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         headline:Pre-clinical studies of Notch signaling inhibitor RO4929097 in inflammatory breast cancer cells
         description:Basal breast cancer, common among patients presenting with inflammatory breast cancer (IBC), has been shown to be resistant to radiation and enriched in cancer stem cells. The Notch pathway plays an important role in self-renewal of breast cancer stem cells and contributes to inflammatory signaling which promotes the breast cancer stem cell phenotype. Herein, we inhibited Notch signaling using a gamma secretase inhibitor, RO4929097, in an in vitro model that enriches for cancer initiating cells (3D clonogenic assay) and conventional 2D clonogenic assay to compare the effect on radiosensitization of the SUM149 and SUM190 IBC cell lines. RO4929097 downregulated the Notch target genes Hes1, Hey1, and HeyL, and showed a significant reduction in anchorage independent growth in SUM190 and SUM149. However, the putative self-renewal assay mammosphere formation efficiency was increased with the drug. To assess radiosensitization of putative cancer stem cells, cells were exposed to increasing doses of radiation with or without 1 μM RO4929097 in their standard (2D) and self-renewal enriching (3D) culture conditions. In the conventional 2D clonogenic assay, RO4929097 significantly sensitized SUM190 cells to ionizing radiation and has a modest radiosensitization effect in SUM149 cells. In the 3D clonogenic assays, however, a radioprotective effect was seen in both SUM149 and SUM190 cells at higher doses. Both cell lines express IL-6 and IL-8 cytokines known to mediate the efficacy of Notch inhibition and to promote self-renewal of stem cells. We further showed that RO429097 inhibits normal T-cell synthesis of some inflammatory cytokines, including TNF-α, a potential mediator of IL-6 and IL-8 production in the microenvironment. These data suggest that additional targeting agents may be required to selectively target IBC stem cells through Notch inhibition, and that evaluation of microenvironmental influences may shed further light on the potential effects of this inhibitor.
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      headline:Pre-clinical studies of Notch signaling inhibitor RO4929097 in inflammatory breast cancer cells
      description:Basal breast cancer, common among patients presenting with inflammatory breast cancer (IBC), has been shown to be resistant to radiation and enriched in cancer stem cells. The Notch pathway plays an important role in self-renewal of breast cancer stem cells and contributes to inflammatory signaling which promotes the breast cancer stem cell phenotype. Herein, we inhibited Notch signaling using a gamma secretase inhibitor, RO4929097, in an in vitro model that enriches for cancer initiating cells (3D clonogenic assay) and conventional 2D clonogenic assay to compare the effect on radiosensitization of the SUM149 and SUM190 IBC cell lines. RO4929097 downregulated the Notch target genes Hes1, Hey1, and HeyL, and showed a significant reduction in anchorage independent growth in SUM190 and SUM149. However, the putative self-renewal assay mammosphere formation efficiency was increased with the drug. To assess radiosensitization of putative cancer stem cells, cells were exposed to increasing doses of radiation with or without 1 μM RO4929097 in their standard (2D) and self-renewal enriching (3D) culture conditions. In the conventional 2D clonogenic assay, RO4929097 significantly sensitized SUM190 cells to ionizing radiation and has a modest radiosensitization effect in SUM149 cells. In the 3D clonogenic assays, however, a radioprotective effect was seen in both SUM149 and SUM190 cells at higher doses. Both cell lines express IL-6 and IL-8 cytokines known to mediate the efficacy of Notch inhibition and to promote self-renewal of stem cells. We further showed that RO429097 inhibits normal T-cell synthesis of some inflammatory cytokines, including TNF-α, a potential mediator of IL-6 and IL-8 production in the microenvironment. These data suggest that additional targeting agents may be required to selectively target IBC stem cells through Notch inhibition, and that evaluation of microenvironmental influences may shed further light on the potential effects of this inhibitor.
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            address:
               name:Department of Radiation Oncology, Unit 1202, The University of Texas MD Anderson Cancer Center, Houston, USA
               type:PostalAddress
            type:Organization
            name:The University of Texas MD Anderson Cancer Center
            address:
               name:Morgan Welch Inflammatory Breast Cancer Research Center and Clinic, The University of Texas MD Anderson Cancer Center, Houston, USA
               type:PostalAddress
            type:Organization
      name:Evan N. Cohen
      affiliation:
            name:The University of Texas MD Anderson Cancer Center
            address:
               name:Department of Hematopathology, The University of Texas MD Anderson Cancer Center, Houston, USA
               type:PostalAddress
            type:Organization
            name:The University of Texas MD Anderson Cancer Center
            address:
               name:Morgan Welch Inflammatory Breast Cancer Research Center and Clinic, The University of Texas MD Anderson Cancer Center, Houston, USA
               type:PostalAddress
            type:Organization
      name:Kimberly Boley
      affiliation:
            name:The University of Texas MD Anderson Cancer Center
            address:
               name:Department of Experimental Therapeutics, The University of Texas MD Anderson Cancer Center, Houston, USA
               type:PostalAddress
            type:Organization
            name:The University of Texas MD Anderson Cancer Center
            address:
               name:Morgan Welch Inflammatory Breast Cancer Research Center and Clinic, The University of Texas MD Anderson Cancer Center, Houston, USA
               type:PostalAddress
            type:Organization
      name:Erik M. Freiter
      affiliation:
            name:The University of Texas MD Anderson Cancer Center
            address:
               name:Department of Experimental Therapeutics, The University of Texas MD Anderson Cancer Center, Houston, USA
               type:PostalAddress
            type:Organization
            name:The University of Texas MD Anderson Cancer Center
            address:
               name:Morgan Welch Inflammatory Breast Cancer Research Center and Clinic, The University of Texas MD Anderson Cancer Center, Houston, USA
               type:PostalAddress
            type:Organization
      name:Li Li
      affiliation:
            name:The University of Texas MD Anderson Cancer Center
            address:
               name:Department of Radiation Oncology, Unit 1202, The University of Texas MD Anderson Cancer Center, Houston, USA
               type:PostalAddress
            type:Organization
            name:The University of Texas MD Anderson Cancer Center
            address:
               name:Morgan Welch Inflammatory Breast Cancer Research Center and Clinic, The University of Texas MD Anderson Cancer Center, Houston, USA
               type:PostalAddress
            type:Organization
      name:Fredika M. Robertson
      affiliation:
            name:The University of Texas MD Anderson Cancer Center
            address:
               name:Department of Experimental Therapeutics, The University of Texas MD Anderson Cancer Center, Houston, USA
               type:PostalAddress
            type:Organization
            name:The University of Texas MD Anderson Cancer Center
            address:
               name:Morgan Welch Inflammatory Breast Cancer Research Center and Clinic, The University of Texas MD Anderson Cancer Center, Houston, USA
               type:PostalAddress
            type:Organization
      name:James M. Reuben
      affiliation:
            name:The University of Texas MD Anderson Cancer Center
            address:
               name:Department of Hematopathology, The University of Texas MD Anderson Cancer Center, Houston, USA
               type:PostalAddress
            type:Organization
            name:The University of Texas MD Anderson Cancer Center
            address:
               name:Morgan Welch Inflammatory Breast Cancer Research Center and Clinic, The University of Texas MD Anderson Cancer Center, Houston, USA
               type:PostalAddress
            type:Organization
      name:Massimo Cristofanilli
      affiliation:
            name:Fox Chase Cancer Center
            address:
               name:Fox Chase Cancer Center, Philadelphia, USA
               type:PostalAddress
            type:Organization
            name:The University of Texas MD Anderson Cancer Center
            address:
               name:Morgan Welch Inflammatory Breast Cancer Research Center and Clinic, The University of Texas MD Anderson Cancer Center, Houston, USA
               type:PostalAddress
            type:Organization
      name:Thomas A. Buchholz
      affiliation:
            name:The University of Texas MD Anderson Cancer Center
            address:
               name:Department of Radiation Oncology, Unit 1202, The University of Texas MD Anderson Cancer Center, Houston, USA
               type:PostalAddress
            type:Organization
            name:The University of Texas MD Anderson Cancer Center
            address:
               name:Morgan Welch Inflammatory Breast Cancer Research Center and Clinic, The University of Texas MD Anderson Cancer Center, Houston, USA
               type:PostalAddress
            type:Organization
      name:Wendy A. Woodward
      affiliation:
            name:The University of Texas MD Anderson Cancer Center
            address:
               name:Department of Radiation Oncology, Unit 1202, The University of Texas MD Anderson Cancer Center, Houston, USA
               type:PostalAddress
            type:Organization
            name:The University of Texas MD Anderson Cancer Center
            address:
               name:Morgan Welch Inflammatory Breast Cancer Research Center and Clinic, The University of Texas MD Anderson Cancer Center, Houston, USA
               type:PostalAddress
            type:Organization
      email:[email protected]
PostalAddress:
      name:Department of Radiation Oncology, Unit 1202, The University of Texas MD Anderson Cancer Center, Houston, USA
      name:Morgan Welch Inflammatory Breast Cancer Research Center and Clinic, The University of Texas MD Anderson Cancer Center, Houston, USA
      name:Department of Hematopathology, The University of Texas MD Anderson Cancer Center, Houston, USA
      name:Morgan Welch Inflammatory Breast Cancer Research Center and Clinic, The University of Texas MD Anderson Cancer Center, Houston, USA
      name:Department of Experimental Therapeutics, The University of Texas MD Anderson Cancer Center, Houston, USA
      name:Morgan Welch Inflammatory Breast Cancer Research Center and Clinic, The University of Texas MD Anderson Cancer Center, Houston, USA
      name:Department of Experimental Therapeutics, The University of Texas MD Anderson Cancer Center, Houston, USA
      name:Morgan Welch Inflammatory Breast Cancer Research Center and Clinic, The University of Texas MD Anderson Cancer Center, Houston, USA
      name:Department of Radiation Oncology, Unit 1202, The University of Texas MD Anderson Cancer Center, Houston, USA
      name:Morgan Welch Inflammatory Breast Cancer Research Center and Clinic, The University of Texas MD Anderson Cancer Center, Houston, USA
      name:Department of Experimental Therapeutics, The University of Texas MD Anderson Cancer Center, Houston, USA
      name:Morgan Welch Inflammatory Breast Cancer Research Center and Clinic, The University of Texas MD Anderson Cancer Center, Houston, USA
      name:Department of Hematopathology, The University of Texas MD Anderson Cancer Center, Houston, USA
      name:Morgan Welch Inflammatory Breast Cancer Research Center and Clinic, The University of Texas MD Anderson Cancer Center, Houston, USA
      name:Fox Chase Cancer Center, Philadelphia, USA
      name:Morgan Welch Inflammatory Breast Cancer Research Center and Clinic, The University of Texas MD Anderson Cancer Center, Houston, USA
      name:Department of Radiation Oncology, Unit 1202, The University of Texas MD Anderson Cancer Center, Houston, USA
      name:Morgan Welch Inflammatory Breast Cancer Research Center and Clinic, The University of Texas MD Anderson Cancer Center, Houston, USA
      name:Department of Radiation Oncology, Unit 1202, The University of Texas MD Anderson Cancer Center, Houston, USA
      name:Morgan Welch Inflammatory Breast Cancer Research Center and Clinic, The University of Texas MD Anderson Cancer Center, Houston, USA
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