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Dose finding in vivo study of IMD-0354.

Supplemental Figure Legend.

The prolongation of progression-free survival (PFS) in patients with advanced ovarian cancer by antiangiogenic therapy has been shown in several clinical trials. However, although an anti-VEGF antibody (bevacizumab) is the only option currently available, its efficacy is limited and it is not cost effective for use in all patients. Therefore, the development of a novel antiangiogenic drug, especially composed of small-molecule compounds, could be a powerful armament for ovarian cancer treatment. As NF-κB signaling has the potential to regulate VEGF expression, we determined to identify whether VEGF expression is associated with NF-κB activation and to investigate the possibility of a novel IKKβ inhibitor, IMD-0354 (IMMD Inc.), as an antiangiogenic drug. Tissue microarrays from 94 ovarian cancer tissues were constructed and immunohistochemical analyses performed. We revealed that IKK phosphorylation is an independent prognostic factor (PFS: 26.1 vs. 49.8 months, P = 0.011), and is positively correlated with high VEGF expression. In in vitro analyses, IMD-0354 robustly inhibited adhesive and invasive activities of ovarian cancer cells without impairing cell viabilities. IMD-0354 significantly suppressed VEGF production from cancer cells, which led to the inhibition of angiogenesis. In a xenograft model, the treatment of IMD-0354 significantly inhibited peritoneal dissemination with a marked reduction of intratumoral blood vessel formation followed by the inhibition of VEGF expression from cancer cells. IMD-0354 is a stable small-molecule drug and has already been administered safely to humans in other trials. Antiangiogenic therapy targeting IKKβ is a potential future option to treat ovarian cancer. Mol Cancer Ther; 14(4); 909–19. ©2015 AACR.

Characteristics of 94 ovarian cancer patients.

Multivariate analysis for progression free survival of patients.

A new integration technique of a 45-degree micromirror providing a vertical coupling between a free-space wave and a guided wave in a dielectric-glass waveguide for high-density intra-board optical interconnection was described. A planar waveguide consisting of a 4-μm-thickness GeO :SiO guiding core layer and a 2-μm-thickness SiO cladding layer on an SiO substrate was used for characterization of the micromirror. A trench with 8-μm depth and 8-μm width was formed in the waveguide by using a dry etching technique. A photoresist filling the trench was exposed at an angle of 45 degrees in the water to give a 45-degree taper in the trench. Au was evaporated on the taper to give high-reflection micromirror. An excess loss due to the micromirror insertion was estimated to be about 2 dB by comparing insertion losses of waveguides with and without the micromirror.

Purpose: Calponin h1 (CNh1), one of the family of actin-binding proteins, stabilizes the filaments of actin and modulates various cellular biological phenotypes. Recent studies revealed the close correlation between the invasive tumor spread and the reduced expression of CNh1 and α-smooth muscle actin in the surrounding stromal cells. The purpose of this study is to evaluate the efficacy of i.p. CNh1 gene therapy against peritoneal dissemination of ovarian cancer. Experimental Design: We used an adenoviral vector to induce the CNh1 gene into peritoneal cells and ovarian cancer cells as a means of enhancing or inducing the expression of α-smooth muscle actin as well as CNh1. The efficacy of gene transfer was examined by in vitro cell culture and in vivo animal experiments. Results: The formation of longer and thicker actin fibers was observed in each transfected cell line, and the localization of these fibers coincided with that of externally transducted CNh1. With respect to changes in cell behavior, the CNh1-transfected peritoneal cells acquired an ability to resist ovarian cancer-induced shrinkage in cell shape; thus, cancer cell invasion through the monolayer of peritoneal cells was inhibited. In addition, CNh1-transfected ovarian cancer cells showed suppressed anchorage-independent growth and invasiveness, the latter of which accompanied impaired cell motility. The concomitant CNh1 transfection into both peritoneal cells and ovarian cancer cells produced an additive inhibitory effect with respect to cancer cell invasion through the peritoneal cell monolayer. By in vivo experiments designed to treat nude mice that had been i.p. inoculated with ovarian cancer cells, we found that the i.p. injected CNh1 adenovirus successfully blocked cancer-induced morphologic changes in peritoneal cell surface and significantly prolonged the survival time of tumor-bearing mice. Moreover, CNh1 adenovirus could successfully enhance the therapeutic effect of an anticancer drug without increase in side effects. Conclusions: Thus, CNh1 gene therapy against peritoneal dissemination of ovarian cancer is bifunctionally effective (i.e., through inhibitory effects on the infected peritoneal cell layers that suppress cancer invasion and through direct antitumor effects against invasion and growth properties of cancer cells).

We have developed a metabolic profiling scheme based on direct-infusion Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR/MS). The scheme consists of: (1) reproducible data collection under optimized FT-ICR/MS analytical conditions; (2) automatic mass-error correction and multivariate analyses for metabolome characterization using a newly developed metabolomics tool (DMASS software); (3) identification of marker metabolite candidates by searching a species-metabolite relationship database, KNApSAcK; and (4) structural analyses by an MS/MS method. The scheme was applied to metabolic phenotyping of Arabidopsis (Arabidopsis thaliana) seedlings treated with different herbicidal chemical classes for pathway-specific inhibitions. Arabidopsis extracts were directly infused into an electrospray ionization source on an FT-ICR/MS system. Acquired metabolomics data were comprised of mass-to-charge ratio values with ion intensity information subjected to principal component analysis, and metabolic phenotypes from the herbicide treatments were clearly differentiated from those of the herbicide-free treatment. From each herbicide treatment, candidate metabolites representing such metabolic phenotypes were found through the KNApSAcK database search. The database search and MS/MS analyses suggested dose-dependent accumulation patterns of specific metabolites including several flavonoid glycosides. The metabolic phenotyping scheme on the basis of FT-ICR/MS coupled with the DMASS program is discussed as a general tool for high throughput metabolic phenotyping studies.

In order to establish a more faithful model of clinically recurrent ovarian cancer after paclitaxel-based chemotherapy, a paclitaxel-resistant human ovarian cancer cell line was established in vivo, and its biological profiles were compared with the conventional in vitro established drug-resistant cell line.An in vivo paclitaxel-resistant subline (OM1/Tvivo) was established from the parental human ovarian cancer cell line (OVMG1) by repeated paclitaxel administration into tumor-bearing mice. As a control, the in vivo drug-sensitive subline (OM1/Cvivo) was made in the same manner, without paclitaxel. An in vitro paclitaxel-resistant subline (OM1/Tvitro) was established by exposure to stepwise increased concentrations of the drug in a cell culture medium. Chromosomal analysis, evaluation of growth, invasiveness and metastasis, in vivo and in vitro drug sensitivity, and a pharmacokinetic study were performed.Both in vivo sublines confirmed their human origin by G-band chromosomal analysis and showed a similar cell growth rate in cell culture. As for in vivo tumor growth, OM1/Tvivo showed enhanced tumor growth property compared with OM1/Cvivo, while OM1/Tvitro lost tumorigenicity. Both OM1/Tvivo and OM1/Cvivo sublines as well as their parental OVMG1 could not form either invasive or metastatic lesions. Compared with the OM1/Cvivo subline, the OM1/Tvivo tumor showed stable drug-resistance and lower drug distribution after paclitaxel administration into mice, whereas cultured OM1/Tvivo cells lost both completely. On the other hand, an unreasonably higher level of drug-resistance and lower drug concentration was detected in vitro only in OM1/Tvitro cells after exposure to the drug in a culture medium.These results suggest that the in vivo established paclitaxel-resistant cell line, rather than the conventional in vitro established cell line, is a suitable and faithful model for clinically recurrent tumors showing transformed aggressiveness. The in vivo specific drug-resistant mechanism should involve an interaction between the tumor and host stromal tissue rather than only changes in cellular drug sensitivity. The present study is probably the first report of an in vivo established paclitaxel-resistant human ovarian cancer cell line, and the elucidation of such an in vivo drug-resistance mechanism may be clinically important in preventing or overcoming acquired drug-resistant ovarian cancers recurring after paclitaxel-based chemotherapy.

Ovarian cancer remains the most lethal gynecologic cancer and new targeted molecular therapies against this miserable disease continue to be challenging. In this study, we analyzed the expressional patterns of Interleukin-6 (IL-6) and its receptor (IL-6R) expression in ovarian cancer tissues, evaluated the impact of these expressions on clinical outcomes of patients, and found that a high-level of IL-6R expression but not IL-6 expression in cancer cells is an independent prognostic factor. In in vitro analyses using ovarian cell lines, while six (RMUG-S, RMG-1, OVISE, A2780, SKOV3ip1 and OVCAR-3) of seven overexpressed IL-6R compared with a primary normal ovarian surface epithelium, only two (RMG-1, OVISE) of seven cell lines overexpressed IL-6, suggesting that IL-6/IL-6R signaling exerts in a paracrine manner in certain types of ovarian cancer cells. Ovarian cancer ascites were collected from patients, and we found that primary CD11b+CD14+ cells, which were predominantly M2-polarized macrophages, are the major source of IL-6 production in an ovarian cancer microenvironment. When CD11b+CD14+ cells were co-cultured with cancer cells, both the invasion and the proliferation of cancer cells were robustly promoted and these promotions were almost completely inhibited by pretreatment with anti-IL-6R antibody (tocilizumab). The data presented herein suggest a rationale for anti-IL-6/IL-6R therapy to suppress the peritoneal spread of ovarian cancer, and represent evidence of the therapeutic potential of anti-IL-6R therapy for ovarian cancer treatment.

The prolongation of progression-free survival (PFS) in patients with advanced ovarian cancer by antiangiogenic therapy has been shown in several clinical trials. However, although an anti-VEGF antibody (bevacizumab) is the only option currently available, its efficacy is limited and it is not cost effective for use in all patients. Therefore, the development of a novel antiangiogenic drug, especially composed of small-molecule compounds, could be a powerful armament for ovarian cancer treatment. As NF-κB signaling has the potential to regulate VEGF expression, we determined to identify whether VEGF expression is associated with NF-κB activation and to investigate the possibility of a novel IKKβ inhibitor, IMD-0354 (IMMD Inc.), as an antiangiogenic drug. Tissue microarrays from 94 ovarian cancer tissues were constructed and immunohistochemical analyses performed. We revealed that IKK phosphorylation is an independent prognostic factor (PFS: 26.1 vs. 49.8 months, P = 0.011), and is positively correlated with high VEGF expression. In in vitro analyses, IMD-0354 robustly inhibited adhesive and invasive activities of ovarian cancer cells without impairing cell viabilities. IMD-0354 significantly suppressed VEGF production from cancer cells, which led to the inhibition of angiogenesis. In a xenograft model, the treatment of IMD-0354 significantly inhibited peritoneal dissemination with a marked reduction of intratumoral blood vessel formation followed by the inhibition of VEGF expression from cancer cells. IMD-0354 is a stable small-molecule drug and has already been administered safely to humans in other trials. Antiangiogenic therapy targeting IKKβ is a potential future option to treat ovarian cancer. Mol Cancer Ther; 14(4); 909–19. ©2015 AACR.

A surface grating integrated in a channel waveguide on a high-reflection layer is discussed to provide a mirror having a large dependence of a reflection phase variation on wavelength.

Surface mount packaging of a vertical-cavity surface-emitting laser (VCSEL) on an optical board and the input coupling of the wave launched from the VCSEL into a small-number-mode waveguide integrated in the optical board are key issues to construct a future high-density on-board optical interconnect system. A problem in practical applications is too high precision alignment and stability in packaging. Potential solutions would include a combination of a half-VCSEL and an external mirror integrated in the waveguide. A new type of mirror named a cavity-resonator-integrated guided-mode-resonance mirror (CRIGM) is investigated to serve as a laser mirror as wall as an input coupler because it can provide both functions of high reflectivity and simultaneous excitation of the guided mode. A reflection-phase variation of CRIGM takes an important role to construct such a VCSEL. In this paper, a wavelength dependence of the reflection phase is discussed theoretically and demonstrated experimentally.

A cavity-resonator-integrated guided-mode resonance filter (CRIGF) consisting of a grating coupler and a pair of distributed Bragg reflectors on a thin film waveguide has been recently proposed and demonatrated to provide a narrow-band reflection spectrum for an incident wave of a small beam width from the free space. CRIGF is attractive for constructing a very compact laser system. Since a reflection phase of CRIGF is a crucial factor in designing it as a laser mirror, CRIGF with a wavelength dependence of the reflection phase is discussed in this paper. A reflection spectrum was calculated by the finite-difference time-domain (FDTD) method for CRIGF with a resonance wavelength of 848 nm and an aperture size of 10 μm. The reflectance has the maximum of about 60% at the resonance wavelength but decreases to 0% at a slightly different wavelength. Whereas the reflection of the maximum reflectance can be explained by the cancellation of transmission with anti-phase relation of the radiation wave to the direct transmitted wave, the reflection of 0% would indicate in-phase relation. It is predicted that the reflection phase changes between the two wavelengths. A reflection phase variation depending on wavelength was confirmed and estimated to be π/2 by FDTD simulation and optical measurement.

Objective Although brain metastases from gynecologic malignancies are rare, such cases have been gradually increasing in number. The aim of the present study was to evaluate the clinicopathologic features and prognostic factors of brain metastases from gynecologic malignancies. Methods Retrospective analysis of 139 patients with brain metastases from gynecologic malignancies was carried out as a multi-institutional study. The clinicophathological data of the patients were collected from medical records. Results Median survival time of the patients with brain metastases was 12.5months for the ovarian cancer group, 6.2months for the corpus cancer group, and 5.0months for the cervical cancer group; two-year overall survival rates were 19.7%, 6.1%, and 4.8%, respectively. Multivariate analysis revealed ovarian/tubal/peritoneal origin, KPS >70, single brain metastasis, absence of extracranial disease, cranial surgery, cranial radiotherapy, and chemotherapy to be independent favorable prognostic factors associated with overall survival. Conclusion It is considered that aggressive multimodal therapy is warranted in the treatment of brain metastases from gynecologic malignancies in carefully selected patients. The present study may provide a platform for the discussion of management strategies in these rare clinical scenarios.

We report two cases of a complete hydatidiform mole coexistent with a live fetus diagnosed by DNA polymorphism analysis. A 27-year-old woman revealed symptoms of pre-eclampsia and ultrasound showed multicystic tumor and placenta coexistent with a live fetus at 16 weeks' gestation. The placenta with partly hydropic change and the fetus without anomaly were consequently evacuated. Another 30-year-old woman had a multicystic mass attached to a normal placenta with a 20-week live fetus on ultrasound examination. A hysterotomy was carried out because of persistent bleeding due to placenta previa. In both cases, DNA was extracted from the placental tissue and the tumor, as well as from maternal and paternal blood. Genetic analysis demonstrated that the placental tumor consisted of only paternal origin, which is consistent with the diagnosis of complete hydatidiform mole.

A new type of mirror, based on guided-mode resonance, was proposed and discussed to provide a mirror having high reflectance and large wavelength dependence of reflection phase variation. The proposed mirror consists of a surface grating integrated in a channel waveguide on a high-reflection layer. A SiO2-based device was fabricated for 0.85-μm wavelength operation, and reflection phase variation of almost π, with wavelength change of sub-nanometers, was confirmed experimentally.