5 against human cancer cells under a normoxic (20% oxygen) or a hypoxic (1% oxygen) condition. Hypoxic condition induced nuclear accumulation of the hypoxia-inducible factor-1α and upregulation of hTERT promoter activity in human cancer cells. The cytopathic activity of OBP-301 was significantly higher than that of Ad5 under hypoxic condition. Consistent with their cytopathic activity, the replication of OBP-301 was significantly higher than that of Ad5 under the hypoxic condition. OBP-301-mediated E1A was expressed within hypoxic areas of human xenograft tumors in mice. These results suggest that the cytopathic activity of OBP-301 against hypoxic tumor cells is mediated through hypoxia-mediated activation of the hTERT promoter. Regulation of oncolytic adenoviruses by the hTERT promoter is a promising antitumor strategy, not only for induction of tumor-specific oncolysis, but also for efficient elimination of hypoxic tumor cells.
PLoS ONE 01/2012; 7(6):e39292. · 4.09 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: The case is a 75-year-old man, who underwent curative resection for the upper part of stomach cancer (Stage II), and was without a recurrence for two years. The patient had been well until September 2006, when CT examination revealed a 5 cm tumor in the rectovesical pouch. The tumor was histologically diagnosed as a peritoneal metastasis of gastric cancer through CT-guided percutaneous needle biopsy. Chemotherapy with S-1 was initiated, but it resulted in progressive disease. To locally control the growing tumor and avoid rectal stenosis, irradiation was applied for a total of 56 Gy in December 2007 and then followed by chemotherapy with paclitaxel. He obtained a complete response 7-month later, and remains CR for 3 years by receiving paclitaxel chemotherapy. Although chemotherapy is of choice for gastric cancer recurrence, in some cases radiotherapy can play an important role in local control even in peritoneal metastasis.
Gan to kagaku ryoho. Cancer & chemotherapy 11/2011; 38(12):2100-2.
-
Tsuyoshi Sasaki,
Hiroshi Tazawa,
Jo Hasei,
Toshiyuki Kunisada,
Aki Yoshida,
Yuuri Hashimoto,
Shuya Yano,
Ryosuke Yoshida, Futoshi Uno,
Shunsuke Kagawa,
Yuki Morimoto,
Yasuo Urata,
Toshifumi Ozaki,
Toshiyoshi Fujiwara
[show abstract]
[hide abstract]
ABSTRACT: Tumor-specific replication-selective oncolytic virotherapy is a promising antitumor therapy for induction of cell death in tumor cells but not of normal cells. We previously developed an oncolytic adenovirus, OBP-301, that kills human epithelial malignant cells in a telomerase-dependent manner. Recent evidence suggests that nonepithelial malignant cells, which have low telomerase activity, maintain telomere length through alternative lengthening of telomeres (ALT). However, it remains unclear whether OBP-301 is cytopathic for nonepithelial malignant cells. Here, we evaluated the antitumor effect of OBP-301 on human bone and soft tissue sarcoma cells.
The cytopathic activity of OBP-301, coxsackie and adenovirus receptor (CAR) expression, and telomerase activity were examined in 10 bone (OST, U2OS, HOS, HuO9, MNNG/HOS, SaOS-2, NOS-2, NOS-10, NDCS-1, and OUMS-27) and in 4 soft tissue (CCS, NMS-2, SYO-1, and NMFH-1) sarcoma cell lines. OBP-301 antitumor effects were assessed using orthotopic tumor xenograft models. The fiber-modified OBP-301 (termed OBP-405) was used to confirm an antitumor effect on OBP-301-resistant sarcomas.
OBP-301 was cytopathic for 12 sarcoma cell lines but not for the non-CAR-expressing OUMS-27 and NMFH-1 cells. Sensitivity to OBP-301 was dependent on CAR expression and not on telomerase activity. ALT-type sarcomas were also sensitive to OBP-301 because of upregulation of human telomerase reverse transcriptase (hTERT) mRNA following virus infection. Intratumoral injection of OBP-301 significantly suppressed the growth of OST and SYO-1 tumors. Furthermore, fiber-modified OBP-405 showed antitumor effects on OBP-301-resistant OUMS-27 and NMFH-1 cells.
A telomerase-specific oncolytic adenovirus is a promising antitumor reagent for the treatment of bone and soft tissue sarcomas.
Clinical Cancer Research 02/2011; 17(7):1828-38. · 7.74 Impact Factor
-
Shinji Kuroda,
Toshiya Fujiwara,
Yasuhiro Shirakawa,
Yasumoto Yamasaki,
Shuya Yano, Futoshi Uno,
Hiroshi Tazawa,
Yuuri Hashimoto,
Yuichi Watanabe,
Kazuhiro Noma,
Yasuo Urata,
Shunsuke Kagawa,
Toshiyoshi Fujiwara
[show abstract]
[hide abstract]
ABSTRACT: The inability to repair DNA double-strand breaks (DSB) leads to radiosensitization, such that ionizing radiation combined with molecular inhibition of cellular DSB processing may greatly affect treatment of human cancer. As a variety of viral products interact with the DNA repair machinery, oncolytic virotherapy may improve the therapeutic window of conventional radiotherapy. Here, we describe the mechanistic basis for synergy of irradiation and OBP-301 (Telomelysin), an attenuated type-5 adenovirus with oncolytic potency that contains the human telomerase reverse transcriptase promoter to regulate viral replication. OBP-301 infection led to E1B55kDa viral protein expression that degraded the complex formed by Mre11, Rad50, and NBS1, which senses DSBs. Subsequently, the phosphorylation of cellular ataxia-telangiectasia mutated protein was inhibited, disrupting the signaling pathway controlling DNA repair. Thus, tumor cells infected with OBP-301 could be rendered sensitive to ionizing radiation. Moreover, by using noninvasive whole-body imaging, we showed that intratumoral injection of OBP-301 followed by regional irradiation induces a substantial antitumor effect, resulting from tumor cell-specific radiosensitization, in an orthotopic human esophageal cancer xenograft model. These results illustrate the potential of combining oncolytic virotherapy and ionizing radiation as a promising strategy in the management of human cancer.
Cancer Research 11/2010; 70(22):9339-48. · 7.86 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: Although splenectomy is often performed along with en bloc node dissection in gastric cancer surgery, portal/splenic vein thrombosis (PSVT) has been rarely reported. We recently encountered a case of PSVT after a splenectomy was performed during gastric cancer surgery. A 53-year-old woman underwent total gastrectomy, splenectomy, and en bloc regional lymph node dissection for gastric cancer. An uneventful postoperative course ended with abrupt development of a fever and general fatigue. Laboratory tests showed elevated levels of liver transaminases and fibrinogen degenerative products. Contrast-enhanced computed tomography revealed splenic vein thrombosis and partial liver infarction. Immediate anticoagulant treatment resulted in clinical improvement and partial thrombolysis in 2 months. PSVT after splenectomy in haematological disorders has been recognized as a possibly lethal complication. However, it has been underappreciated in cases of splenectomy for gastric cancer. The present case demonstrates the importance of considering PSVT as a possible complication of splenectomy in gastric cancer surgery.
Asian Journal of Surgery 10/2010; 33(4):208-11. · 0.57 Impact Factor
-
Toru Kojima,
Yuichi Watanabe,
Yuuri Hashimoto,
Shinji Kuroda,
Yasumoto Yamasaki,
Shuya Yano,
Masaaki Ouchi,
Hiroshi Tazawa, Futoshi Uno,
Shunsuke Kagawa,
Satoru Kyo,
Hiroyuki Mizuguchi,
Yasuo Urata,
Noriaki Tanaka,
Toshiyoshi Fujiwara
[show abstract]
[hide abstract]
ABSTRACT: The aim of this study was to develop a less invasive way of targeting lymph node metastasis for the treatment of human gastrointestinal cancer. Lymphatic invasion is a major route for cancer cell dissemination, and adequate treatment of locoregional lymph nodes is required for curative treatment in patients with malignancies.
Human telomerase reverse transcription (hTERT) is the catalytic subunit of telomerase, which is highly active in cancer cells but quiescent in most normal somatic cells. OBP-301 (Telomelysin) is an attenuated adenovirus with oncolytic potency that contains the hTERT promoter element to regulate viral replication. We examined whether OBP-301 injected into the primary tumor might be useful for purging micrometastasis from regional lymph nodes in an orthotopic colorectal cancer model.
OBP-301 was intratumorally injected into HT29 tumors orthotopically implanted into the rectum in BALB/c nu/nu mice. By using a highly sensitive quantitative PCR analysis that targets the human-specific Alu sequence, we showed that OBP-301 caused viral spread into the regional lymphatic area and selectively replicated in neoplastic lesions, resulting in tumor-cell-specific death in metastatic lymph nodes. Moreover, although the surgical removal of primary tumors increased the tendency of lymph node metastasis, preoperative intratumoral injection of virus significantly reduced lymph node metastasis.
Our results indicate that intratumoral injection of OBP-301 mediates effective in vivo purging of metastatic tumor cells from regional lymph nodes, which may help optimize treatment of human cancer, especially gastrointestinal malignancies.
Annals of surgery 06/2010; 251(6):1079-86. · 7.90 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: Multimodal approaches combining drugs that differentially function is the most popular regimen for treating human cancer. Understanding the molecular mechanisms underlying the synergistic, potentiative, and antagonistic effects of drug combinations could facilitate the discovery of novel efficacious combinations. We previously showed that telomerase-specific replication-competent adenovirus (Telomelysin, OBP-301), in which the human telomerase reverse transcriptase promoter controls the adenoviral E1 gene expression, induces a selective antitumor effect in human cancer cells. Here, using E1-deleted replication-deficient adenovirus expressing the p53 tumor suppressor gene (Advexin, Ad-p53) and OBP-301, we investigate how these adenoviruses that kill tumor cells with different mechanisms could work in combination on human cancer. We found that E1-deficient Ad-p53 could kill cancer cells more efficiently in the presence of OBP-301 than Ad-p53 alone or OBP-301 alone, because Ad-p53 could become replication-competent by being supplied adenoviral E1 from coinfected OBP-301 in trans. Ad-p53 plus OBP-301 induced high levels of p53 protein expression without p21 induction, resulting in apoptotic cell death documented by active caspase-3 expression with a cytometric bead array and an increased subdiploid apoptotic fraction of the cell cycle. For in vivo evaluation, nude mice xenografted with human lung tumors received intratumoral injection of OBP-301 and/or Ad-p53. Analysis of the growth of implanted tumors showed an enhanced antitumor effect in combination therapy. Our data show that Ad-p53 in combination with OBP-301 induces not only oncolytic but also apoptotic cancer cell death and enhances antitumor activity in vitro and in vivo, providing potential merits as a multimodal treatment for human cancer.
Molecular Cancer Therapeutics 06/2010; 9(6):1884-93. · 5.23 Impact Factor
-
Toru Kojima,
Yuuri Hashimoto,
Yuichi Watanabe,
Shunsuke Kagawa, Futoshi Uno,
Shinji Kuroda,
Hiroshi Tazawa,
Satoru Kyo,
Hiroyuki Mizuguchi,
Yasuo Urata,
Noriaki Tanaka,
Toshiyoshi Fujiwara
[show abstract]
[hide abstract]
ABSTRACT: The presence of circulating tumor cells (CTCs) in the peripheral blood is associated with short survival, making the detection of CTCs clinically useful as a prognostic factor of disease outcome and/or a surrogate marker of treatment response. Recent technical advances in immunocytometric analysis and quantitative real-time PCR have made it possible to detect a few CTCs in the blood; however, there is no sensitive assay to specifically detect viable CTCs. Here, we report what we believe to be a new approach to visually detect live human CTCs among millions of peripheral blood leukocytes, using a telomerase-specific replication-selective adenovirus expressing GFP. First, we constructed a GFP-expressing attenuated adenovirus, in which the telomerase promoter regulates viral replication (OBP-401; TelomeScan). We then used OBP-401 to establish a simple ex vivo method that was able to detect viable human CTCs in the peripheral blood. The detection method involved a 3-step procedure, including the lysis of rbc, the subsequent addition of OBP-401 to the cell pellets, and an automated scan using fluorescence microscopy. OBP-401 infection increased the signal-to-background ratio as a tumor-specific probe, because the fluorescent signal was amplified only in viable, infected human tumor cells, by viral replication. This GFP-expressing virus-based method is remarkably simple and allows precise enumeration of CTCs.
The Journal of clinical investigation 10/2009; 119(10):3172-81. · 15.39 Impact Factor
-
Yoshihiro Ikeda,
Toru Kojima,
Shinji Kuroda,
Yoshikatsu Endo,
Ryo Sakai,
Masayoshi Hioki,
Hiroyuki Kishimoto, Futoshi Uno,
Shunsuke Kagawa,
Yuichi Watanabe,
Yuuri Hashimoto,
Yasuo Urata,
Noriaki Tanaka,
Toshiyoshi Fujiwara
[show abstract]
[hide abstract]
ABSTRACT: Soluble factors in the tumor microenvironment may influence the process of angiogenesis; a process essential for the growth and progression of malignant tumors. In this study, we describe a novel antiangiogenic effect of conditional replication-selective adenovirus through the stimulation of host immune reaction. An attenuated adenovirus (OBP-301, Telomelysin), in which the human telomerase reverse transcriptase promoter element drives expression of E1 genes, could replicate in and cause selective lysis of cancer cells. Mixed lymphocyte-tumor cell culture demonstrated that OBP-301-infected cancer cells stimulated PBMC to produce IFN-gamma into the supernatants. When the supernatants were subjected to the assay of in vitro angiogenesis, the tube formation of HUVECs was inhibited more efficiently than recombinant IFN-gamma. Moreover, in vivo angiogenic assay using a membrane-diffusion chamber system s.c. transplanted in nu/nu mice showed that tumor cell-induced neovascularization was markedly reduced when the chambers contained the mixed lymphocyte-tumor cell culture supernatants. The growth of s.c. murine colon tumors in syngenic mice was significantly inhibited due to the reduced vascularity by intratumoral injection of OBP-301. The antitumor as well as antiangiogenic effects, however, were less apparent in SCID mice due to the lack of host immune responses. Our data suggest that OBP-301 seems to have antiangiogenic properties through the stimulation of host immune cells to produce endogenous antiangiogenic factors such as IFN-gamma.
The Journal of Immunology 03/2009; 182(3):1763-9. · 5.79 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: Cancer gene therapy and oncolytic virotherapy have been studied extensively. However, their clinical application is hampered by their weak anticancer activity. We previously constructed a replicating adenovirus (OBP-301, Telomelysin), in which the human telomerase reverse transcriptase (hTERT) promoter drives expression of the adenoviral E1 genes, and causes selective lysis of human cancer cells. We hypothesized that combination adenoviral therapy containing OBP-301 and a nonreplicating adenovirus expressing the proapoptotic Bax gene could overcome the weakness and augment the anticancer efficacy of each modality. Combination treatment resulted in marked Bax protein expression and enhanced efficacy in in vitro cell viability assay, when compared with either single treatment. However, combination treatment was not as effective in suppressing both subcutaneous and pleural disseminated tumors compared with OBP-301 treatment alone. Further investigation revealed that combination treatment resulted in suppressed E1A protein expression associated with reduced viral replication. Our results suggest that Bax gene therapy in combination with oncolytic adenovirotherapy potentially augments their antitumor activity, but further improvements may be required to maximize the combinatorial effect in vivo, for the Bax gene expression to avoid interference with production of the oncolytic virus.
International Journal of Cancer 07/2008; 122(11):2628-33. · 5.44 Impact Factor
-
Yanan Yang,
Marie Wislez,
Nobukazu Fujimoto,
Ludmila Prudkin,
Julie G Izzo, Futoshi Uno,
Lin Ji,
Amy E Hanna,
Robert R Langley,
Diane Liu,
Faye M Johnson,
Ignacio Wistuba,
Jonathan M Kurie
[show abstract]
[hide abstract]
ABSTRACT: The c-Met receptor tyrosine kinase has been implicated in cellular transformation induced by mutant Ras, a commonly activated proto-oncogene in non-small cell lung cancer (NSCLC). However, the role of c-Met has not been defined in K-ras-mutant NSCLC, a disease for which no effective targeted therapeutic options currently exist. To acquire a greater understanding of its role, we used genetic and pharmacologic approaches to inhibit c-Met in mice and cultured cells. In Kras(LA1) mice, which develop premalignant lung lesions that progress to multifocal lung adenocarcinomas owing to somatic mutations in K-ras, c-Met was expressed in multiple cell types within premalignant lung lesions, and high concentrations of HGF were detected in bronchoalveolar lavage samples. Short-term treatment with PHA-665752, a c-Met inhibitor, decreased the numbers of premalignant lung lesions and induced apoptosis in tumor cells and vascular endothelial cells within lesions. In cell culture, PHA-665752 induced apoptosis of a lung adenocarcinoma cell line derived from Kras(LA1) mice (LKR-13) and a murine lung endothelial cell line (MEC). c-Met depletion by siRNA transfection induced apoptosis of MECs but not LKR-13 cells. Collectively, these findings suggest that apoptosis was an on-target effect of PHA-665752 in MECs but not in LKR-13 cells. We conclude that PHA-665752 inhibited lung tumorigenesis in Kras(LA1) mice and may provide a novel therapeutic approach to the prevention of K-ras-mutant NSCLC.
Molecular Cancer Therapeutics 05/2008; 7(4):952-60. · 5.23 Impact Factor
-
Yuuri Hashimoto,
Yuichi Watanabe,
Yoshiko Shirakiya, Futoshi Uno,
Shunsuke Kagawa,
Hitoshi Kawamura,
Katsuyuki Nagai,
Noriaki Tanaka,
Horomi Kumon,
Yasuo Urata,
Toshiyoshi Fujiwara
[show abstract]
[hide abstract]
ABSTRACT: The use of replication-selective tumor-specific viruses represents a novel approach for the treatment of neoplastic disease. We constructed an attenuated adenovirus, telomerase-specific replication-selective adenovirus (TRAD), in which the human telomerase reverse transcriptase promoter element drives the expression of the E1A and E1B genes linked with an internal ribosome entry site (IRES). Forty-eight hours after TRAD infection at a multiplicity of infection of 1.0, the cell viability of H1299 human lung cancer cells was consistently less than 50% and therefore this procedure could be used as a potency assay to assess the biological activity of TRAD. We also established a quantitative real-time polymerase chain reaction (PCR) analysis with consensus primers for either the adenovirus E1A or IRES sequence. The linear ranges of quantitation with E1A and IRES primers were 10(3)-10(8) and 10(2)-10(8) plaque-forming units/mL in the plasma, respectively. The PCR analysis demonstrated that the levels of E1A in normal tissues were more than 10(3) lower than in the tumors of A549 human lung tumor xenografts in nu/nmicro mice at 28 days after intratumoral injection. Our results suggest that the cell-killing assay against H1299 cells and real-time PCR can be used to assess the biological activity and biodistribution of TRAD in clinical trials.
Cancer Science 03/2008; 99(2):385-90. · 3.33 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: A 47-year-old man with no symptoms was admitted to our hospital for the treatment of NSCLC, which was incidentally detected by an X-ray examination at the mass screening. Computed tomography (CT) of the chest and FDG-PET revealed a 3.6 cm tumor in the right upper lobe with multiple lymphadenopathy in the right mediastinum. Based on these clinical findings, we classified this case as a T2N2M0, stage IIIA NSCLC. The patient consented to and received 2 courses of systemic chemotherapy consisting of cisplatin (CDDP 40 mg/m(2); day 1, 8) and docetaxel (DOC 40 mg/m(2); day 1, 8) combined with concurrent radiation (2 Gy/day; total 46 Gy) with no severe adverse events. His tumors responded well to the treatment, and restaging chest CT showed marked regression of mediastinal lymphadenopathy, and partial response to the lung tumor. Then, acurative surgical resection was performed. Finally, the case was diagnosed as a T1N1M0, stage IIA NSCLC pathologically. Our chemotherapy regimen consisting of CDDP and DOC combined with concurrent radiation might be as potent as neo-adjuvant therapy for clinical stage III NSCLC.
Gan to kagaku ryoho. Cancer & chemotherapy 10/2007; 34(9):1493-5.
