36 results on '"Bouillez A"'
Search Results
2. Data from MUC1-C Represses the Crumbs Complex Polarity Factor CRB3 and Downregulates the Hippo Pathway
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Donald Kufe, Yozo Suzuki, Masayuki Hiraki, Takahiro Maeda, Hasan Rajabi, Rehan Ahmad, Ashujit Tagde, Audrey Bouillez, and Maroof Alam
- Abstract
Apical–basal polarity and epithelial integrity are maintained in part by the Crumbs (CRB) complex. The C-‐terminal subunit of MUC1 (MUC1-C) is a transmembrane protein that is expressed at the apical border of normal epithelial cells and aberrantly at high levels over the entire surface of their transformed counterparts. However, it is not known whether MUC1-C contributes to this loss of polarity that is characteristic of carcinoma cells. Here it is demonstrated that MUC1-C downregulates expression of the Crumbs complex CRB3 protein in triple-negative breast cancer (TNBC) cells. MUC1-C associates with ZEB1 on the CRB3 promoter and represses CRB3 transcription. Notably, CRB3 activates the core kinase cassette of the Hippo pathway, which includes LATS1 and LATS2. In this context, targeting MUC1-C was associated with increased phosphorylation of LATS1, consistent with activation of the Hippo pathway, which is critical for regulating cell contact, tissue repair, proliferation, and apoptosis. Also shown is that MUC1-C‐-mediated suppression of CRB3 and the Hippo pathway is associated with dephosphorylation and activation of the oncogenic YAP protein. In turn, MUC1-C interacts with YAP, promotes formation of YAP/β-catenin complexes, and induces the WNT target gene MYC. These data support a previously unrecognized pathway in which targeting MUC1-C in TNBC cells (i) induces CRB3 expression, (ii) activates the CRB3-driven Hippo pathway, (iii) inactivates YAP, and thereby (iv) suppresses YAP/β-catenin–mediated induction of MYC expression.Implications: These findings demonstrate a previously unrecognized role for the MUC1-C oncoprotein in the regulation of polarity and the Hippo pathway in breast cancer. Mol Cancer Res; 14(12); 1266–76. ©2016 AACR.
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- 2023
3. Supplemental Figure 1 from MUC1-C Represses the Crumbs Complex Polarity Factor CRB3 and Downregulates the Hippo Pathway
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Donald Kufe, Yozo Suzuki, Masayuki Hiraki, Takahiro Maeda, Hasan Rajabi, Rehan Ahmad, Ashujit Tagde, Audrey Bouillez, and Maroof Alam
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MUC1-C regulates CRB3 expression.
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- 2023
4. Figure S1 and Legend from MUC1-C Induces PD-L1 and Immune Evasion in Triple-Negative Breast Cancer
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Donald Kufe, Kunihiko Hinohara, Tsuyoshi Hata, Masaaki Miyo, Yozo Suzuki, Xiufeng Hu, Audrey Bouillez, Maroof Alam, Ashujit Tagde, Hasan Rajabi, Caining Jin, Masayuki Hiraki, and Takahiro Maeda
- Abstract
This file contains the results of studies with DOX-treated (i) BT-549 cells expressing a tet-CshRNA or tet-MUC1shRNA (S1A-S1D), (ii) MDA-MB-231 cells expressing a tet-CshRNA (S1E), and (iii) SUM-159 cells expressing a tet-MUC1shRNA (S1F).
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- 2023
5. Table S2 from MUC1-C Induces PD-L1 and Immune Evasion in Triple-Negative Breast Cancer
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Donald Kufe, Kunihiko Hinohara, Tsuyoshi Hata, Masaaki Miyo, Yozo Suzuki, Xiufeng Hu, Audrey Bouillez, Maroof Alam, Ashujit Tagde, Hasan Rajabi, Caining Jin, Masayuki Hiraki, and Takahiro Maeda
- Abstract
This file contains ChIP qPCR primer pairs for the GAPDH and PD-L1 promoters.
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- 2023
6. Figure S2 and Legend from MUC1-C Induces PD-L1 and Immune Evasion in Triple-Negative Breast Cancer
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Donald Kufe, Kunihiko Hinohara, Tsuyoshi Hata, Masaaki Miyo, Yozo Suzuki, Xiufeng Hu, Audrey Bouillez, Maroof Alam, Ashujit Tagde, Hasan Rajabi, Caining Jin, Masayuki Hiraki, and Takahiro Maeda
- Abstract
This file contains the results obtained from JQ1 treatment of BT-549 (S2A) and BT-20/MUC1-C (S2B), which demonstrate that JQ1 decreases MYC and PD-L1 expression.
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- 2023
7. Data from MUC1-C Induces PD-L1 and Immune Evasion in Triple-Negative Breast Cancer
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Donald Kufe, Kunihiko Hinohara, Tsuyoshi Hata, Masaaki Miyo, Yozo Suzuki, Xiufeng Hu, Audrey Bouillez, Maroof Alam, Ashujit Tagde, Hasan Rajabi, Caining Jin, Masayuki Hiraki, and Takahiro Maeda
- Abstract
The immune checkpoint ligand PD-L1 and the transmembrane mucin MUC1 are upregulated in triple-negative breast cancer (TNBC), where they contribute to its aggressive pathogenesis. Here, we report that genetic or pharmacological targeting of the oncogenic MUC1 subunit MUC1-C is sufficient to suppress PD-L1 expression in TNBC cells. Mechanistic investigations showed that MUC1-C acted to elevate PD-L1 transcription by recruitment of MYC and NF-κB p65 to the PD-L1 promoter. In an immunocompetent model of TNBC in which Eo771/MUC1-C cells were engrafted into MUC1 transgenic mice, we showed that targeting MUC1-C associated with PD-L1 suppression, increases in tumor-infiltrating CD8+ T cells and tumor cell killing. MUC1 expression in TNBCs also correlated inversely with CD8, CD69, and GZMB, and downregulation of these markers associated with decreased survival. Taken together, our findings show how MUC1 contributes to immune escape in TNBC, and they offer a rationale to target MUC1-C as a novel immunotherapeutic approach for TNBC treatment.Significance: These findings show how upregulation of the transmembrane mucin MUC1 contributes to immune escape in an aggressive form of breast cancer, with potential implications for a novel immunotherapeutic approach. Cancer Res; 78(1); 205–15. ©2017 AACR.
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- 2023
8. Figure S4 and Legend from MUC1-C Induces PD-L1 and Immune Evasion in Triple-Negative Breast Cancer
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Donald Kufe, Kunihiko Hinohara, Tsuyoshi Hata, Masaaki Miyo, Yozo Suzuki, Xiufeng Hu, Audrey Bouillez, Maroof Alam, Ashujit Tagde, Hasan Rajabi, Caining Jin, Masayuki Hiraki, and Takahiro Maeda
- Abstract
This file contains the results of studies with Eo771 cells stably expressing MUC1-C (S4A and S4B), demonstrating that treatment of these cells with JQ1 (S4C), BAY-11-7085 (S4D) or GO-203/NPs (S4E) decreases PD-L1 expression.
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- 2023
9. Figure S3 and Legend from MUC1-C Induces PD-L1 and Immune Evasion in Triple-Negative Breast Cancer
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Donald Kufe, Kunihiko Hinohara, Tsuyoshi Hata, Masaaki Miyo, Yozo Suzuki, Xiufeng Hu, Audrey Bouillez, Maroof Alam, Ashujit Tagde, Hasan Rajabi, Caining Jin, Masayuki Hiraki, and Takahiro Maeda
- Abstract
This file contains the results of studies with DOX-treated BT-549/tet-MUC1shRNA and MDA-MB-231/tet-MUC1shRNA cells, demonstrating that targeting MUC1-C decreases activation of NF-κB p65 (S3A and S3B). The studies also show that targeting NF-κB p65 with a p65shRNA (S3C and S3D) or the BAY-11-7085 inhibitor (S3E and S3F) decreases PD-L1 expression.
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- 2023
10. Figure S5 and Legend from MUC1-C Induces PD-L1 and Immune Evasion in Triple-Negative Breast Cancer
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Donald Kufe, Kunihiko Hinohara, Tsuyoshi Hata, Masaaki Miyo, Yozo Suzuki, Xiufeng Hu, Audrey Bouillez, Maroof Alam, Ashujit Tagde, Hasan Rajabi, Caining Jin, Masayuki Hiraki, and Takahiro Maeda
- Abstract
This file contains results showing that treatment of Eo771/MUC1-C tumors with GO-203/NPs increases CD69 (S5A) and granzyme B (S5B) expression in tumor cells.
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- 2023
11. Figure S6 and Legend from MUC1-C Induces PD-L1 and Immune Evasion in Triple-Negative Breast Cancer
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Donald Kufe, Kunihiko Hinohara, Tsuyoshi Hata, Masaaki Miyo, Yozo Suzuki, Xiufeng Hu, Audrey Bouillez, Maroof Alam, Ashujit Tagde, Hasan Rajabi, Caining Jin, Masayuki Hiraki, and Takahiro Maeda
- Abstract
This file contains results showing that treatment of Eo771/MUC1-C tumors with GO-203/NPs activates tumor infiltrating CD8+ T-cells as evidenced by increases in IFN-γ (S6A), CD107α (S6B) and granzyme B (S6C).
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- 2023
12. Supplementary Tables 1 through 3 from Inhibition of MUC1-C Suppresses MYC Expression and Attenuates Malignant Growth in KRAS Mutant Lung Adenocarcinomas
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Donald Kufe, Kwok-Kin Wong, Ashujit Tagde, Akriti Kharbanda, Maroof Alam, Caining Jin, Sean Pitroda, Hasan Rajabi, and Audrey Bouillez
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qPCR Primers (S1); Promoter ChIP qPCR Primers (S2); Combination Indices (S3).
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- 2023
13. Supplementary Figure 3 from Inhibition of MUC1-C Suppresses MYC Expression and Attenuates Malignant Growth in KRAS Mutant Lung Adenocarcinomas
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Donald Kufe, Kwok-Kin Wong, Ashujit Tagde, Akriti Kharbanda, Maroof Alam, Caining Jin, Sean Pitroda, Hasan Rajabi, and Audrey Bouillez
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Rescue of A549/CRISPR cells with enforced MUC1-C expression.
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- 2023
14. Data from Inhibition of MUC1-C Suppresses MYC Expression and Attenuates Malignant Growth in KRAS Mutant Lung Adenocarcinomas
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Donald Kufe, Kwok-Kin Wong, Ashujit Tagde, Akriti Kharbanda, Maroof Alam, Caining Jin, Sean Pitroda, Hasan Rajabi, and Audrey Bouillez
- Abstract
Dysregulation of MYC expression is a hallmark of cancer, but the development of agents that target MYC has remained challenging. The oncogenic MUC1-C transmembrane protein is, like MYC, aberrantly expressed in diverse human cancers. The present studies demonstrate that MUC1-C induces MYC expression in KRAS mutant non–small cell lung cancer (NSCLC) cells, an effect that can be suppressed by targeting MUC1-C via shRNA silencing, CRISPR editing, or pharmacologic inhibition with GO-203. MUC1-C activated the WNT/β-catenin (CTNNB1) pathway and promoted occupancy of MUC1-C/β-catenin/TCF4 complexes on the MYC promoter. MUC1-C also promoted the recruitment of the p300 histone acetylase (EP300) and, in turn, induced histone H3 acetylation and activation of MYC gene transcription. We also show that targeting MUC1-C decreased the expression of key MYC target genes essential for the growth and survival of NSCLC cells, such as TERT and CDK4. Based on these results, we found that the combination of GO-203 and the BET bromodomain inhibitor JQ1, which targets MYC transcription, synergistically suppressed MYC expression and cell survival in vitro as well as tumor xenograft growth. Furthermore, MUC1 expression significantly correlated with that of MYC and its target genes in human KRAS mutant NSCLC tumors. Taken together, these findings suggest a therapeutic approach for targeting MYC-dependent cancers and provide the framework for the ongoing clinical studies addressing the efficacy of MUC1-C inhibition in solid tumors. Cancer Res; 76(6); 1538–48. ©2016 AACR.
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- 2023
15. Supplementary Figure 1 from Inhibition of MUC1-C Suppresses MYC Expression and Attenuates Malignant Growth in KRAS Mutant Lung Adenocarcinomas
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Donald Kufe, Kwok-Kin Wong, Ashujit Tagde, Akriti Kharbanda, Maroof Alam, Caining Jin, Sean Pitroda, Hasan Rajabi, and Audrey Bouillez
- Abstract
Silencing MUC1-C suppresses MYC levels.
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- 2023
16. Supplementary Figure Legends from Inhibition of MUC1-C Suppresses MYC Expression and Attenuates Malignant Growth in KRAS Mutant Lung Adenocarcinomas
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Donald Kufe, Kwok-Kin Wong, Ashujit Tagde, Akriti Kharbanda, Maroof Alam, Caining Jin, Sean Pitroda, Hasan Rajabi, and Audrey Bouillez
- Abstract
Supplementary Figure Legends
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- 2023
17. Supplementary Figure 2 from Inhibition of MUC1-C Suppresses MYC Expression and Attenuates Malignant Growth in KRAS Mutant Lung Adenocarcinomas
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Donald Kufe, Kwok-Kin Wong, Ashujit Tagde, Akriti Kharbanda, Maroof Alam, Caining Jin, Sean Pitroda, Hasan Rajabi, and Audrey Bouillez
- Abstract
Effects of targeting MUC1-C in non-KRAS mutant NSCLC cells.
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- 2023
18. MUC1-C Induces PD-L1 and Immune Evasion in Triple-Negative Breast Cancer
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Donald Kufe, Maroof Alam, Ashujit Tagde, Hasan Rajabi, Xiufeng Hu, Tsuyoshi Hata, Yozo Suzuki, Masayuki Hiraki, Masaaki Miyo, Caining Jin, Takahiro Maeda, Kunihiko Hinohara, and Audrey Bouillez
- Subjects
0301 basic medicine ,Cancer Research ,Mice, Transgenic ,Triple Negative Breast Neoplasms ,Biology ,digestive system ,B7-H1 Antigen ,Article ,Epigenesis, Genetic ,GZMB ,03 medical and health sciences ,0302 clinical medicine ,Immune system ,Protein Domains ,Downregulation and upregulation ,Cell Line, Tumor ,PD-L1 ,Animals ,Humans ,skin and connective tissue diseases ,neoplasms ,Triple-negative breast cancer ,MUC1 ,Mucin-1 ,NF-kappa B ,Xenograft Model Antitumor Assays ,biological factors ,digestive system diseases ,Immune checkpoint ,Gene Expression Regulation, Neoplastic ,Protein Subunits ,030104 developmental biology ,Oncology ,030220 oncology & carcinogenesis ,Neoplastic Stem Cells ,Cancer research ,biology.protein ,Female ,Tumor Escape ,CD8 - Abstract
The immune checkpoint ligand PD-L1 and the transmembrane mucin MUC1 are upregulated in triple-negative breast cancer (TNBC), where they contribute to its aggressive pathogenesis. Here, we report that genetic or pharmacological targeting of the oncogenic MUC1 subunit MUC1-C is sufficient to suppress PD-L1 expression in TNBC cells. Mechanistic investigations showed that MUC1-C acted to elevate PD-L1 transcription by recruitment of MYC and NF-κB p65 to the PD-L1 promoter. In an immunocompetent model of TNBC in which Eo771/MUC1-C cells were engrafted into MUC1 transgenic mice, we showed that targeting MUC1-C associated with PD-L1 suppression, increases in tumor-infiltrating CD8+ T cells and tumor cell killing. MUC1 expression in TNBCs also correlated inversely with CD8, CD69, and GZMB, and downregulation of these markers associated with decreased survival. Taken together, our findings show how MUC1 contributes to immune escape in TNBC, and they offer a rationale to target MUC1-C as a novel immunotherapeutic approach for TNBC treatment. Significance: These findings show how upregulation of the transmembrane mucin MUC1 contributes to immune escape in an aggressive form of breast cancer, with potential implications for a novel immunotherapeutic approach. Cancer Res; 78(1); 205–15. ©2017 AACR.
- Published
- 2018
19. MUC1-C integrates PD-L1 induction with repression of immune effectors in non-small-cell lung cancer
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Donald Kufe, Ashujit Tagde, Hasan Rajabi, Surender Kharbanda, Takahiro Maeda, Mehmet Kemal Samur, Dennis Adeegbe, Masayuki Hiraki, Maroof Alam, Caining Jin, Kwok-Kin Wong, Audrey Bouillez, and Xiufeng Hu
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PD-L1 ,0301 basic medicine ,Cancer Research ,Lung Neoplasms ,Mice, Nude ,NSCLC ,Molecular oncology ,Article ,B7-H1 Antigen ,TLR9 ,Mice ,03 medical and health sciences ,Immune system ,Growth factor receptor ,Carcinoma, Non-Small-Cell Lung ,Cell Line, Tumor ,Genetics ,Animals ,Humans ,IFN-γ ,neoplasms ,Molecular Biology ,MUC1 ,TLR7 ,Immunity, Cellular ,biology ,Mucin-1 ,GM-CSF ,Acquired immune system ,respiratory tract diseases ,3. Good health ,Gene Expression Regulation, Neoplastic ,030104 developmental biology ,A549 Cells ,Cancer research ,biology.protein ,Female ,Tumor Escape ,MUC1-C ,MCP-1 ,Signal Transduction - Abstract
Immunotherapeutic approaches, particularly programmed death 1/programmed death ligand 1 (PD-1/PD-L1) blockade, have improved the treatment of non-small-cell lung cancer (NSCLC), supporting the premise that evasion of immune destruction is of importance for NSCLC progression. However, the signals responsible for upregulation of PD-L1 in NSCLC cells and whether they are integrated with the regulation of other immune-related genes are not known. Mucin 1 (MUC1) is aberrantly overexpressed in NSCLC, activates the nuclear factor-κB (NF-κB) p65→︀ZEB1 pathway and confers a poor prognosis. The present studies demonstrate that MUC1-C activates PD-L1 expression in NSCLC cells. We show that MUC1-C increases NF-κB p65 occupancy on the CD274/PD-L1 promoter and thereby drives CD274 transcription. Moreover, we demonstrate that MUC1-C-induced activation of NF-κB→︀ZEB1 signaling represses the TLR9 (toll-like receptor 9), IFNG, MCP-1 (monocyte chemoattractant protein-1) and GM-CSF genes, and that this signature is associated with decreases in overall survival. In concert with these results, targeting MUC1-C in NSCLC tumors suppresses PD-L1 and induces these effectors of innate and adaptive immunity. These findings support a previously unrecognized central role for MUC1-C in integrating PD-L1 activation with suppression of immune effectors and poor clinical outcome.
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- 2017
20. MUC1-C Represses the Crumbs Complex Polarity Factor CRB3 and Downregulates the Hippo Pathway
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Maroof Alam, Yozo Suzuki, Audrey Bouillez, Takahiro Maeda, Ashujit Tagde, Donald Kufe, Rehan Ahmad, Hasan Rajabi, and Masayuki Hiraki
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0301 basic medicine ,Cancer Research ,Down-Regulation ,Apoptosis ,Triple Negative Breast Neoplasms ,Protein Serine-Threonine Kinases ,Biology ,Article ,03 medical and health sciences ,Cell Line, Tumor ,Cell polarity ,Humans ,Hippo Signaling Pathway ,Phosphorylation ,Promoter Regions, Genetic ,skin and connective tissue diseases ,neoplasms ,Molecular Biology ,Transcription factor ,Adaptor Proteins, Signal Transducing ,Regulation of gene expression ,Hippo signaling pathway ,Membrane Glycoproteins ,Kinase ,Mucin-1 ,Wnt signaling pathway ,Cell Polarity ,Zinc Finger E-box-Binding Homeobox 1 ,Signal transducing adaptor protein ,YAP-Signaling Proteins ,Phosphoproteins ,digestive system diseases ,Cell biology ,Gene Expression Regulation, Neoplastic ,030104 developmental biology ,Oncology ,MCF-7 Cells ,Cancer research ,Female ,Signal transduction ,Signal Transduction ,Transcription Factors - Abstract
Apical–basal polarity and epithelial integrity are maintained in part by the Crumbs (CRB) complex. The C--terminal subunit of MUC1 (MUC1-C) is a transmembrane protein that is expressed at the apical border of normal epithelial cells and aberrantly at high levels over the entire surface of their transformed counterparts. However, it is not known whether MUC1-C contributes to this loss of polarity that is characteristic of carcinoma cells. Here it is demonstrated that MUC1-C downregulates expression of the Crumbs complex CRB3 protein in triple-negative breast cancer (TNBC) cells. MUC1-C associates with ZEB1 on the CRB3 promoter and represses CRB3 transcription. Notably, CRB3 activates the core kinase cassette of the Hippo pathway, which includes LATS1 and LATS2. In this context, targeting MUC1-C was associated with increased phosphorylation of LATS1, consistent with activation of the Hippo pathway, which is critical for regulating cell contact, tissue repair, proliferation, and apoptosis. Also shown is that MUC1-C--mediated suppression of CRB3 and the Hippo pathway is associated with dephosphorylation and activation of the oncogenic YAP protein. In turn, MUC1-C interacts with YAP, promotes formation of YAP/β-catenin complexes, and induces the WNT target gene MYC. These data support a previously unrecognized pathway in which targeting MUC1-C in TNBC cells (i) induces CRB3 expression, (ii) activates the CRB3-driven Hippo pathway, (iii) inactivates YAP, and thereby (iv) suppresses YAP/β-catenin–mediated induction of MYC expression. Implications: These findings demonstrate a previously unrecognized role for the MUC1-C oncoprotein in the regulation of polarity and the Hippo pathway in breast cancer. Mol Cancer Res; 14(12); 1266–76. ©2016 AACR.
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- 2016
21. MUC1-C activates BMI1 in human cancer cells
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Donald Kufe, Tahireh Markert, Maroof Alam, Kazumasa Komura, Kunihiko Hinohara, Yozo Suzuki, Masaaki Miyo, Rehan Ahmad, Masayuki Hiraki, Hasan Rajabi, Takahiro Maeda, Ashujit Tagde, and Audrey Bouillez
- Subjects
0301 basic medicine ,Cancer Research ,Transcription, Genetic ,Amino Acid Motifs ,H2A ,macromolecular substances ,Biology ,digestive system ,Article ,HOXC5 ,Histones ,Proto-Oncogene Proteins c-myc ,03 medical and health sciences ,CDKN2A ,Cell Line, Tumor ,Neoplasms ,Gene expression ,Genetics ,Humans ,Gene silencing ,Protein Interaction Domains and Motifs ,Amino Acid Sequence ,Gene Silencing ,Epigenetics ,Promoter Regions, Genetic ,skin and connective tissue diseases ,neoplasms ,Molecular Biology ,Cyclin-Dependent Kinase Inhibitor p16 ,MUC1 ,Homeodomain Proteins ,Polycomb Repressive Complex 1 ,Regulation of gene expression ,Mucin-1 ,NF-kappa B ,Ubiquitination ,HOXC13 ,BMI1 ,PRC1 ,p16INK4a tumor suppressor ,digestive system diseases ,3. Good health ,Gene Expression Regulation, Neoplastic ,030104 developmental biology ,Cancer cell ,Cancer research ,MUC1-C ,Protein Binding - Abstract
B-cell-specific Moloney murine leukemia virus integration site 1 (BMI1) is a component of the polycomb repressive complex 1 (PRC1) complex that is overexpressed in breast and other cancers, and promotes self-renewal of cancer stem-like cells. The oncogenic mucin 1 (MUC1) C-terminal (MUC1-C) subunit is similarly overexpressed in human carcinoma cells and has been linked to their self-renewal. There is no known relationship between MUC1-C and BMI1 in cancer. The present studies demonstrate that MUC1-C drives BMI1 transcription by a MYC-dependent mechanism in breast and other cancer cells. In addition, we show that MUC1-C blocks miR-200c-mediated downregulation of BMI1 expression. The functional significance of this MUC1-C→BMI1 pathway is supported by the demonstration that targeting MUC1-C suppresses BMI1-induced ubiquitylation of H2A and thereby derepresses homeobox HOXC5 and HOXC13 gene expression. Notably, our results further show that MUC1-C binds directly to BMI1 and promotes occupancy of BMI1 on the CDKN2A promoter. In concert with BMI1-induced repression of the p16INK4a tumor suppressor, we found that targeting MUC1-C is associated with induction of p16INK4a expression. In support of these results, analysis of three gene expresssion data sets demonstrated highly significant correlations between MUC1-C and BMI1 in breast cancers. These findings uncover a previously unrecognized role for MUC1-C in driving BMI1 expression and in directly interacting with this stem cell factor, linking MUC1-C with function of the PRC1 in epigenetic gene silencing.
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- 2016
22. DNA methylation by DNMT1 and DNMT3b methyltransferases is driven by the MUC1-C oncoprotein in human carcinoma cells
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Donald Kufe, Yozo Suzuki, Ashujit Tagde, Sean P. Pitroda, Audrey Bouillez, Maroof Alam, and Hasan Rajabi
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DNA (Cytosine-5-)-Methyltransferase 1 ,0301 basic medicine ,Cancer Research ,Epithelial-Mesenchymal Transition ,Methyltransferase ,Biology ,digestive system ,Article ,epigenetic regulation ,03 medical and health sciences ,Epigenetics of physical exercise ,Neoplasms ,Histone methylation ,Genetics ,Humans ,DNMT3b ,DNA (Cytosine-5-)-Methyltransferases ,Epigenetics ,Cancer epigenetics ,skin and connective tissue diseases ,neoplasms ,Molecular Biology ,RNA-Directed DNA Methylation ,Epigenomics ,DNA methylation ,Mucin-1 ,DNMT1 ,Transcription Factor RelA ,E-cadherin ,digestive system diseases ,3. Good health ,030104 developmental biology ,embryonic structures ,MCF-7 Cells ,Cancer research ,MUC1-C - Abstract
Aberrant expression of the DNA methyltransferases (DNMTs) and disruption of DNA methylation patterns are associated with carcinogenesis and cancer cell survival. The oncogenic MUC1-C protein is aberrantly overexpressed in diverse carcinomas; however, there is no known link between MUC1-C and DNA methylation. Our results demonstrate that MUC1-C induces the expression of DNMT1 and DNMT3b, but not DNMT3a, in breast and other carcinoma cell types. We show that MUC1-C occupies the DNMT1 and DNMT3b promoters in complexes with NF-κB p65 and drives DNMT1 and DNMT3b transcription. In this way, MUC1-C controls global DNA methylation as determined by analysis of LINE-1 repeat elements. The results further demonstrate that targeting MUC1-C downregulates DNA methylation of the CDH1 tumor suppressor gene in association with induction of E-cadherin expression. These findings provide compelling evidence that MUC1-C is of functional importance to induction of DNMT1 and DNMT3b and, in turn, changes in DNA methylation patterns in cancer cells.
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- 2016
23. Targeting MUC1-C suppresses BCL2A1 in triple-negative breast cancer
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Masayuki Hiraki, Ashujit Tagde, Maroof Alam, Audrey Bouillez, Donald Kufe, Surender Kharbanda, Caining Jin, Neha Mehrotra, Amy E. Keating, Tsuyoshi Hata, Takahiro Maeda, and Harpal Singh
- Subjects
0301 basic medicine ,Cancer Research ,lcsh:Medicine ,Drug resistance ,Article ,03 medical and health sciences ,0302 clinical medicine ,Breast cancer ,Downregulation and upregulation ,Genetics ,medicine ,lcsh:QH301-705.5 ,neoplasms ,Gene ,Triple-negative breast cancer ,MUC1 ,Chemistry ,lcsh:R ,medicine.disease ,3. Good health ,030104 developmental biology ,lcsh:Biology (General) ,Drug development ,030220 oncology & carcinogenesis ,Cancer research ,BCL2-related protein A1 - Abstract
B-cell lymphoma 2-related protein A1 (BCL2A1) is a member of the BCL-2 family of anti-apoptotic proteins that confers resistance to treatment with anti-cancer drugs; however, there are presently no agents that target BCL2A1. The MUC1-C oncoprotein is aberrantly expressed in triple-negative breast cancer (TNBC) cells, induces the epithelial–mesenchymal transition (EMT) and promotes anti-cancer drug resistance. The present study demonstrates that targeting MUC1-C genetically and pharmacologically in TNBC cells results in the downregulation of BCL2A1 expression. The results show that MUC1-C activates the BCL2A1 gene by an NF-κB p65-mediated mechanism, linking this pathway with the induction of EMT. The MCL-1 anti-apoptotic protein is also of importance for the survival of TNBC cells and is an attractive target for drug development. We found that inhibiting MCL-1 with the highly specific MS1 peptide results in the activation of the MUC1-C→NF-κB→BCL2A1 pathway. In addition, selection of TNBC cells for resistance to ABT-737, which inhibits BCL-2, BCL-xL and BCL-W but not MCL-1 or BCL2A1, is associated with the upregulation of MUC1-C and BCL2A1 expression. Targeting MUC1-C in ABT-737-resistant TNBC cells suppresses BCL2A1 and induces death, which is of potential therapeutic importance. These findings indicate that MUC1-C is a target for the treatment of TNBCs unresponsive to agents that inhibit anti-apoptotic members of the BCL-2 family., Treatment opportunities for tricky tumors New insights into gene regulation reveal vulnerabilities in a particularly hard-to-treat form of breast cancer. Patients with the ‘triple-negative’ form of this disease have a particularly poor prognosis, and researchers led by Donald Kufe at the Dana-Farber Cancer Institute in Boston, USA recently investigated the role of the mucin 1 protein in such malignancies. Mucin 1 promotes the survival and aggressive growth of tumor cells, and the researchers identified multiple downstream genes that mediate these effects. Through these pathways, mucin 1 helps establish resistance to multiple different therapeutic agents. However, Kufe’s team determined that GO-203, a drug currently in early clinical development, can effectively shut down these mucin 1-activated signaling pathways in cultured breast cancer cells. The researchers are now moving this agent into clinical trials to see whether triple-negative patients might benefit.
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- 2018
24. MUC1-C induces DNA methyltransferase 1 and represses tumor suppressor genes in acute myeloid leukemia
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David Avigan, Reddy Gali, Ashujit Tagde, Audrey Bouillez, Richard Stone, Donald Kufe, Dina Stroopinsky, Hasan Rajabi, Surender Kharbanda, and Maroof Alam
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0301 basic medicine ,Antigens, CD34 ,0302 clinical medicine ,hemic and lymphatic diseases ,Genes, Tumor Suppressor ,RNA, Small Interfering ,skin and connective tissue diseases ,Promoter Regions, Genetic ,education.field_of_study ,DNA methylation ,Gene Expression Regulation, Leukemic ,Myeloid leukemia ,Drug Synergism ,Cadherins ,3. Good health ,Leukemia, Myeloid, Acute ,Oncology ,030220 oncology & carcinogenesis ,embryonic structures ,Azacitidine ,Reprogramming ,medicine.drug ,DNA (Cytosine-5-)-Methyltransferase 1 ,Cell Survival ,Population ,Decitabine ,Biology ,DNA methyltransferase ,03 medical and health sciences ,Antigens, CD ,Cell Line, Tumor ,medicine ,PTEN ,Humans ,Gene Silencing ,education ,neoplasms ,CDH1 ,Mucin-1 ,DNMT1 ,Transcription Factor RelA ,Computational Biology ,NF-kappa B p50 Subunit ,ADP-ribosyl Cyclase 1 ,digestive system diseases ,030104 developmental biology ,Cancer research ,biology.protein ,MUC1-C ,Priority Research Paper - Abstract
// Ashujit Tagde 1 , Hasan Rajabi 1 , Dina Stroopinsky 2 , Reddy Gali 3 , Maroof Alam 1 , Audrey Bouillez 1 , Surender Kharbanda 1 , Richard Stone 1 , David Avigan 2 and Donald Kufe 1 1 Department of Medical Oncology, Dana-Farber Cancer Institute, Harvard Medical School, Boston, MA, USA 2 Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, MA, USA 3 Department of Biomedical Informatics, Harvard Medical School, Boston, MA, USA Correspondence to: David Avigan, email: // Donald Kufe, email: // Keywords : MUC1-C, DNMT1, DNA methylation, decitabine, CDH1 Received : April 25, 2016 Accepted : May 22, 2016 Published : June 01, 2016 Abstract Aberrant DNA methylation is a hallmark of acute myeloid leukemia (AML); however, the regulation of DNA methyltransferase 1 (DNMT1), which is responsible for maintenance of DNA methylation patterns, has largely remained elusive. MUC1-C is a transmembrane oncoprotein that is aberrantly expressed in AML stem-like cells. The present studies demonstrate that targeting MUC1-C with silencing or a pharmacologic inhibitor GO-203 suppresses DNMT1 expression. In addition, MUC1 expression positively correlates with that of DNMT1 in primary AML cells, particularly the CD34+/CD38- population. The mechanistic basis for this relationship is supported by the demonstration that MUC1-C activates the NF-κB p65 pathway, promotes occupancy of the MUC1-C/NF-κB complex on the DNMT1 promoter and drives DNMT1 transcription. We also show that targeting MUC1-C substantially reduces gene promoter-specific DNA methylation, and derepresses expression of tumor suppressor genes, including CDH1 , PTEN and BRCA1 . In support of these results, we demonstrate that combining GO-203 with the DNMT1 inhibitor decitabine is highly effective in reducing DNMT1 levels and decreasing AML cell survival. These findings indicate that (i) MUC1-C is an attractive target for the epigentic reprogramming of AML cells, and (ii) targeting MUC1-C in combination with decitabine is a potentially effective clinical approach for the treatment of AML.
- Published
- 2016
25. Unexpected Tricovalent Binding Mode of Boronic Acids within the Active Site of a Penicillin-Binding Protein
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R. F. Pratt, Bernard Joris, Fabio Prati, André Luxen, Eric Sauvage, Paulette Charlier, Astrid Zervosen, Frédéric Kerff, André Bouillez, Alexandre Herman, Raphaël Herman, and Jean-Marie Frère
- Subjects
Models, Molecular ,inorganic chemicals ,Boronic acid ,penicillin binding protein ,Crystal structure ,Stereochemistry ,Crystallography, X-Ray ,Biochemistry ,Catalysis ,Adduct ,Serine ,chemistry.chemical_compound ,Colloid and Surface Chemistry ,Nucleophile ,Catalytic Domain ,Hydrolase ,Penicillin-Binding Proteins ,Protease Inhibitors ,biology ,Chemistry ,Pinacol ,Active site ,General Chemistry ,Boronic Acids ,Serine-Type D-Ala-D-Ala Carboxypeptidase ,Kinetics ,Solvents ,biology.protein ,Amine gas treating ,Protein Binding - Abstract
Boronic acids bearing appropriate side chains are good inhibitors of serine amidohydrolases. The boron usually adopts a tetrahedral conformation, bound to the nucleophilic serine of the active site and mimicking the transition state of the enzymatic reaction. We have solved the structures of complexes of a penicillin-binding protein, the DD-peptidase from Actinomadura sp. R39, with four amidomethylboronic acids (2,6-dimethoxybenzamidomethylboronic acid, phenylacetamidomethylboronic acid, 2-chlorobenzamidomethylboronic acid, and 2-nitrobenzamidomethylboronic acid) and the pinacol ester derived from phenylacetamidomethylboronic acid. We found that, in each case, the boron forms a tricovalent adduct with Oγ of Ser49, Ser298, and the terminal amine group of Lys410, three key residues involved in the catalytic mechanism of penicillin-binding proteins. This represents the first tricovalent enzyme-inhibitor adducts observed by crystallography. In two of the five R39-boronate structures, the boronic acid is found as a tricovalent adduct in two monomers of the asymmetric unit and as a monocovalent adduct with the active serine in the two remaining monomers of the asymmetric unit. Formation of the tricovalent complex from a classical monocovalent complex may involve rotation around the Ser49 Cα-Cβ bond to place the boron in a position to interact with Ser298 and Lys410, and a twisting of the side-chain amide such that its carbonyl oxygen is able to hydrogen bond to the oxyanion hole NH of Thr413. Biphasic kinetics were observed in three of the five cases, and details of the reaction between R39 and 2,6-dimethoxybenzamidomethylboronic acid were studied. Observation of biphasic kinetics was not, however, thought to be correlated to formation of tricovalent complexes, assuming that the latter do form in solution. On the basis of the crystallographic and kinetic results, a reaction scheme for this unexpected inhibition by boronic acids is proposed.
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- 2011
26. MUC1-C drives MYC in multiple myeloma
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Maroof Alam, Donald Kufe, Audrey Bouillez, Teru Hideshima, David Avigan, Reddy Gali, Yu-Tzu Tai, Kenneth C. Anderson, Hasan Rajabi, Ashujit Tagde, and Shannon T. Bailey
- Subjects
0301 basic medicine ,Telomerase ,Transcription, Genetic ,Glutamate-Cysteine Ligase ,Immunology ,Down-Regulation ,Biology ,Response Elements ,Biochemistry ,digestive system ,Proto-Oncogene Proteins c-myc ,03 medical and health sciences ,0302 clinical medicine ,Cyclin D2 ,Transcription (biology) ,Cell Line, Tumor ,Gene silencing ,Humans ,skin and connective tissue diseases ,Transcription factor ,neoplasms ,beta Catenin ,Oligonucleotide Array Sequence Analysis ,Regulation of gene expression ,Messenger RNA ,Lymphoid Neoplasia ,Mucin-1 ,Cell Biology ,Hematology ,TCF4 ,Molecular biology ,biological factors ,digestive system diseases ,Gene Expression Regulation, Neoplastic ,030104 developmental biology ,030220 oncology & carcinogenesis ,CRISPR-Cas Systems ,Multiple Myeloma - Abstract
Multiple myeloma (MM) cell lines and primary tumor cells are addicted to the MYC oncoprotein for survival. Little is known, however, about how MYC expression is upregulated in MM cells. The mucin 1 C-terminal subunit (MUC1-C) is an oncogenic transmembrane protein that is aberrantly expressed in MM cell lines and primary tumor samples. The present studies demonstrate that targeting MUC1-C with silencing by clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated protein 9 editing or with the GO-203 inhibitor is associated with downregulation of MYC messenger RNA and protein. The results show that MUC1-C occupies the MYC promoter and thereby activates the MYC gene by a β-catenin/transcription factor 4 (TCF4)-mediated mechanism. In this way, MUC1-C (1) increases β-catenin occupancy on the MYC promoter, (2) forms a complex with β-catenin and TCF4, and, in turn, (3) drives MYC transcription. Analysis of MM cells using quantitative real-time reverse transcription polymerase chain reaction arrays further demonstrated that silencing MUC1-C is associated with downregulation of MYC target genes, including CCND2, hTERT, and GCLC Analysis of microarray data sets further demonstrated that MUC1 levels positively correlate with MYC expression in MM progression and in primary cells from over 800 MM patients. These findings collectively provide convincing evidence that MUC1-C drives MYC expression in MM.
- Published
- 2015
27. Inhibition of MUC1-C Suppresses MYC Expression and Attenuates Malignant Growth in KRAS Mutant Lung Adenocarcinomas
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Ashujit Tagde, Akriti Kharbanda, Audrey Bouillez, Maroof Alam, Kwok-Kin Wong, Sean P. Pitroda, Caining Jin, Donald Kufe, and Hasan Rajabi
- Subjects
0301 basic medicine ,Cancer Research ,Lung Neoplasms ,Cell Survival ,Mice, Nude ,Adenocarcinoma of Lung ,Biology ,Adenocarcinoma ,medicine.disease_cause ,Article ,Cell Line ,Small hairpin RNA ,Proto-Oncogene Proteins c-myc ,Proto-Oncogene Proteins p21(ras) ,03 medical and health sciences ,0302 clinical medicine ,Carcinoma, Non-Small-Cell Lung ,Cell Line, Tumor ,medicine ,Gene silencing ,Animals ,Humans ,RNA, Small Interfering ,Histone H3 acetylation ,EP300 ,neoplasms ,beta Catenin ,Mucin-1 ,Wnt signaling pathway ,TCF4 ,digestive system diseases ,Gene Expression Regulation, Neoplastic ,Wnt Proteins ,030104 developmental biology ,HEK293 Cells ,Oncology ,030220 oncology & carcinogenesis ,Mutation ,Cancer research ,Female ,KRAS ,Signal transduction ,Peptides ,E1A-Associated p300 Protein ,Signal Transduction - Abstract
Dysregulation of MYC expression is a hallmark of cancer, but the development of agents that target MYC has remained challenging. The oncogenic MUC1-C transmembrane protein is, like MYC, aberrantly expressed in diverse human cancers. The present studies demonstrate that MUC1-C induces MYC expression in KRAS mutant non–small cell lung cancer (NSCLC) cells, an effect that can be suppressed by targeting MUC1-C via shRNA silencing, CRISPR editing, or pharmacologic inhibition with GO-203. MUC1-C activated the WNT/β-catenin (CTNNB1) pathway and promoted occupancy of MUC1-C/β-catenin/TCF4 complexes on the MYC promoter. MUC1-C also promoted the recruitment of the p300 histone acetylase (EP300) and, in turn, induced histone H3 acetylation and activation of MYC gene transcription. We also show that targeting MUC1-C decreased the expression of key MYC target genes essential for the growth and survival of NSCLC cells, such as TERT and CDK4. Based on these results, we found that the combination of GO-203 and the BET bromodomain inhibitor JQ1, which targets MYC transcription, synergistically suppressed MYC expression and cell survival in vitro as well as tumor xenograft growth. Furthermore, MUC1 expression significantly correlated with that of MYC and its target genes in human KRAS mutant NSCLC tumors. Taken together, these findings suggest a therapeutic approach for targeting MYC-dependent cancers and provide the framework for the ongoing clinical studies addressing the efficacy of MUC1-C inhibition in solid tumors. Cancer Res; 76(6); 1538–48. ©2016 AACR.
- Published
- 2015
28. Abstract 5047: The MUC1 membrane-bound mucin increases tumor cell properties and chemoresistance in renal clear cell carcinoma
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Brigitte Hémon, Justine Woszczyk, Michael Perrais, Jean-Baptiste Gibier, Kelly Gaudelot, Sébastien Aubert, Isabelle Van Seuningen, Audrey Bouillez, Mélanie Fanchon, and Viviane Gnemmi
- Subjects
Oncology ,Cancer Research ,medicine.medical_specialty ,Matrigel ,Cell growth ,Mucin ,Cancer ,Transfection ,Biology ,medicine.disease ,Cell culture ,Internal medicine ,Cancer cell ,Cancer research ,medicine ,MUC1 - Abstract
Introduction: MUC1, an O-glycoprotein membrane-bound mucin, is overexpressed in renal clear-cell carcinomas (CRCC) with correlation to two major prognostic factors, Tumor-Node-Metastasis stage and nuclear Fürhman grade. Previously, we have shown that (i) MUC1 was significantly overexpressed in metastatic CRCC vs non-metastatic CRCC and (ii) MUC1 is a target gene of HIF-1 transcription factor which is a part of the hypoxia pathway, the main renal carcinogenetic pathway. Furthermore, CRCC is highly resistant to common systemic chemotherapies. Material and method: To better understand the roles of MUC1 in CRCC, we used two renal cell lines expressing MUC1 (786-O cells) or not (ACHN cells). 786-O cells were stably transfected with shRNA targeting MUC1 while ACHN cells with full-length MUC1. Proliferation, drug resistance, migration and invasion properties were studied in vitro in the different cellular clones using MTS cell proliferation assay, wound healing assay and Boyden chambers coated with Matrigel, respectively. Signaling pathways were screened by proteome profiler and western blot. Results and discussion: We showed that MUC1 expression was associated with increased invasion and migration properties of renal carcinomatous cells and a decrease of cell-cell interactions. MUC1 overexpressing cells (i) expressed higher levels of anti-apoptotic factors and MDR genes involved in chemoresistance processes and (ii) were more resistant to chemotherapeutic drugs. Conclusion: Our results show that MUC1 plays a role in biological properties of renal cancer cells suggesting important function for this mucin in tumour progression and chemo-resistance. Our data confirm its potential as a therapeutic target in this type of cancer. Citation Format: Kelly Gaudelot, Viviane Gnemmi, Audrey Bouillez, Jean-Baptiste Gibier, Mélanie Fanchon, Justine Woszczyk, Brigitte Hémon, Isabelle Van Seuningen, Sébastien Aubert, Michael Perrais. The MUC1 membrane-bound mucin increases tumor cell properties and chemoresistance in renal clear cell carcinoma. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 5047.
- Published
- 2016
29. Isolation and characterization of a primary proximal tubular epithelial cell model from human kidney by CD10/CD13 double labeling
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Cynthia Van der Hauwaert, Xavier Leroy, Laurent Zini, Grégoire Savary, Patrice Maboudou, Viviane Gnemmi, Michael Perrais, Audrey Bouillez, Nicolas Pottier, Christelle Cauffiez, François Glowacki, and Sébastien Aubert
- Subjects
Pathology ,Anatomy and Physiology ,lcsh:Medicine ,Cell Separation ,Nephrectomy ,Kidney Tubules, Proximal ,0302 clinical medicine ,Molecular Cell Biology ,Electric Impedance ,lcsh:Science ,Cells, Cultured ,0303 health sciences ,Kidney ,education.field_of_study ,Multidisciplinary ,medicine.diagnostic_test ,Cell sorting ,Flow Cytometry ,medicine.anatomical_structure ,Phenotype ,Nephrology ,030220 oncology & carcinogenesis ,Medicine ,Neprilysin ,Cellular Types ,Research Article ,medicine.medical_specialty ,Cell type ,Cell Physiology ,Population ,Primary Cell Culture ,Biology ,CD13 Antigens ,Flow cytometry ,03 medical and health sciences ,medicine ,Humans ,education ,Cell Shape ,030304 developmental biology ,Renal Physiology ,Staining and Labeling ,lcsh:R ,Epithelial Cells ,Renal System ,Epithelium ,Cell culture ,Renal physiology ,lcsh:Q ,Cytometry - Abstract
Renal proximal tubular epithelial cells play a central role in renal physiology and are among the cell types most sensitive to ischemia and xenobiotic nephrotoxicity. In order to investigate the molecular and cellular mechanisms underlying the pathophysiology of kidney injuries, a stable and well-characterized primary culture model of proximal tubular cells is required. An existing model of proximal tubular cells is hampered by the cellular heterogeneity of kidney; a method based on cell sorting for specific markers must therefore be developed. In this study, we present a primary culture model based on the mechanical and enzymatic dissociation of healthy tissue obtained from nephrectomy specimens. Renal epithelial cells were sorted using co-labeling for CD10 and CD13, two renal proximal tubular epithelial markers, by flow cytometry. Their purity, phenotypic stability and functional properties were evaluated over several passages. Our results demonstrate that CD10/CD13 double-positive cells constitute a pure, functional and stable proximal tubular epithelial cell population that displays proximal tubule markers and epithelial characteristics over the long term, whereas cells positive for either CD10 or CD13 alone appear to be heterogeneous. In conclusion, this study describes a method for establishing a robust renal proximal tubular epithelial cell model suitable for further experimentation.
- Published
- 2012
30. Synthesis and evaluation of boronic acids as inhibitors of Penicillin Binding Proteins of classes A, B and C
- Author
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Alexandre Herman, André Luxen, Ana Maria Amoroso, André Bouillez, Astrid Zervosen, Bernard Joris, Eric Sauvage, and Paulette Charlier
- Subjects
Penicillin binding proteins ,medicine.drug_class ,Clinical Biochemistry ,Antibiotics ,Pharmaceutical Science ,medicine.disease_cause ,Biochemistry ,Microbiology ,Structure-Activity Relationship ,Antibiotic resistance ,Drug Discovery ,Streptococcus pneumoniae ,Actinomycetales ,polycyclic compounds ,medicine ,Penicillin-Binding Proteins ,Actinomadura ,Molecular Biology ,chemistry.chemical_classification ,biology ,Dose-Response Relationship, Drug ,Molecular Structure ,Chemistry ,Organic Chemistry ,biochemical phenomena, metabolism, and nutrition ,biology.organism_classification ,Boronic Acids ,Penicillin ,Enzyme ,Molecular Medicine ,Bacteria ,medicine.drug - Abstract
In response to the widespread use of β-lactam antibiotics bacteria have evolved drug resistance mechanisms that include the production of resistant Penicillin Binding Proteins (PBPs). Boronic acids are potent β-lactamase inhibitors and have been shown to display some specificity for soluble transpeptidases and PBPs, but their potential as inhibitors of the latter enzymes is yet to be widely explored. Recently, a (2,6-dimethoxybenzamido)methylboronic acid was identified as being a potent inhibitor of Actinomadura sp. R39 transpeptidase (IC50: 1.3 μM). In this work, we synthesized and studied the potential of a number of acylaminomethylboronic acids as inhibitors of PBPs from different classes. Several derivatives inhibited PBPs of classes A, B and C from penicillin sensitive strains. The (2-nitrobenzamido)methylboronic acid was identified as a good inhibitor of a class A PBP (PBP1b from Streptococcus pneumoniae, IC50 = 26 μM), a class B PBP (PBP2xR6 from Streptococcus pneumoniae, IC50 = 138 μM) and a class C PBP (R39 from Actinomadura sp., IC50 = 0.6 μM). This work opens new avenues towards the development of molecules that inhibit PBPs, and eventually display bactericidal effects, on distinct bacterial species.
- Published
- 2012
31. Protected Silver Coating for NIF Flashlamp Reflectors*
- Author
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Norman Thomas, Jesse Wolfe, Joseph Farmer, Al Erlandson, Chris Marshall, Dan Woodruff, Dan Bouillez, Darin Glenn, and Erik Bjornard
- Abstract
The National Ignition Facility at Lawrence Livermore National Laboratory will be a national center to study inertial confinement fusion and the physics of high energy and pressure. The $1.2 billion project consists of 192 laser beams which will direct more than 500 trillion watts towards a tiny target in a pulse only a billionth of a second long. The laser contains neodymium glass amplifiers which are pumped by xenon flashlamps. The flashlamps have silver reflectors to direct the light toward the glass laser slabs. Silver is ideal because of its high reflectance from 400 to 1000 nm wavelength which matches the neodymium absorption cross-section. In order to maintain the overall efficiency of the laser beams, there is an effort to prevent the corrosion of the silver reflectors by various means. One solution is the use of durable protected silver coatings which must survive 24,000 exposures to intense flashlamp light over a 30-year period while being exposed to small levels of atmospheric corrodants. One protected silver reflector has survived 20,717 flashlamp discharges so far in 100% Livermore atmosphere with no change in reflectance. This paper describes another protected silver reflector which is very durable.
- Published
- 1997
32. 344 The MUC1 Membrane-bound Mucin is an Actor in Renal Clear-cell Carcinoma
- Author
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Bélinda Ringot, Laurent Zini, Audrey Bouillez, A. Aubert, Michael Perrais, Caroline Butruille, Xavier Leroy, Viviane Gnemmi, and I. Van Seuningen
- Subjects
Oncology ,Renal clear cell carcinoma ,Cancer Research ,medicine.medical_specialty ,Membrane bound ,Chemistry ,Internal medicine ,Mucin ,Cancer research ,medicine ,MUC1 - Published
- 2012
33. Abstract 2737: The MUC1 membrane-bound mucin is an actor in renal tumor progression
- Author
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Bélinda Ringot, Caroline Butruille, Audrey Bouillez, Brigitte Hémon, Michael Perrais, Laurent Zini, Sébastien Aubert, Xavier Leroy, Viviane Gnemmi, and Isabelle Van Seuningen
- Subjects
Cancer Research ,Pathology ,medicine.medical_specialty ,medicine.diagnostic_test ,Mucin ,Cancer ,Transfection ,Biology ,medicine.disease ,digestive system ,biological factors ,digestive system diseases ,Metastasis ,Flow cytometry ,Oncology ,Tumor progression ,Cancer cell ,Cancer research ,medicine ,skin and connective tissue diseases ,neoplasms ,MUC1 - Abstract
MUC1, an O-glycoprotein membrane-bound mucin, is overexpressed in clear-cell carcinomas (cRCC) with correlation to two prognostic factors, tumor-node metastasis stage and nuclear Fürhman grade. Previously (Aubert et al, Cancer Res 2009), we have shown that MUC1 was significantly overexpressed in metastatic cRCC vs non-metastatic cRCC and MUC1 is a target gene of HIF-1 transcription factor which is a part of the hypoxia pathway, the main renal carcinogenetic pathway. To better understand the roles of MUC1 in cRCC, we used two renal cell lines expressing MUC1 (Caki-2) or not (ACHN cells). Caki-2 cells were stably transfected with shRNA targeting MUC1 and ACHN cells with an expressing vector containing MUC1 cDNA. Flow cytometry assay and confocal microscopy showed a homogenous membrane expression of MUC1 but also in the nucleus. In vitro studies underlined MUC1 role in the invasion, migration and anchorage-independent growth. In contrast, MUC1 had no role in proliferation and clonogenicity. Cellular agreggation test showed that MUC1 expression is associated with a decrease of cell-cell interactions. Phospho-JNK and phospho-p38 levels were higher in MUC1 expressing cells. Transcriptomic analysis results performed on 44K Agilent Microarrays suggest that MUC1 is a candidate in chemoresistance. Subcutaneous xenograft will be also performed. Our results show that MUC1 plays a role in biological properties of renal cancer cells suggesting the important function for this mucin in tumor progression and confirms its potential as a therapeutic target in this type of cancer. 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 2737. doi:10.1158/1538-7445.AM2011-2737
- Published
- 2011
34. Abstract 5250: MUC1 expression is induced by Snai1 in an epithelial mesenchymal transition renal model
- Author
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Laurent Zini, Michael Perrais, Bélinda Ringot, Aubert Sébastien, Xavier Leroy, Audrey Bouillez, Isabelle Van-Seuningen, Marie-Christine Copin, and Viviane Gnemmi
- Subjects
Cancer Research ,Tissue microarray ,Mesenchymal stem cell ,Cancer ,Biology ,medicine.disease ,digestive system diseases ,Chromatin ,Oncology ,SNAI1 ,Cancer research ,medicine ,Epithelial–mesenchymal transition ,Sarcomatoid carcinoma ,neoplasms ,MUC1 - Abstract
Accumulating data suggest that epithelial mesenchymal transition (EMT) is involved in cancer progression. Sarcomatoid carcinoma represents a high-grade transformation and an EMT-like pattern. Up to 8% of conventional renal cells cancers (cRCC) present with sarcomatoid carcinoma component that portends a worse prognosis. MUC1, a membrane-bound glycoprotein is known to be over-expressed in cRCC with correlation to prognosis. Indeed, we showed that MUC1 overexpression is correlated with a metastatic phenotype using tissue microarray sampling 27 cRCC of same TNM stage. Also, we demonstrated in a series of 15 RCC with sarcomatoid component, the loss of epithelial markers and the acquisition of mesenchymal markers compared to conventional carcinomatous areas. MUC1 was significantly overexpressed along with Snai1, a major actor of EMT, in sarcomatoid component. So, we hypothesized that MUC1 could be regulated by Snai1. Bioinformatic analyses revealed two Snai1 E-boxes at -84/-72 on MUC1 promoter. Cotransfection studies of MUC1 promoter and Snai1 expression vectors in renal cells lines showed an induction of MUC1 transcriptional activity by luciferase assays. Mutagenesis site-directed of both Snai1 E-boxes in renal cells lines induced loss MUC1 transcriptional activity. By chromatin immunoprecipation assay and gel shift analysis we will demonstrate a direct interaction between Snai1 and MUC1 promoter. In conclusion, all these data revealed that MUC1, usually considered as an epithelial marker, is overexpressed during cancer linked EMT process with a direct regulation by Snai1 suggesting that MUC1 plays a role in renal cancer progression. 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 5250. doi:10.1158/1538-7445.AM2011-5250
- Published
- 2011
35. Abstract 2457: The MUC1 membrane-bound mucin is an actor in renal clear-cell carcinoma
- Author
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Brigitte Hémon, Laurent Zini, Viviane Gnemmi, Michael Perrais, Audrey Bouillez, Xavier Leroy, Sébastien Aubert, and Isabelle Van Seuningen
- Subjects
Cancer Research ,Matrigel ,Pathology ,medicine.medical_specialty ,Cell growth ,business.industry ,Cancer ,Transfection ,medicine.disease ,Oncology ,Cell culture ,Apoptosis ,Cancer cell ,medicine ,Cancer research ,business ,MUC1 - Abstract
MUC1, an O-glycoprotein membrane-bound mucin, is overexpressed in renal clear-cell carcinomas (CRCC) with correlation to two major prognostic factors, Tumor-Node-Metastasis stage and nuclear Fürhman grade. Previously (Aubert et al., Cancer Res 2009), we have shown that (i) MUC1 was significantly overexpressed in metastatic CRCC vs non-metastatic CRCC and (ii) MUC1 is a target gene of HIF-1 transcription factor which is a part of the hypoxia pathway, the main renal carcinogenetic pathway. To better understand the roles of MUC1 in CRCC, we used two renal cell lines expressing MUC1 (Caki-2 cells) or not (ACHN cells). Caki-2 cells were stably transfected with shRNA targeting MUC1 and ACHN cells with an expressing vector containing MUC1 cDNA. Proliferation, migration and invasion properties were studied in vitro in the different cellular clones using Boyden chambers coated with Matrigel, wound healing assay and MTS cell proliferation assay, respectively. Our results showed that (i) MUC1 expression was associated with an increase of invasion and migration properties of renal carcinoma cells whereas no effect on proliferation and apoptotic rate was observed and (ii) levels of phospho-JNK were higher in MUC1 expressing cells. Results of (i) transcriptomic analysis on 44K Agilent microarrays, (ii) signaling pathway screening and (iii) subcutaneous xenografts will be presented. Our results show that MUC1 plays a role in biological properties of renal cancer cells suggesting important function for this mucin in tumour progression and confirms its potential as a therapeutic target in this type of cancer. Note: This abstract was not presented at the AACR 101st Annual Meeting 2010 because the presenter was unable to attend. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 2457.
- Published
- 2010
36. Toward a novel metal based chemotherapy against tropical diseases 4. Synthesis and characterization of new metal-clotrimazole complexes and evaluation of their activity against Trypanosoma cruzi
- Author
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Dante Masi, Franklin Vargas, André Bouillez, Maribel Navarro, M. V. Capparelli, Reinaldo Atencio, Keyla Lazardi, Julio A. Urbina, Alfred F. Noels, and Roberto A. Sánchez-Delgado
- Subjects
biology ,Clotrimazole ,Chemistry ,Stereochemistry ,Ligand ,Triclinic crystal system ,biology.organism_classification ,Inorganic Chemistry ,Metal ,Crystallography ,chemistry.chemical_compound ,visual_art ,Materials Chemistry ,visual_art.visual_art_medium ,medicine ,Imidazole ,Orthorhombic crystal system ,Physical and Theoretical Chemistry ,Trypanosoma cruzi ,medicine.drug ,Monoclinic crystal system - Abstract
The syntheses and characterization of metal imidazole complexes showing activity against Trypanosoma cruzi, the causative agent of Chagas disease, are presented, RuCl2(CTZ)2 (2) and RuCl2(BTZ)2 (4) were prepared by reaction of RuCl2(NCCH3)4 (1) with the appropriate ligand (CTZ, clotrimazole = 1-[(2-chlorophenyl)diphenylmethyl]-1H-imidazole; BTZ = 1-[(2-bromophenyl)diphenylmethyl]-1H-imidazole) [Ru(bipy)(CTZ)2](PF6)2 (3) (bipy=2,2′-bipyridyl) was obtained by reaction of 2 with bipy and NH4PF6 in MeCN. Reaction of [RhCl(COD)]2 with CTZ yielded RhCl(COD)(CTZ) (5) (COD=1,5-cyclooctadiene), while AuCl1CTZ (6), K2[PtCl4(CTZ)2] (7) and [Cu(CTZ)2]PF6 (8) were obtained by interaction of CTZ with AuCl1·HCl, K2PtCl4 and [Cu(CH3CN)4]PF6, respectively. All the new complexes were characterized by NMR and other appropriate techniques, X-ray diffraction studies of 4·3H2O, 5 and 6 were also carried out. The structure of 4·3H2O consists of a distorted tetrahedral arrangement of two N atoms from the BTZ ligands and two Cl atoms around the Ru(II) ion; 4·3H2O crystallizes in the orthorhombic space group Puma (No. 62) with a=12.818(5), b=29.115(5), c=12.040(5) A , V=4493.2(8) A and Z=4. Complex 5 displayed a square planar structure typical for Rh(1) bound to N from CTZ, Cl, and the two CC bonds of COD; 5 crystallized in the triclinic space group (P(= l) (No. 2) with a=12.407(3), b=12.876(4), c=10.069(3) A , α=111.59(2)°, β=107.80(2)°, γ=103.28(2)°, V=1313.4(8) A 3 and Z=2. Complex 6 also displayed a square arrangement of N from CTZ, plus three Cl atoms around the Au(III) ion; 6 crystallized in the monoclinic space group P21/n (No. 14) with a=9.507(1), b=18.280(4), c=12.877(1) A , β=100.59(1)°, V=2199.7(5) A 3 and Z=4. All the new compounds were found to be active against in vitro cultures of Trypanosoma cruzi, following the trend 3 = 7
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