47 results on '"Vesuna F"'
Search Results
2. Interleukin-6 induces an epithelial–mesenchymal transition phenotype in human breast cancer cells
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Sullivan, N J, Sasser, A K, Axel, A E, Vesuna, F, Raman, V, Ramirez, N, Oberyszyn, T M, and Hall, B M
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- 2009
- Full Text
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3. Hypoxia-induced human endonuclease G expression suppresses tumor growth in a xenograft model
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Winnard, Jr, P T, Botlagunta, M, Kluth, J B, Mukadam, S, Krishnamachary, B, Vesuna, F, and Raman, V
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- 2008
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4. Oncogenic role of DDX3 in breast cancer biogenesis
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Botlagunta, M, Vesuna, F, Mironchik, Y, Raman, A, Lisok, A, Winnard, Jr, P, Mukadam, S, Van Diest, P, Chen, J H, Farabaugh, P, Patel, A H, and Raman, V
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- 2008
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5. Nuclear DDX3 expression predicts poor outcome in colorectal and breast cancer
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Heerma van Voss MR, Vesuna F, Bol GM, Meeldijk J, Raman A, Offerhaus GJ, Buerger H, Patel AH, van der Wall E, van Diest PJ, and Raman V
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RNA helicase ,DDX3X ,nuclear export ,lcsh:Neoplasms. Tumors. Oncology. Including cancer and carcinogens ,CRM1 ,lcsh:RC254-282 - Abstract
Marise R Heerma van Voss,1,2 Farhad Vesuna,2 Guus M Bol,1,2 Jan Meeldijk,1,3 Ana Raman,4 G Johan Offerhaus,1 Horst Buerger,5 Arvind H Patel,6 Elsken van der Wall,7,8 Paul J van Diest,1,8 Venu Raman1,2,8 1Department of Pathology, University Medical Center Utrecht, Utrecht, the Netherlands; 2Department of Radiology, Johns Hopkins University School of Medicine, Baltimore, MD, USA; 3Laboratory of Translational Immunology, University Medical Center Utrecht, Utrecht, the Netherlands; 4Department of Pharmacology, Johns Hopkins University School of Medicine, Baltimore, MD, USA; 5Institute of Pathology, Paderborn, Germany; 6Centre for Virus Research, MRC-University of Glasgow, Glasgow, UK; 7Cancer Center, University Medical Center Utrecht, Utrecht, the Netherlands; 8Department of Oncology, Johns Hopkins University School of Medicine, Baltimore, MD, USA Purpose: DEAD box protein 3 (DDX3) is an RNA helicase with oncogenic properties that shuttles between the cytoplasm and nucleus. The majority of DDX3 is found in the cytoplasm, but a subset of tumors has distinct nuclear DDX3 localization of yet unknown biological significance. This study aimed to evaluate the significance of and mechanisms behind nuclear DDX3 expression in colorectal and breast cancer.Methods: Expression of nuclear DDX3 and the nuclear exporter chromosome region maintenance 1 (CRM1) was evaluated by immunohistochemistry in 304 colorectal and 292 breast cancer patient samples. Correlations between the subcellular localization of DDX3 and CRM1 and the difference in overall survival between patients with and without nuclear DDX3 were studied. In addition, DDX3 mutants were created for in vitro evaluation of the mechanism behind nuclear retention of DDX3.Results: DDX3 was present in the nucleus of 35% of colorectal and 48% of breast cancer patient samples and was particularly strong in the nucleolus. Nuclear DDX3 correlated with worse overall survival in both colorectal (hazard ratio [HR] 2.34, P
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- 2017
6. Targeting mitochondrial translation by inhibiting DDX3 : A novel radiosensitization strategy for cancer treatment
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Heerma Van Voss, M. R., Vesuna, F., Bol, G. M., Afzal, J., Tantravedi, Saritha, Bergman, Y., Kammers, Kai, Lehar, M., Malek, Reem, Ballew, M., ter Hoeve, N., Abou, D., Thorek, D., Berlinicke, Cynthia, Yazdankhah, M., Sinha, D., le Couteur, A., Abrahams, R., Tran, Phuoc T., Van Diest, P. J., Raman, V., Heerma Van Voss, M. R., Vesuna, F., Bol, G. M., Afzal, J., Tantravedi, Saritha, Bergman, Y., Kammers, Kai, Lehar, M., Malek, Reem, Ballew, M., ter Hoeve, N., Abou, D., Thorek, D., Berlinicke, Cynthia, Yazdankhah, M., Sinha, D., le Couteur, A., Abrahams, R., Tran, Phuoc T., Van Diest, P. J., and Raman, V.
- Published
- 2018
7. Targeting mitochondrial translation by inhibiting DDX3: A novel radiosensitization strategy for cancer treatment
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Pathologie Groep Van Diest, Cancer, Unit Opleiding Aios, Pathologie, Heerma Van Voss, M. R., Vesuna, F., Bol, G. M., Afzal, J., Tantravedi, Saritha, Bergman, Y., Kammers, Kai, Lehar, M., Malek, Reem, Ballew, M., ter Hoeve, N., Abou, D., Thorek, D., Berlinicke, Cynthia, Yazdankhah, M., Sinha, D., le Couteur, A., Abrahams, R., Tran, Phuoc T., Van Diest, P. J., Raman, V., Pathologie Groep Van Diest, Cancer, Unit Opleiding Aios, Pathologie, Heerma Van Voss, M. R., Vesuna, F., Bol, G. M., Afzal, J., Tantravedi, Saritha, Bergman, Y., Kammers, Kai, Lehar, M., Malek, Reem, Ballew, M., ter Hoeve, N., Abou, D., Thorek, D., Berlinicke, Cynthia, Yazdankhah, M., Sinha, D., le Couteur, A., Abrahams, R., Tran, Phuoc T., Van Diest, P. J., and Raman, V.
- Published
- 2018
8. Targeting mitochondrial translation by inhibiting DDX3: a novel radiosensitization strategy for cancer treatment
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Heerma van Voss, M R, primary, Vesuna, F, additional, Bol, G M, additional, Afzal, J, additional, Tantravedi, S, additional, Bergman, Y, additional, Kammers, K, additional, Lehar, M, additional, Malek, R, additional, Ballew, M, additional, ter Hoeve, N, additional, Abou, D, additional, Thorek, D, additional, Berlinicke, C, additional, Yazdankhah, M, additional, Sinha, D, additional, Le, A, additional, Abrahams, R, additional, Tran, P T, additional, van Diest, P J, additional, and Raman, V, additional
- Published
- 2017
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9. Expression of DDX3 is directly modulated by hypoxia inducible factor-1 alpha in breast epithelial cells
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Botlagunta, M., Krishnamachary, B., Vesuna, F., Winnard, P.T., Bol, G.M., Patel, A.H., and Ramanathan, V.
- Abstract
DEAD box protein, DDX3, is aberrantly expressed in breast cancer cells ranging from weakly invasive to aggressive phenotypes and functions as an important regulator of cancer cell growth and survival. Here, we demonstrate that hypoxia inducible factor-1 alpha is a transcriptional activator of DDX3 in breast cancer cells. Within the promoter region of the human DDX3 gene, we identified three putative hypoxia inducible factor-1 responsive elements. By luciferase reporter assays in combination with mutated hypoxia inducible factor-1 responsive elements, we determined that the hypoxia inducible factor-1 responsive element at position -153 relative to the translation start site is essential for transcriptional activation of DDX3 under hypoxic conditions. We also demonstrated that hypoxia inducible factor-1 binds to the DDX3 promoter and that the binding is specific, as revealed by siRNA against hypoxia inducible factor-1 and chromatin immunoprecipitation assays. Thus, the activation of DDX3 expression during hypoxia is due to the direct binding of hypoxia inducible factor-1 to hypoxia responsive elements in the DDX3 promoter. In addition, we observed a significant overlap in the protein expression pattern of hypoxia inducible factor-1 alpha and DDX3 in MDA-MB-231 xenograft tumors. Taken together, our results demonstrate, for the first time, the role of DDX3 as a hypoxia-inducible gene that exhibits enhanced expression through the interaction of hypoxia inducible factor-1 with hypoxia inducible factor-1 responsive elements in its promoter region
- Published
- 2011
10. Targeting mitochondrial translation by inhibiting DDX3: a novel radiosensitization strategy for cancer treatment
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Heerma van Voss, M R, Vesuna, F, Bol, G M, Afzal, J, Tantravedi, S, Bergman, Y, Kammers, K, Lehar, M, Malek, R, Ballew, M, ter Hoeve, N, Abou, D, Thorek, D, Berlinicke, C, Yazdankhah, M, Sinha, D, Le, A, Abrahams, R, Tran, P T, van Diest, P J, and Raman, V
- Abstract
DDX3 is a DEAD box RNA helicase with oncogenic properties. RK-33 is developed as a small-molecule inhibitor of DDX3 and showed potent radiosensitizing activity in preclinical tumor models. This study aimed to assess DDX3 as a target in breast cancer and to elucidate how RK-33 exerts its anti-neoplastic effects. High DDX3 expression was present in 35% of breast cancer patient samples and correlated with markers of aggressiveness and shorter survival. With a quantitative proteomics approach, we identified proteins involved in the mitochondrial translation and respiratory electron transport pathways to be significantly downregulated after RK-33 or DDX3 knockdown. DDX3 localized to the mitochondria and DDX3 inhibition with RK-33 reduced mitochondrial translation. As a consequence, oxygen consumption rates and intracellular ATP concentrations decreased and reactive oxygen species (ROS) increased. RK-33 antagonized the increase in oxygen consumption and ATP production observed after exposure to ionizing radiation and reduced DNA repair. Overall, we conclude that DDX3 inhibition with RK-33 causes radiosensitization in breast cancer through inhibition of mitochondrial translation, which results in reduced oxidative phosphorylation capacity and increased ROS levels, culminating in a bioenergetic catastrophe.
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- 2018
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11. The Transcription Factor Encyclopedia
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Yusuf, D, Butland, SL, Swanson, MI, Bolotin, E, Ticoll, A, Cheung, WA, Zhang, XYC, Dickman, CTD, Fulton, DL, Lim, JS, Schnabl, JM, Ramos, OHP, Vasseur-Cognet, M, de Leeuw, CN, Simpson, EM, Ryffel, GU, Lam, EW-F, Kist, R, Wilson, MSC, Marco-Ferreres, R, Brosens, JJ, Beccari, LL, Bovolenta, P, Benayoun, BA, Monteiro, LJ, Schwenen, HDC, Grontved, L, Wederell, E, Mandrup, S, Veitia, RA, Chakravarthy, H, Hoodless, PA, Mancarelli, MM, Torbett, BE, Banham, AH, Reddy, SP, Cullum, RL, Liedtke, M, Tschan, MP, Vaz, M, Rizzino, A, Zannini, M, Frietze, S, Farnham, PJ, Eijkelenboom, A, Brown, PJ, Laperriere, D, Leprince, D, de Cristofaro, T, Prince, KL, Putker, M, del Peso, L, Camenisch, G, Wenger, RH, Mikula, M, Rozendaal, M, Mader, S, Ostrowski, J, Rhodes, SJ, Van Rechem, C, Boulay, G, Olechnowicz, SWZ, Breslin, MB, Lan, MS, Nanan, KK, Wegner, M, Hou, J, Mullen, RD, Colvin, SC, Noy, PJ, Webb, CF, Witek, ME, Ferrell, S, Daniel, JM, Park, J, Waldman, SA, Peet, DJ, Taggart, M, Jayaraman, P-S, Karrich, JJ, Blom, B, Vesuna, F, O'Geen, H, Sun, Y, Gronostajski, RM, Woodcroft, MW, Hough, MR, Chen, E, Europe-Finner, GN, Karolczak-Bayatti, M, Bailey, J, Hankinson, O, Raman, V, LeBrun, DP, Biswal, S, Harvey, CJ, DeBruyne, JP, Hogenesch, JB, Hevner, RF, Heligon, C, Luo, XM, Blank, MC, Millen, KJ, Sharlin, DS, Forrest, D, Dahlman-Wright, K, Zhao, C, Mishima, Y, Sinha, S, Chakrabarti, R, Portales-Casamar, E, Sladek, FM, Bradley, PH, Wasserman, WW, Yusuf, D, Butland, SL, Swanson, MI, Bolotin, E, Ticoll, A, Cheung, WA, Zhang, XYC, Dickman, CTD, Fulton, DL, Lim, JS, Schnabl, JM, Ramos, OHP, Vasseur-Cognet, M, de Leeuw, CN, Simpson, EM, Ryffel, GU, Lam, EW-F, Kist, R, Wilson, MSC, Marco-Ferreres, R, Brosens, JJ, Beccari, LL, Bovolenta, P, Benayoun, BA, Monteiro, LJ, Schwenen, HDC, Grontved, L, Wederell, E, Mandrup, S, Veitia, RA, Chakravarthy, H, Hoodless, PA, Mancarelli, MM, Torbett, BE, Banham, AH, Reddy, SP, Cullum, RL, Liedtke, M, Tschan, MP, Vaz, M, Rizzino, A, Zannini, M, Frietze, S, Farnham, PJ, Eijkelenboom, A, Brown, PJ, Laperriere, D, Leprince, D, de Cristofaro, T, Prince, KL, Putker, M, del Peso, L, Camenisch, G, Wenger, RH, Mikula, M, Rozendaal, M, Mader, S, Ostrowski, J, Rhodes, SJ, Van Rechem, C, Boulay, G, Olechnowicz, SWZ, Breslin, MB, Lan, MS, Nanan, KK, Wegner, M, Hou, J, Mullen, RD, Colvin, SC, Noy, PJ, Webb, CF, Witek, ME, Ferrell, S, Daniel, JM, Park, J, Waldman, SA, Peet, DJ, Taggart, M, Jayaraman, P-S, Karrich, JJ, Blom, B, Vesuna, F, O'Geen, H, Sun, Y, Gronostajski, RM, Woodcroft, MW, Hough, MR, Chen, E, Europe-Finner, GN, Karolczak-Bayatti, M, Bailey, J, Hankinson, O, Raman, V, LeBrun, DP, Biswal, S, Harvey, CJ, DeBruyne, JP, Hogenesch, JB, Hevner, RF, Heligon, C, Luo, XM, Blank, MC, Millen, KJ, Sharlin, DS, Forrest, D, Dahlman-Wright, K, Zhao, C, Mishima, Y, Sinha, S, Chakrabarti, R, Portales-Casamar, E, Sladek, FM, Bradley, PH, and Wasserman, WW
- Abstract
Here we present the Transcription Factor Encyclopedia (TFe), a new web-based compendium of mini review articles on transcription factors (TFs) that is founded on the principles of open access and collaboration. Our consortium of over 100 researchers has collectively contributed over 130 mini review articles on pertinent human, mouse and rat TFs. Notable features of the TFe website include a high-quality PDF generator and web API for programmatic data retrieval. TFe aims to rapidly educate scientists about the TFs they encounter through the delivery of succinct summaries written and vetted by experts in the field. TFe is available at http://www.cisreg.ca/tfe.
- Published
- 2012
12. Twist contributes to hormone resistance in breast cancer by downregulating estrogen receptor-α
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Vesuna, F, primary, Lisok, A, additional, Kimble, B, additional, Domek, J, additional, Kato, Y, additional, van der Groep, P, additional, Artemov, D, additional, Kowalski, J, additional, Carraway, H, additional, van Diest, P, additional, and Raman, V, additional
- Published
- 2011
- Full Text
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13. The transcription factor encyclopedia
- Author
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Yusuf, D., Butland, S. L., Swanson, M. I., Bolotin, E., Ticoll, A., Cheung, W. A., Zhang, X. Y., Dickman, C. T., Fulton, D. L., Lim, J. S., Schnabl, J. M., Ramos, O. H., Vasseur-Cognet, M., Leeuw, C. N., Simpson, E. M., Ryffel, G. U., Lam, E. W., Kist, R., Wilson, M. S., Marco-Ferreres, R., Brosens, J. J., Beccari, L. L., Bovolenta, P., Benayoun, B. A., Monteiro, L. J., Schwenen, H. D., Grontved, L., Wederell, E., Mandrup, S., Veitia, R. A., Chakravarthy, H., Hoodless, P. A., Mancarelli, M. M., Torbett, B. E., Banham, A. H., Reddy, S. P., Cullum, R. L., Liedtke, M., Tschan, M. P., Vaz, M., Rizzino, A., Zannini, M., Frietze, S., Farnham, P. J., Eijkelenboom, A., Brown, P. J., Laperrière, D., Leprince, D., Cristofaro, T., Prince, K. L., Putker, M., Del Peso, L., Camenisch, G., Wenger, R. H., Mikula, M., Rozendaal, M., Mader, S., Ostrowski, J., Rhodes, S. J., Rechem, C., Boulay, G., Olechnowicz, S. W., Breslin, M. B., Lan, M. S., Nanan, K. K., Wegner, M., Hou, J., Mullen, R. D., Colvin, S. C., Noy, P. J., Webb, C. F., Witek, M. E., Ferrell, S., Daniel, J. M., Park, J., Waldman, S. A., Peet, D. J., Taggart, M., Jayaraman, P. S., Karrich, J. J., Blom, B., Vesuna, F., O Geen, H., Sun, Y., Gronostajski, R. M., Woodcroft, M. W., Margaret Hough, Chen, E., Europe-Finner, G. N., Karolczak-Bayatti, M., Bailey, J., Hankinson, O., Raman, V., Lebrun, D. P., Biswal, S., Harvey, C. J., Debruyne, J. P., Hogenesch, J. B., and Hevner, R. F.
14. Targeting RNA helicase DDX3X with a small molecule inhibitor for breast cancer bone metastasis treatment.
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Winnard PT Jr, Vesuna F, Bol GM, Gabrielson KL, Chenevix-Trench G, Ter Hoeve ND, van Diest PJ, and Raman V
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- Animals, Humans, Female, Cell Line, Tumor, Mice, Antineoplastic Agents pharmacology, Antineoplastic Agents therapeutic use, Drug Resistance, Neoplasm, Azepines, Imidazoles, DEAD-box RNA Helicases metabolism, DEAD-box RNA Helicases antagonists & inhibitors, DEAD-box RNA Helicases genetics, Bone Neoplasms secondary, Bone Neoplasms drug therapy, Breast Neoplasms drug therapy, Breast Neoplasms pathology, Breast Neoplasms metabolism, Breast Neoplasms genetics
- Abstract
Patients who present with breast cancer bone metastasis only have limited palliative treatment strategies and efficacious drug treatments are needed. In breast cancer patient data, high levels of the RNA helicase DDX3 are associated with poor overall survival and bone metastasis. Consequently, our objective was to target DDX3 in a mouse breast cancer bone metastasis model using a small molecule inhibitor of DDX3, RK-33. Histologically confirmed live imaging indicated no bone metastases in the RK-33 treated cohort, as opposed to placebo-treated mice. We generated a cell line from a bone metastatic lesion in mouse and found that it along with a patient-derived bone metastasis cell line gained resistance to conventional chemotherapeutics but not to RK-33. Finally, differential levels of DDX3 were observed in breast cancer patient metastatic bone samples. Overall, this study indicates that DDX3 is a relevant clinical target in breast cancer bone metastasis and that RK-33 can be a safe and effective treatment for these patients., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)
- Published
- 2024
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15. Twist alters the breast tumor microenvironment via choline kinase to facilitate an aggressive phenotype.
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Vesuna F, Penet MF, Mori N, Bhujwalla ZM, and Raman V
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- Cell Line, Tumor, Choline metabolism, Phenotype, Tumor Microenvironment, Twist-Related Protein 1 genetics, Twist-Related Protein 1 metabolism, Choline Kinase genetics, Choline Kinase metabolism, Phosphorylcholine metabolism
- Abstract
Twist (TWIST1) is a gene required for cell fate specification in embryos and its expression in mammary epithelium can initiate tumorigenesis through the epithelial-mesenchymal transition. To identify downstream target genes of Twist in breast cancer, we performed microarray analysis on the transgenic breast cancer cell line, MCF-7/Twist. One of the targets identified was choline kinase whose upregulation resulted in increased cellular phosphocholine and total choline containing compounds-a characteristic observed in highly aggressive metastatic cancers. To study the interactions between Twist, choline kinase, and their effect on the microenvironment, we used
1 H magnetic resonance spectroscopy and found significantly higher phosphocholine and total choline, as well as increased phosphocholine/glycerophosphocholine ratio in MCF-7/Twist cells. We also observed significant increases in extracellular glucose, lactate, and [H +] ion concentrations in the MCF-7/Twist cells. Magnetic resonance imaging of MCF-7/Twist orthotopic breast tumors showed a significant increase in vascular volume and permeability surface area product compared to control tumors. In addition, by reverse transcription-quantitative polymerase chain reaction, we discovered that Twist upregulated choline kinase expression in estrogen receptor negative breast cancer cell lines through FOXA1 downregulation. Moreover, using The Cancer Genome Atlas database, we observed a significant inverse relationship between FOXA1 and choline kinase expression and propose that it could act as a modulator of the Twist/choline kinase axis. The data presented indicate that Twist is a driver of choline kinase expression in breast cancer cells via FOXA1 resulting in the generation of an aggressive breast cancer phenotype., (© 2022. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)- Published
- 2023
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16. Functional roles of FAP-α in metabolism, migration and invasion of human cancer cells.
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Mori N, Jin J, Krishnamachary B, Mironchik Y, Wildes F, Vesuna F, Barnett JD, and Bhujwalla ZM
- Abstract
Fibroblast activation protein-α (FAP-α) is a transmembrane serine protease that is attracting significant interest as it is expressed by a subgroup of cancer-associated fibroblasts that play a role in immune suppression and cancer metastasis. FAP-α is also expressed by some cancer cells, such as melanoma, colorectal and breast cancer cells. Triple negative breast cancer (TNBC) is an aggressive cancer that urgently requires identification of novel targets for therapy. To expand our understanding of the functional roles of FAP-α in TNBC we engineered a human TNBC cell line, MDA-MB-231, to stably overexpress FAP-α and characterized changes in metabolism by
1 H magnetic resonance spectroscopy, cell proliferation, migration characterized by wound healing, and invasion. FAP-α overexpression resulted in significant alterations in myoinositol, choline metabolites, creatine, and taurine, as well as a significant increase of migration and invasion, although proliferation remained unaltered. The increase of migration and invasion are consistent with the known activities of FAP-α as an exopeptidase and endopeptidase/gelatinase/collagenase in tissue remodeling and repair, and in cell migration. We additionally determined the effects of FAP-α overexpression on the human fibrosarcoma HT1080 cell line that showed increased migration, accompanied by limited changes in metabolism that identified the dependency of the metabolic changes on cell type. These metabolic data identify a previously unknown role of FAP-α in modifying cancer cell metabolism in the TNBC cell line studied here that may provide new insights into its functional roles in cancer progression., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Mori, Jin, Krishnamachary, Mironchik, Wildes, Vesuna, Barnett and Bhujwalla.)- Published
- 2023
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17. RK-33, a small molecule inhibitor of host RNA helicase DDX3, suppresses multiple variants of SARS-CoV-2.
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Vesuna F, Akhrymuk I, Smith A, Winnard PT Jr, Lin SC, Panny L, Scharpf R, Kehn-Hall K, and Raman V
- Abstract
SARS-CoV-2, the virus behind the deadly COVID-19 pandemic, continues to spread globally even as vaccine strategies are proving effective in preventing hospitalizations and deaths. However, evolving variants of the virus appear to be more transmissive and vaccine efficacy toward them is waning. As a result, SARS-CoV-2 will continue to have a deadly impact on public health into the foreseeable future. One strategy to bypass the continuing problem of newer variants is to target host proteins required for viral replication. We have used this host-targeted antiviral (HTA) strategy that targets DDX3X (DDX3), a host DEAD-box RNA helicase that is usurped by SARS-CoV-2 for virus production. We demonstrated that targeting DDX3 with RK-33, a small molecule inhibitor, reduced the viral load in four isolates of SARS-CoV-2 (Lineage A, and Lineage B Alpha, Beta, and Delta variants) by one to three log orders in Calu-3 cells. Furthermore, proteomics and RNA-seq analyses indicated that most SARS-CoV-2 genes were downregulated by RK-33 treatment. Also, we show that the use of RK-33 decreases TMPRSS2 expression, which may be due to DDX3s ability to unwind G-quadraplex structures present in the TMPRSS2 promoter. The data presented support the use of RK-33 as an HTA strategy to control SARS-CoV-2 infection, irrespective of its mutational status, in humans., Competing Interests: VR holds a patent on the composition of RK-33 (US patent # 8518901). The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Vesuna, Akhrymuk, Smith, Winnard, Lin, Panny, Scharpf, Kehn-Hall and Raman.)
- Published
- 2022
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18. Twist activates miR-22 to suppress estrogen receptor alpha in breast cancer.
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Vesuna F, Lisok A, van Diest P, and Raman V
- Subjects
- Breast Neoplasms genetics, Breast Neoplasms pathology, Estrogen Receptor alpha genetics, Female, Humans, MCF-7 Cells, MicroRNAs genetics, Neoplasm Proteins genetics, Nuclear Proteins genetics, RNA, Neoplasm genetics, Twist-Related Protein 1 genetics, Breast Neoplasms metabolism, Estrogen Receptor alpha metabolism, MicroRNAs metabolism, Neoplasm Proteins metabolism, Nuclear Proteins metabolism, RNA, Neoplasm metabolism, Twist-Related Protein 1 metabolism
- Abstract
TWIST1 (Twist) is a basic helix-loop-helix transcription factor that is overexpressed in many cancers and promotes tumor cell invasion, metastasis, and recurrence. In this study, we demonstrate that Twist upregulates expression of microRNA 22 (miR-22) which, in turn, downregulates estrogen receptor alpha (ER) expression in breast cancer. Initial analysis of miR-22 and Twist expression in a panel of breast cancer cell lines showed a direct correlation between Twist and miR-22 levels with miR-22 being highly expressed in ER negative cell lines. Overexpressing Twist caused increased miR-22 levels while downregulating it led to decreased miR-22 expression. To characterize the upstream promoter region of miR-22, we utilized rapid amplification of cDNA ends and identified the transcription start site and the putative promoter region of miR-22. Mechanistically, we determined that Twist, in combination with HDAC1 and DNMT3B, transcriptionally upregulates miR-22 expression by binding to E-boxes in the proximal miR-22 promoter. We also established that miR-22 causes an increase in growth in 3D but not 2D cultures. Importantly, we observed a direct correlation between increased breast cancer grade and Twist and miR-22 expression. We also identified two potential miR-22 binding sites in the 3'-UTR region of ER and confirmed by promoter assays that miR-22 regulates ER expression by binding to both target sites. These results reveal a novel pathway of ER suppression by Twist through miR-22 activation that could potentially promote the ER negative phenotype in breast cancers.
- Published
- 2021
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19. Targeting host DEAD-box RNA helicase DDX3X for treating viral infections.
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Winnard PT Jr, Vesuna F, and Raman V
- Subjects
- DEAD-box RNA Helicases genetics, Humans, Immunity, Innate, Antiviral Agents therapeutic use, DEAD-box RNA Helicases antagonists & inhibitors, Host-Pathogen Interactions drug effects, Virus Diseases drug therapy, Virus Replication drug effects, Viruses drug effects
- Abstract
DDX3X or DDX3, a member of the DEAD (asp, glu, ala, asp) box RNA helicase family of proteins, is a multifunctional protein, which is usurped by several viruses and is vital to their production. To date, 18 species of virus from 12 genera have been demonstrated to be dependent on DDX3 for virulence. In addition, DDX3 has been shown to function within 7 of 10 subcellular regions that are involved in the metabolism of viruses. As such, due to its direct interaction with viral components across most or all stages of viral life cycles, DDX3 can be considered an excellent host target for pan-antiviral drug therapy and has been reported to be a possible broad-spectrum antiviral target. Along these lines, it has been demonstrated that treatment of virally infected cells with small molecule inhibitors of DDX3 blunts virion productions. On the other hand, DDX3 bolsters an innate immune response and viruses have evolved capacities to sequester or block DDX3, which dampens an innate immune response. Thus, enhancing DDX3 production or co-targeting direct viral products that interfere with DDX3's modulation of innate immunity would also diminish virion production. Here we review the evidence that supports the hypothesis that modulating DDX3's agonistic and antagonistic functions during viral infections could have an important impact on safely and efficiently subduing a broad-spectrum of viral infections., (Copyright © 2020 Elsevier B.V. All rights reserved.)
- Published
- 2021
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20. Divergent organ-specific isogenic metastatic cell lines identified using multi-omics exhibit differential drug sensitivity.
- Author
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Winnard PT Jr, Vesuna F, Muthukumar S, and Raman V
- Subjects
- Biomarkers, Pharmacological metabolism, Female, Humans, Neoplasm Metastasis prevention & control, Pharmaceutical Preparations metabolism, Proteomics methods, Breast Neoplasms pathology, Cell Line, Tumor pathology, Neoplasm Metastasis physiopathology
- Abstract
Background: Monitoring and treating metastatic progression remains a formidable task due, in part, to an inability to monitor specific differential molecular adaptations that allow the cancer to thrive within different tissue types. Hence, to develop optimal treatment strategies for metastatic disease, an important consideration is the divergence of the metastatic cancer growing in visceral organs from the primary tumor. We had previously reported the establishment of isogenic human metastatic breast cancer cell lines that are representative of the common metastatic sites observed in breast cancer patients., Methods: Here we have used proteomic, RNAseq, and metabolomic analyses of these isogenic cell lines to systematically identify differences and commonalities in pathway networks and examine the effect on the sensitivity to breast cancer therapeutic agents., Results: Proteomic analyses indicated that dissemination of cells from the primary tumor sites to visceral organs resulted in cell lines that adapted to growth at each new site by, in part, acquiring protein pathways characteristic of the organ of growth. RNAseq and metabolomics analyses further confirmed the divergences, which resulted in differential efficacies to commonly used FDA approved chemotherapeutic drugs. This model system has provided data that indicates that organ-specific growth of malignant lesions is a selective adaptation and growth process., Conclusions: The insights provided by these analyses indicate that the rationale of targeted treatment of metastatic disease may benefit from a consideration that the biology of metastases has diverged from the primary tumor biology and using primary tumor traits as the basis for treatment may not be ideal to design treatment strategies., Competing Interests: The authors have read the journal’s policy and have the following competing interest: VR is the Founder and Director of Natsar Pharmaceuticals Inc. The authors would like to declare the following patents associated with this research: VR holds a patent on the composition of RK-33 (US patent # 8,518,901). This does not alter our adherence to PLOS ONE policies on sharing data and materials.
- Published
- 2020
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21. Targeting RNA helicase DDX3 in stem cell maintenance and teratoma formation.
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Kerr CL, Bol GM, Vesuna F, and Raman V
- Abstract
DDX3 is an RNA helicase that has antiapoptotic properties, and promotes proliferation and transformation. Besides the role of DDX3 in transformed cells, there is evidence to indicate that DDX3 expression is at its highest levels during early embryonic development and is also expressed in germ cells of adults. Even though there is a distinct pattern of DDX3 expression during embryonic development and in adults, very little is known regarding its role in embryonic stem cells and pluripotency. In this work, we examined the relationship between DDX3 and human embryonic stem cells and its differentiated lineages. DDX3 expression was analyzed by immunohistochemistry in human embryonic stem cells and embryonal carcinoma cells. From the data obtained, it was evident that DDX3 was overexpressed in undifferentiated stem cells compared to differentiated cells. Moreover, when DDX3 expression was abrogated in multiple stem cells, proliferation was decreased, but differentiation was facilitated. Importantly, this resulted in reduced potency to induce teratoma formation. Taken together, these findings indicate a distinct role for DDX3 in stem cell maintenance., Competing Interests: CONFLICT OF INTEREST Venu Raman has a patent for RK-33 and is on the advisory board of Natsar Pharmaceuticals.
- Published
- 2019
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22. Targeting DDX3 in Medulloblastoma Using the Small Molecule Inhibitor RK-33.
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Tantravedi S, Vesuna F, Winnard PT Jr, Martin A, Lim M, Eberhart CG, Berlinicke C, Raabe E, van Diest PJ, and Raman V
- Abstract
Medulloblastoma is the most common malignant tumor that arises from the cerebellum of the central nervous system. Clinically, medulloblastomas are treated by surgery, radiation, and chemotherapy, all of which result in toxicity and morbidity. Recent reports have identified that DDX3, a member of the RNA helicase family, is mutated in medulloblastoma. In this study, we demonstrate the role of DDX3 in driving medulloblastoma. With the use of a small molecule inhibitor of DDX3, RK-33, we could inhibit growth and promote cell death in two medulloblastoma cell lines, DAOY and UW228, with IC50 values of 2.5 μM and 3.5 μM, respectively. Treatment of DAOY and UW228 cells with RK-33 caused a G1 arrest, resulted in reduced TCF reporter activity, and reduced mRNA expression levels of downstream target genes of the WNT pathway, such as Axin2, CCND1, MYC, and Survivin. In addition, treatment of DAOY and UW228 cells with a combination of RK-33 and radiation exhibited a synergistic effect. Importantly, the combination of RK-33 and 5 Gy radiation caused tumor regression in a mouse xenograft model of medulloblastoma. Using immunohistochemistry, we observed DDX3 expression in both pediatric (55%) and adult (66%) medulloblastoma patients. Based on these results, we conclude that RK-33 is a promising radiosensitizing agent that inhibits DDX3 activity and down-regulates WNT/β-catenin signaling and could be used as a frontline therapeutic strategy for DDX3-expressing medulloblastomas in combination with radiation., (Copyright © 2018 The Authors. Published by Elsevier Inc. All rights reserved.)
- Published
- 2019
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23. Global Effects of DDX3 Inhibition on Cell Cycle Regulation Identified by a Combined Phosphoproteomics and Single Cell Tracking Approach.
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Heerma van Voss MR, Kammers K, Vesuna F, Brilliant J, Bergman Y, Tantravedi S, Wu X, Cole RN, Holland A, van Diest PJ, and Raman V
- Abstract
DDX3 is an RNA helicase with oncogenic properties. The small molecule inhibitor RK-33 is designed to fit into the ATP binding cleft of DDX3 and hereby block its activity. RK-33 has shown potent activity in preclinical cancer models. However, the mechanism behind the antineoplastic activity of RK-33 remains largely unknown. In this study we used a dual phosphoproteomic and single cell tracking approach to evaluate the effect of RK-33 on cancer cells. MDA-MB-435 cells were treated for 24 hours with RK-33 or vehicle control. Changes in phosphopeptide abundance were analyzed with quantitative mass spectrometry using isobaric mass tags (Tandem Mass Tags). At the proteome level we mainly observed changes in mitochondrial translation, cell division pathways and proteins related to cell cycle progression. Analysis of the phosphoproteome indicated decreased CDK1 activity after RK-33 treatment. To further evaluate the effect of DDX3 inhibition on cell cycle progression over time, we performed timelapse microscopy of Fluorescent Ubiquitin Cell Cycle Indicators labeled cells after RK-33 or siDDX3 exposure. Single cell tracking indicated that DDX3 inhibition resulted in a global delay in cell cycle progression in interphase and mitosis. In addition, we observed an increase in endoreduplication. Overall, we conclude that DDX3 inhibition affects cells in all phases and causes a global cell cycle progression delay., (Copyright © 2018 The Authors. Published by Elsevier Inc. All rights reserved.)
- Published
- 2018
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24. Role of DDX3 in the pathogenesis of inflammatory bowel disease.
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Tantravedi S, Vesuna F, Winnard PT Jr, Van Voss MRH, Van Diest PJ, and Raman V
- Abstract
When crypt stem cells of the gastrointestinal tract become injured, the result is increased synthesis of pro-inflammatory cytokines and matrix metalloproteinases by their progeny - the colonic epithelium. Chronic inflammation of the gastrointestinal tract is a characteristic of inflammatory bowel disease, which includes Crohn's Disease and Ulcerative Colitis. In our ongoing investigation to decipher the characteristic functions of a RNA helicase gene, DDX3, we identified high DDX3 expression by immunohistochemistry of colon biopsy samples, which included chronic/mild Morbus Crohn, active Morbus Crohn, Chronic/mild Colitis Ulcerosa and active Colitis Ulcerosa in epithelium and stromal compartments. We used a small molecule inhibitor of DDX3, RK-33, on two human colonic epithelial cell lines, HCEC1CT and HCEC2CT and found that RK-33 was able to decrease expression of MMP-1, MMP-2, MMP-3, and MMP-10. Moreover, forced differentiation of a human colonic cancer cell line, HT29, resulted in decreased DDX3 levels, indicating that DDX3 contributes to the modulation of colonic epithelium differentiation. In conclusion, our results revealed novel functions of DDX3 in inflammatory bowel disease and indicate a potential for using RK-33 as a systemic therapy to promote not only differentiation of transformed colonic epithelium but also to reduce MMP expression and thus elicit a decreased inflammatory response., Competing Interests: CONFLICTS OF INTEREST The authors report no conflicts of interest.
- Published
- 2017
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25. Organ-specific isogenic metastatic breast cancer cell lines exhibit distinct Raman spectral signatures and metabolomes.
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Winnard PT Jr, Zhang C, Vesuna F, Kang JW, Garry J, Dasari RR, Barman I, and Raman V
- Subjects
- Animals, Breast Neoplasms pathology, Cell Line, Tumor, Female, Heterografts, Humans, Mice, Mice, Inbred NOD, Mice, SCID, Neoplasm Metastasis pathology, Spectrum Analysis, Raman, Breast Neoplasms chemistry, Breast Neoplasms metabolism, Metabolome
- Abstract
Molecular characterization of organ-specific metastatic lesions, which distinguish them from the primary tumor, will provide a better understanding of tissue specific adaptations that regulate metastatic progression. Using an orthotopic xenograft model, we have isolated isogenic metastatic human breast cancer cell lines directly from organ explants that are phenotypically distinct from the primary tumor cell line. Label-free Raman spectroscopy was used and informative spectral bands were ascertained as differentiators of organ-specific metastases as opposed to the presence of a single universal marker. Decision algorithms derived from the Raman spectra unambiguously identified these isogenic cell lines as unique biological entities - a finding reinforced through metabolomic analyses that indicated tissue of origin metabolite distinctions between the cell lines. Notably, complementarity of the metabolomics and Raman datasets was found. Our findings provide evidence that metastatic spread generates tissue-specific adaptations at the molecular level within cancer cells, which can be differentiated with Raman spectroscopy.
- Published
- 2017
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26. Combination treatment using DDX3 and PARP inhibitors induces synthetic lethality in BRCA1-proficient breast cancer.
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Heerma van Voss MR, Brilliant JD, Vesuna F, Bol GM, van der Wall E, van Diest PJ, and Raman V
- Subjects
- Adult, Azepines administration & dosage, Azepines pharmacology, BRCA1 Protein genetics, Breast Neoplasms enzymology, Breast Neoplasms genetics, Cell Line, Tumor, DEAD-box RNA Helicases biosynthesis, DEAD-box RNA Helicases metabolism, Drug Synergism, Enzyme Inhibitors administration & dosage, Enzyme Inhibitors pharmacology, Female, Genes, BRCA1, Germ-Line Mutation, Humans, Imidazoles administration & dosage, Imidazoles pharmacology, MCF-7 Cells, Middle Aged, Phthalazines administration & dosage, Phthalazines pharmacology, Piperazines administration & dosage, Piperazines pharmacology, Poly(ADP-ribose) Polymerase Inhibitors administration & dosage, Poly(ADP-ribose) Polymerase Inhibitors pharmacology, Poly(ADP-ribose) Polymerases biosynthesis, Poly(ADP-ribose) Polymerases metabolism, Antineoplastic Combined Chemotherapy Protocols pharmacology, Breast Neoplasms drug therapy, DEAD-box RNA Helicases antagonists & inhibitors
- Abstract
Triple-negative breast cancers have unfavorable outcomes due to their inherent aggressive behavior and lack of targeted therapies. Breast cancers occurring in BRCA1 mutation carriers are mostly triple-negative and harbor homologous recombination deficiency, sensitizing them to inhibition of a second DNA damage repair pathway by, e.g., PARP inhibitors. Unfortunately, resistance against PARP inhibitors in BRCA1-deficient cancers is common and sensitivity is limited in BRCA1-proficient breast cancers. RK-33, an inhibitor of the RNA helicase DDX3, was previously demonstrated to impede non-homologous end-joining repair of DNA breaks. Consequently, we evaluated DDX3 as a therapeutic target in BRCA pro- and deficient breast cancers and assessed whether DDX3 inhibition could sensitize cells to PARP inhibition. High DDX3 expression was identified by immunohistochemistry in breast cancer samples of 24% of BRCA1 (p = 0.337) and 21% of BRCA2 mutation carriers (p = 0.624), as compared to 30% of sporadic breast cancer samples. The sensitivity to the DDX3 inhibitor RK-33 was similar in BRCA1 pro- and deficient breast cancer cell lines, with IC50 values in the low micromolar range (2.8-6.6 μM). A synergistic interaction was observed for combination treatment with RK-33 and the PARP inhibitor olaparib in BRCA1-proficient breast cancer, with the mean combination index ranging from 0.59 to 0.62. Overall, we conclude that BRCA pro- and deficient breast cancers have a similar dependency upon DDX3. DDX3 inhibition by RK-33 synergizes with PARP inhibitor treatment, especially in breast cancers with a BRCA1-proficient background.
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- 2017
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27. Genomic pathways modulated by Twist in breast cancer.
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Vesuna F, Bergman Y, and Raman V
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- Animals, Cell Line, Tumor, Cell Proliferation genetics, Female, Gene Expression Profiling methods, Gene Expression Regulation, Neoplastic genetics, Genomics methods, Humans, Lung Neoplasms genetics, MCF-7 Cells, Mammary Neoplasms, Animal genetics, Mice, Mice, SCID, Breast Neoplasms genetics, Signal Transduction genetics, Twist-Related Protein 1 genetics
- Abstract
Background: The basic helix-loop-helix transcription factor TWIST1 (Twist) is involved in embryonic cell lineage determination and mesodermal differentiation. There is evidence to indicate that Twist expression plays a role in breast tumor formation and metastasis, but the role of Twist in dysregulating pathways that drive the metastatic cascade is unclear. Moreover, many of the genes and pathways dysregulated by Twist in cell lines and mouse models have not been validated against data obtained from larger, independant datasets of breast cancer patients., Methods: We over-expressed the human Twist gene in non-metastatic MCF-7 breast cancer cells to generate the estrogen-independent metastatic breast cancer cell line MCF-7/Twist. These cells were inoculated in the mammary fat pad of female severe compromised immunodeficient mice, which subsequently formed xenograft tumors that metastasized to the lungs. Microarray data was collected from both in vitro (MCF-7 and MCF-7/Twist cell lines) and in vivo (primary tumors and lung metastases) models of Twist expression. Our data was compared to several gene datasets of various subtypes, classes, and grades of human breast cancers., Results: Our data establishes a Twist over-expressing mouse model of breast cancer, which metastasizes to the lung and replicates some of the ontogeny of human breast cancer progression. Gene profiling data, following Twist expression, exhibited novel metastasis driver genes as well as cellular maintenance genes that were synonymous with the metastatic process. We demonstrated that the genes and pathways altered in the transgenic cell line and metastatic animal models parallel many of the dysregulated gene pathways observed in human breast cancers., Conclusions: Analogous gene expression patterns were observed in both in vitro and in vivo Twist preclinical models of breast cancer metastasis and breast cancer patient datasets supporting the functional role of Twist in promoting breast cancer metastasis. The data suggests that genetic dysregulation of Twist at the cellular level drives alterations in gene pathways in the Twist metastatic mouse model which are comparable to changes seen in human breast cancers. Lastly, we have identified novel genes and pathways that could be further investigated as targets for drugs to treat metastatic breast cancer.
- Published
- 2017
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28. RK-33 Radiosensitizes Prostate Cancer Cells by Blocking the RNA Helicase DDX3.
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Xie M, Vesuna F, Tantravedi S, Bol GM, Heerma van Voss MR, Nugent K, Malek R, Gabrielson K, van Diest PJ, Tran PT, and Raman V
- Subjects
- Animals, Cell Cycle Checkpoints drug effects, Cell Line, Tumor, Cell Movement drug effects, Cell Proliferation drug effects, DEAD-box RNA Helicases analysis, DEAD-box RNA Helicases physiology, DNA Damage, Histones analysis, Humans, Male, Mice, Prostatic Neoplasms chemistry, Prostatic Neoplasms pathology, Azepines pharmacology, DEAD-box RNA Helicases antagonists & inhibitors, Imidazoles pharmacology, Prostatic Neoplasms radiotherapy, Radiation-Sensitizing Agents pharmacology
- Abstract
Despite advances in diagnosis and treatment, prostate cancer is the most prevalent cancer in males and the second highest cause of cancer-related mortality. We identified an RNA helicase gene, DDX3 (DDX3X), which is overexpressed in prostate cancers, and whose expression is directly correlated with high Gleason scores. Knockdown of DDX3 in the aggressive prostate cancer cell lines DU145 and 22Rv1 resulted in significantly reduced clonogenicity. To target DDX3, we rationally designed a small molecule, RK-33, which docks into the ATP-binding domain of DDX3. Functional studies indicated that RK-33 preferentially bound to DDX3 and perturbed its activity. RK-33 treatment of prostate cancer cell lines DU145, 22Rv1, and LNCaP (which have high DDX3 levels) decreased proliferation and induced a G
1 phase cell-cycle arrest. Conversely, the low DDX3-expressing cell line, PC3, exhibited few changes following RK-33 treatment. Importantly, combination studies using RK-33 and radiation exhibited synergistic effects both in vitro and in a xenograft model of prostate cancer demonstrating the role of RK-33 as a radiosensitizer. Taken together, these results indicate that blocking DDX3 by RK-33 in combination with radiation treatment is a viable option for treating locally advanced prostate cancer. Cancer Res; 76(21); 6340-50. ©2016 AACR., (©2016 American Association for Cancer Research.)- Published
- 2016
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29. miRNA expression patterns in normal breast tissue and invasive breast cancers of BRCA1 and BRCA2 germ-line mutation carriers.
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Vos S, Vesuna F, Raman V, van Diest PJ, and van der Groep P
- Subjects
- Adult, Aged, Aged, 80 and over, Biomarkers, Tumor genetics, Breast Neoplasms metabolism, Breast Neoplasms pathology, Carcinoma, Ductal, Breast genetics, Carcinoma, Ductal, Breast metabolism, Carcinoma, Ductal, Breast pathology, Carcinoma, Lobular genetics, Carcinoma, Lobular metabolism, Carcinoma, Lobular pathology, Carcinoma, Medullary genetics, Carcinoma, Medullary metabolism, Carcinoma, Medullary pathology, Case-Control Studies, Chromosomes, Human genetics, Female, Follow-Up Studies, Genetic Predisposition to Disease, Humans, Immunoenzyme Techniques, Microarray Analysis, Middle Aged, Neoplasm Grading, Neoplasm Invasiveness, Prognosis, RNA, Messenger genetics, Real-Time Polymerase Chain Reaction, Receptor, ErbB-2 metabolism, Receptors, Estrogen metabolism, Receptors, Progesterone metabolism, Reverse Transcriptase Polymerase Chain Reaction, Young Adult, BRCA1 Protein genetics, BRCA2 Protein genetics, Breast Neoplasms genetics, Germ-Line Mutation genetics, MicroRNAs genetics
- Abstract
miRNA deregulation has been found to promote carcinogenesis. Little is known about miRNA deregulation in hereditary breast tumors as no miRNA expression profiling studies have been performed in normal breast tissue of BRCA1 and BRCA2 mutation carriers. miRNA profiles of 17 BRCA1- and 9 BRCA2-associated breast carcinomas were analyzed using microarrays. Normal breast tissues from BRCA1 and BRCA2 mutation carriers (both n = 5) and non-mutation carriers (n = 10) were also included. Candidate miRNAs were validated by qRT-PCR. Breast carcinomas showed extensive miRNA alteration compared to normal breast tissues in BRCA1 and BRCA2 mutation carriers. Moreover, normal breast tissue from BRCA1 mutation carriers already showed miRNA alterations compared to non-mutation carriers. Chromosomal distribution analysis showed several hotspots containing down- or up-regulated miRNAs. Pathway analysis yielded many similarities between the BRCA1 and BRCA2 axes with miRNAs involved in cell cycle regulation, proliferation and apoptosis. Lesser known pathways were also affected, including cellular movement and protein trafficking. This study provides a comprehensive insight into the potential role of miRNA deregulation in BRCA1/2-associated breast carcinogenesis. The observed extensive miRNA deregulation is likely the result of genome-wide effects of chromosomal instability caused by impaired BRCA1 or BRCA2 function. This study's results also suggest the existence of common pathways driving breast carcinogenesis in both BRCA1 and BRCA2 germ-line mutation carriers.
- Published
- 2015
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30. NZ51, a ring-expanded nucleoside analog, inhibits motility and viability of breast cancer cells by targeting the RNA helicase DDX3.
- Author
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Xie M, Vesuna F, Botlagunta M, Bol GM, Irving A, Bergman Y, Hosmane RS, Kato Y, Winnard PT Jr, and Raman V
- Subjects
- Animals, Antineoplastic Agents chemistry, Antineoplastic Agents pharmacology, Azepines chemistry, Breast Neoplasms genetics, Breast Neoplasms metabolism, Cell Line, Cell Line, Tumor, Cell Movement genetics, Cell Proliferation drug effects, Cell Proliferation genetics, Cell Survival drug effects, Cell Survival genetics, DEAD-box RNA Helicases genetics, DEAD-box RNA Helicases metabolism, Female, G1 Phase Cell Cycle Checkpoints drug effects, G1 Phase Cell Cycle Checkpoints genetics, Humans, Immunoblotting, MCF-7 Cells, Mice, Nude, Molecular Structure, Nucleosides chemistry, RNA Interference, Tumor Burden drug effects, Tumor Burden genetics, Xenograft Model Antitumor Assays, Azepines pharmacology, Breast Neoplasms drug therapy, Cell Movement drug effects, DEAD-box RNA Helicases antagonists & inhibitors, Nucleosides pharmacology
- Abstract
DDX3X (DDX3), a human RNA helicase, is over expressed in multiple breast cancer cell lines and its expression levels are directly correlated to cellular aggressiveness. NZ51, a ring-expanded nucleoside analogue (REN) has been reported to inhibit the ATP dependent helicase activity of DDX3. Molecular modeling of NZ51 binding to DDX3 indicated that the 5:7-fused imidazodiazepine ring of NZ51 was incorporated into the ATP binding pocket of DDX3. In this study, we investigated the anticancer properties of NZ51 in MCF-7 and MDA-MB-231 breast cancer cell lines. NZ51 treatment decreased cellular motility and cell viability of MCF-7 and MDA-MB-231 cells with IC50 values in the low micromolar range. Biological knockdown of DDX3 in MCF-7 and MDA-MB-231 cells resulted in decreased proliferation rates and reduced clonogenicity. In addition, NZ51 was effective in killing breast cancer cells under hypoxic conditions with the same potency as observed during normoxia. Mechanistic studies indicated that NZ51 did not cause DDX3 degradation, but greatly diminished its functionality. Moreover, in vivo experiments demonstrated that DDX3 knockdown by shRNA resulted in reduced tumor volume and metastasis without altering tumor vascular volume or permeability-surface area. In initial in vivo experiments, NZ51 treatment did not significantly reduce tumor volume. Further studies are needed to optimize drug formulation, dose and delivery. Continuing work will determine the in vitro-in vivo correlation of NZ51 activity and its utility in a clinical setting.
- Published
- 2015
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31. Identification of the DEAD box RNA helicase DDX3 as a therapeutic target in colorectal cancer.
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Heerma van Voss MR, Vesuna F, Trumpi K, Brilliant J, Berlinicke C, de Leng W, Kranenburg O, Offerhaus GJ, Bürger H, van der Wall E, van Diest PJ, and Raman V
- Subjects
- Aged, Aged, 80 and over, Azepines pharmacology, Basic Helix-Loop-Helix Leucine Zipper Transcription Factors genetics, Basic Helix-Loop-Helix Leucine Zipper Transcription Factors metabolism, Cell Line, Tumor, Cell Survival drug effects, Cell Survival genetics, Colorectal Neoplasms drug therapy, Colorectal Neoplasms metabolism, DEAD-box RNA Helicases antagonists & inhibitors, DEAD-box RNA Helicases metabolism, Dose-Response Relationship, Drug, Female, Gene Expression Regulation, Neoplastic, HCT116 Cells, HT29 Cells, Humans, Imidazoles pharmacology, Immunoblotting, Immunohistochemistry, Male, Middle Aged, Molecular Targeted Therapy methods, Mutation, RNA Interference, Reverse Transcriptase Polymerase Chain Reaction, Transcription Factor 4, Transcription Factors genetics, Transcription Factors metabolism, Wnt Signaling Pathway drug effects, beta Catenin metabolism, Colorectal Neoplasms genetics, DEAD-box RNA Helicases genetics, Wnt Signaling Pathway genetics, beta Catenin genetics
- Abstract
Identifying druggable targets in the Wnt-signaling pathway can optimize colorectal cancer treatment. Recent studies have identified a member of the RNA helicase family DDX3 (DDX3X) as a multilevel activator of Wnt signaling in cells without activating mutations in the Wnt-signaling pathway. In this study, we evaluated whether DDX3 plays a role in the constitutively active Wnt pathway that drives colorectal cancer. We determined DDX3 expression levels in 303 colorectal cancers by immunohistochemistry. 39% of tumors overexpressed DDX3. High cytoplasmic DDX3 expression correlated with nuclear β-catenin expression, a marker of activated Wnt signaling. Functionally, we validated this finding in vitro and found that inhibition of DDX3 with siRNA resulted in reduced TCF4-reporter activity and lowered the mRNA expression levels of downstream TCF4-regulated genes. In addition, DDX3 knockdown in colorectal cancer cell lines reduced proliferation and caused a G1 arrest, supporting a potential oncogenic role of DDX3 in colorectal cancer. RK-33 is a small molecule inhibitor designed to bind to the ATP-binding site of DDX3. Treatment of colorectal cancer cell lines and patient-derived 3D cultures with RK-33 inhibited growth and promoted cell death with IC50 values ranging from 2.5 to 8 μM. The highest RK-33 sensitivity was observed in tumors with wild-type APC-status and a mutation in CTNNB1. Based on these results, we conclude that DDX3 has an oncogenic role in colorectal cancer. Inhibition of DDX3 with the small molecule inhibitor RK-33 causes inhibition of Wnt signaling and may therefore be a promising future treatment strategy for a subset of colorectal cancers.
- Published
- 2015
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32. Targeting DDX3 with a small molecule inhibitor for lung cancer therapy.
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Bol GM, Vesuna F, Xie M, Zeng J, Aziz K, Gandhi N, Levine A, Irving A, Korz D, Tantravedi S, Heerma van Voss MR, Gabrielson K, Bordt EA, Polster BM, Cope L, van der Groep P, Kondaskar A, Rudek MA, Hosmane RS, van der Wall E, van Diest PJ, Tran PT, and Raman V
- Subjects
- Animals, Antineoplastic Agents isolation & purification, Apoptosis, Azepines isolation & purification, Cell Cycle drug effects, Cell Cycle Checkpoints, Cell Line, Humans, Imidazoles isolation & purification, Mice, Nude, Mice, Transgenic, Radiation-Sensitizing Agents isolation & purification, Antineoplastic Agents pharmacology, Azepines pharmacology, DEAD-box RNA Helicases antagonists & inhibitors, Imidazoles pharmacology, Lung Neoplasms drug therapy, Lung Neoplasms radiotherapy, Radiation-Sensitizing Agents pharmacology
- Abstract
Lung cancer is the most common malignancy worldwide and is a focus for developing targeted therapies due to its refractory nature to current treatment. We identified a RNA helicase, DDX3, which is overexpressed in many cancer types including lung cancer and is associated with lower survival in lung cancer patients. We designed a first-in-class small molecule inhibitor, RK-33, which binds to DDX3 and abrogates its activity. Inhibition of DDX3 by RK-33 caused G1 cell cycle arrest, induced apoptosis, and promoted radiation sensitization in DDX3-overexpressing cells. Importantly, RK-33 in combination with radiation induced tumor regression in multiple mouse models of lung cancer. Mechanistically, loss of DDX3 function either by shRNA or by RK-33 impaired Wnt signaling through disruption of the DDX3-β-catenin axis and inhibited non-homologous end joining-the major DNA repair pathway in mammalian somatic cells. Overall, inhibition of DDX3 by RK-33 promotes tumor regression, thus providing a compelling argument to develop DDX3 inhibitors for lung cancer therapy., (© 2015 The Authors. Published under the terms of the CC BY 4.0 license.)
- Published
- 2015
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33. The twist box domain is required for Twist1-induced prostate cancer metastasis.
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Gajula RP, Chettiar ST, Williams RD, Thiyagarajan S, Kato Y, Aziz K, Wang R, Gandhi N, Wild AT, Vesuna F, Ma J, Salih T, Cades J, Fertig E, Biswal S, Burns TF, Chung CH, Rudin CM, Herman JM, Hales RK, Raman V, An SS, and Tran PT
- Subjects
- Amino Acid Substitution, Animals, Biomarkers, Tumor, Cell Line, Tumor, Epithelial-Mesenchymal Transition genetics, Gene Expression Profiling, Gene Expression Regulation, Neoplastic, Homeodomain Proteins genetics, Homeodomain Proteins metabolism, Humans, Male, Mice, Nude, Neoplasm Invasiveness, Neoplasm Metastasis pathology, Nuclear Proteins genetics, Promoter Regions, Genetic, Prostatic Neoplasms pathology, Transcriptional Activation, Twist-Related Protein 1 genetics, Neoplasm Metastasis genetics, Nuclear Proteins metabolism, Prostatic Neoplasms genetics, Protein Structure, Tertiary genetics, Twist-Related Protein 1 metabolism
- Abstract
Unlabelled: Twist1, a basic helix-loop-helix transcription factor, plays a key role during development and is a master regulator of the epithelial-mesenchymal transition (EMT) that promotes cancer metastasis. Structure-function relationships of Twist1 to cancer-related phenotypes are underappreciated, so we studied the requirement of the conserved Twist box domain for metastatic phenotypes in prostate cancer. Evidence suggests that Twist1 is overexpressed in clinical specimens and correlated with aggressive/metastatic disease. Therefore, we examined a transactivation mutant, Twist1-F191G, in prostate cancer cells using in vitro assays, which mimic various stages of metastasis. Twist1 overexpression led to elevated cytoskeletal stiffness and cell traction forces at the migratory edge of cells based on biophysical single-cell measurements. Twist1 conferred additional cellular properties associated with cancer cell metastasis including increased migration, invasion, anoikis resistance, and anchorage-independent growth. The Twist box mutant was defective for these Twist1 phenotypes in vitro. Importantly, we observed a high frequency of Twist1-induced metastatic lung tumors and extrathoracic metastases in vivo using the experimental lung metastasis assay. The Twist box was required for prostate cancer cells to colonize metastatic lung lesions and extrathoracic metastases. Comparative genomic profiling revealed transcriptional programs directed by the Twist box that were associated with cancer progression, such as Hoxa9. Mechanistically, Twist1 bound to the Hoxa9 promoter and positively regulated Hoxa9 expression in prostate cancer cells. Finally, Hoxa9 was important for Twist1-induced cellular phenotypes associated with metastasis. These data suggest that the Twist box domain is required for Twist1 transcriptional programs and prostate cancer metastasis., Implications: Targeting the Twist box domain of Twist1 may effectively limit prostate cancer metastatic potential., (©2013 AACR.)
- Published
- 2013
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34. Glycerophosphodiester phosphodiesterase domain containing 5 (GDPD5) expression correlates with malignant choline phospholipid metabolite profiles in human breast cancer.
- Author
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Cao MD, Döpkens M, Krishnamachary B, Vesuna F, Gadiya MM, Lønning PE, Bhujwalla ZM, Gribbestad IS, and Glunde K
- Subjects
- Adult, Breast Neoplasms enzymology, Breast Neoplasms genetics, Breast Neoplasms pathology, Cell Extracts, Cell Line, Tumor, Choline Kinase metabolism, Female, Gene Expression Regulation, Neoplastic, Humans, Middle Aged, Nuclear Magnetic Resonance, Biomolecular, Phospholipase D metabolism, Phosphoric Diester Hydrolases genetics, RNA, Messenger genetics, RNA, Messenger metabolism, Breast Neoplasms metabolism, Choline metabolism, Metabolome, Phospholipids metabolism, Phosphoric Diester Hydrolases metabolism
- Abstract
Altered choline phospholipid metabolism is a hallmark of cancer, leading to malignant choline metabolite profiles consisting of low glycerophosphocholine (GPC) and high phosphocholine (PC) in human breast cancers. Glycerophosphocholine phosphodiesterase (GPC-PDE) catalyzes the degradation of GPC to free choline and glycerol-3-phosphate. The gene(s) encoding for the GPC-PDE(s) responsible for GPC degradation in breast cancers have not yet been identified. Here, we demonstrate for the first time that the GPC-PDE encoded by glycerophosphodiester phosphodiesterase domain containing 5 (GDPD5) is associated with breast cancer malignancy. Two human breast cancer cell lines (n = 8 and n = 10) and primary human breast tumor samples (n = 19) were studied with combined MRS and quantitative reverse transcription-polymerase chain reaction to investigate several isoforms of GDPD expression with respect to choline phospholipid metabolite levels. Of the five GDPDs tested, GDPD5 was found to be significantly overexpressed in highly malignant estrogen receptor negative (ER(-)) compared with weakly malignant estrogen receptor positive (ER(+)) human breast cancer cells (p = 0.027) and breast tumors from patients (p = 0.015). GDPD5 showed significantly positive correlations with PC (p < 0.001), total choline (tCho) (p = 0.007) and PC/GPC (p < 0.001) levels in human breast tumors. GDPD5 showed a trend towards a negative correlation with GPC levels (p = 0.130). Human breast cancers with malignant choline metabolite profiles consisting of low GPC and high PC levels highly co-expressed GDPD5, choline kinase alpha (CHKA) and phosphatidylcholine-specific phospholipase D1 (PLD1), whereas cancers containing high GPC and relatively low PC levels displayed low co-expression of GDPD5, CHKA and PLD1. GDPD5, CHKA and PLD1 were significantly overexpressed in highly malignant ER(-) tumors in our patient cohort. Our study identified GDPD5 as a GPC-PDE that probably participates in the regulation of choline phospholipid metabolism in breast cancer, which possibly occurs in cooperation with CHKA and PLD1., (Copyright © 2012 John Wiley & Sons, Ltd.)
- Published
- 2012
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35. The transcription factor encyclopedia.
- Author
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Yusuf D, Butland SL, Swanson MI, Bolotin E, Ticoll A, Cheung WA, Zhang XY, Dickman CT, Fulton DL, Lim JS, Schnabl JM, Ramos OH, Vasseur-Cognet M, de Leeuw CN, Simpson EM, Ryffel GU, Lam EW, Kist R, Wilson MS, Marco-Ferreres R, Brosens JJ, Beccari LL, Bovolenta P, Benayoun BA, Monteiro LJ, Schwenen HD, Grontved L, Wederell E, Mandrup S, Veitia RA, Chakravarthy H, Hoodless PA, Mancarelli MM, Torbett BE, Banham AH, Reddy SP, Cullum RL, Liedtke M, Tschan MP, Vaz M, Rizzino A, Zannini M, Frietze S, Farnham PJ, Eijkelenboom A, Brown PJ, Laperrière D, Leprince D, de Cristofaro T, Prince KL, Putker M, del Peso L, Camenisch G, Wenger RH, Mikula M, Rozendaal M, Mader S, Ostrowski J, Rhodes SJ, Van Rechem C, Boulay G, Olechnowicz SW, Breslin MB, Lan MS, Nanan KK, Wegner M, Hou J, Mullen RD, Colvin SC, Noy PJ, Webb CF, Witek ME, Ferrell S, Daniel JM, Park J, Waldman SA, Peet DJ, Taggart M, Jayaraman PS, Karrich JJ, Blom B, Vesuna F, O'Geen H, Sun Y, Gronostajski RM, Woodcroft MW, Hough MR, Chen E, Europe-Finner GN, Karolczak-Bayatti M, Bailey J, Hankinson O, Raman V, LeBrun DP, Biswal S, Harvey CJ, DeBruyne JP, Hogenesch JB, Hevner RF, Héligon C, Luo XM, Blank MC, Millen KJ, Sharlin DS, Forrest D, Dahlman-Wright K, Zhao C, Mishima Y, Sinha S, Chakrabarti R, Portales-Casamar E, Sladek FM, Bradley PH, and Wasserman WW
- Subjects
- Access to Information, Animals, Encyclopedias as Topic, Humans, Internet, Mice, Rats, Transcription, Genetic, Computational Biology, Databases, Protein supply & distribution, Transcription Factors genetics
- Abstract
Here we present the Transcription Factor Encyclopedia (TFe), a new web-based compendium of mini review articles on transcription factors (TFs) that is founded on the principles of open access and collaboration. Our consortium of over 100 researchers has collectively contributed over 130 mini review articles on pertinent human, mouse and rat TFs. Notable features of the TFe website include a high-quality PDF generator and web API for programmatic data retrieval. TFe aims to rapidly educate scientists about the TFs they encounter through the delivery of succinct summaries written and vetted by experts in the field. TFe is available at http://www.cisreg.ca/tfe.
- Published
- 2012
- Full Text
- View/download PDF
36. Expression of DDX3 is directly modulated by hypoxia inducible factor-1 alpha in breast epithelial cells.
- Author
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Botlagunta M, Krishnamachary B, Vesuna F, Winnard PT Jr, Bol GM, Patel AH, and Raman V
- Subjects
- Animals, Base Sequence, Cell Hypoxia drug effects, Cell Line, Tumor, Chromatin Immunoprecipitation, Cobalt pharmacology, DEAD-box RNA Helicases genetics, Epithelial Cells drug effects, Female, Gene Expression Regulation, Neoplastic drug effects, Humans, Mice, Molecular Sequence Data, Protein Binding drug effects, Protein Stability drug effects, Protein Transport drug effects, Response Elements genetics, Xenograft Model Antitumor Assays, Breast pathology, DEAD-box RNA Helicases metabolism, Epithelial Cells metabolism, Hypoxia-Inducible Factor 1, alpha Subunit metabolism
- Abstract
DEAD box protein, DDX3, is aberrantly expressed in breast cancer cells ranging from weakly invasive to aggressive phenotypes and functions as an important regulator of cancer cell growth and survival. Here, we demonstrate that hypoxia inducible factor-1α is a transcriptional activator of DDX3 in breast cancer cells. Within the promoter region of the human DDX3 gene, we identified three putative hypoxia inducible factor-1 responsive elements. By luciferase reporter assays in combination with mutated hypoxia inducible factor-1 responsive elements, we determined that the hypoxia inducible factor-1 responsive element at position -153 relative to the translation start site is essential for transcriptional activation of DDX3 under hypoxic conditions. We also demonstrated that hypoxia inducible factor-1 binds to the DDX3 promoter and that the binding is specific, as revealed by siRNA against hypoxia inducible factor-1 and chromatin immunoprecipitation assays. Thus, the activation of DDX3 expression during hypoxia is due to the direct binding of hypoxia inducible factor-1 to hypoxia responsive elements in the DDX3 promoter. In addition, we observed a significant overlap in the protein expression pattern of hypoxia inducible factor-1α and DDX3 in MDA-MB-231 xenograft tumors. Taken together, our results demonstrate, for the first time, the role of DDX3 as a hypoxia-inducible gene that exhibits enhanced expression through the interaction of hypoxia inducible factor-1 with hypoxia inducible factor-1 responsive elements in its promoter region.
- Published
- 2011
- Full Text
- View/download PDF
37. Novel, Broad Spectrum Anti-Cancer Agents Containing the Tricyclic 5:7:5-Fused Diimidazodiazepine Ring System.
- Author
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Kondaskar A, Kondaskar S, Kumar R, Fishbein JC, Muvarak N, Lapidus RG, Sadowska M, Edelman MJ, Bol GM, Vesuna F, Raman V, and Hosmane RS
- Abstract
Synthesis of a series of novel, broad-spectrum anti-cancer agents containing the tricyclic 5:7:5-fused diimidazo[4,5-d:4',5'-f][1,3]diazepine ring system is reported. Compounds 1, 2, 8, 11, and 12 in the series show promising in vitro antitumor activity with low micromolar IC(50)'s against prostate, lung, breast, and ovarian cancer cell lines. Some notions about structure-activity relationships and a possible mechanism of biological activity are presented. Also presented are preliminary in vivo toxicity studies of 1 using SCID mice.
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- 2010
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38. Twist modulates breast cancer stem cells by transcriptional regulation of CD24 expression.
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Vesuna F, Lisok A, Kimble B, and Raman V
- Subjects
- Aldehyde Dehydrogenase genetics, Aldehyde Dehydrogenase metabolism, Aldehyde Dehydrogenase 1 Family, Animals, Binding Sites, Breast Neoplasms genetics, Breast Neoplasms pathology, CD24 Antigen genetics, Cell Line, Cell Line, Tumor, Female, Flow Cytometry, Gene Expression Regulation, Neoplastic, Humans, Hyaluronan Receptors genetics, Hyaluronan Receptors metabolism, Immunoblotting, Isoenzymes genetics, Isoenzymes metabolism, Mammary Neoplasms, Experimental genetics, Mammary Neoplasms, Experimental metabolism, Mammary Neoplasms, Experimental pathology, Mice, Mice, SCID, Neoplastic Stem Cells pathology, Nuclear Proteins genetics, Promoter Regions, Genetic genetics, Protein Binding, RNA Interference, Retinal Dehydrogenase, Transcription, Genetic, Transplantation, Heterologous, Twist-Related Protein 1 genetics, Breast Neoplasms metabolism, CD24 Antigen metabolism, Neoplastic Stem Cells metabolism, Nuclear Proteins metabolism, Twist-Related Protein 1 metabolism
- Abstract
The cancer stem cell paradigm postulates that dysregulated tissue-specific stem cells or progenitor cells are precursors for cancer biogenesis. Consequently, identifying cancer stem cells is crucial to our understanding of cancer progression and for the development of novel therapeutic agents. In this study, we demonstrate that the overexpression of Twist in breast cells can promote the generation of a breast cancer stem cell phenotype characterized by the high expression of CD44, little or no expression of CD24, and increased aldehyde dehydrogenase 1 activity, independent of the epithelial-mesenchymal transition. In addition, Twist-overexpressing cells exhibit high efflux of Hoechst 33342 and Rhodamine 123 as a result of increased expression of ABCC1 (MRP1) transporters, a property of cancer stem cells. Moreover, we show that transient expression of Twist can induce the stem cell phenotype in multiple breast cell lines and that decreasing Twist expression by short hairpin RNA in Twist-overexpressing transgenic cell lines MCF-10A/Twist and MCF-7/Twist as well as in MDA-MB-231 partially reverses the stem cell molecular signature. Importantly, we show that inoculums of only 20 cells of the Twist-overexpressing CD44(+)/CD24(-/low) subpopulation are capable of forming tumors in the mammary fat pad of severe combined immunodeficient mice. Finally, with respect to mechanism, we provide data to indicate that Twist transcriptionally regulates CD24 expression in breast cancer cells. Taken together, our data demonstrate the direct involvement of Twist in generating a breast cancer stem cell phenotype through down-regulation of CD24 expression and independent of an epithelial-mesenchymal transition.
- Published
- 2009
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39. Peptides derived from type IV collagen, CXC chemokines, and thrombospondin-1 domain-containing proteins inhibit neovascularization and suppress tumor growth in MDA-MB-231 breast cancer xenografts.
- Author
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Koskimaki JE, Karagiannis ED, Rosca EV, Vesuna F, Winnard PT Jr, Raman V, Bhujwalla ZM, and Popel AS
- Subjects
- 3T3 Cells, Amino Acid Sequence, Animals, Breast Neoplasms blood supply, Breast Neoplasms pathology, Cell Line, Tumor, Cell Proliferation drug effects, Chemokines, CXC chemistry, Collagen Type IV chemistry, Female, Humans, Immunohistochemistry, Mammary Neoplasms, Experimental blood supply, Mammary Neoplasms, Experimental pathology, Mammary Neoplasms, Experimental prevention & control, Mice, Mice, SCID, Molecular Sequence Data, Neovascularization, Pathologic metabolism, Oligopeptides chemical synthesis, Platelet Endothelial Cell Adhesion Molecule-1 metabolism, Thrombospondins chemistry, Tumor Burden drug effects, Breast Neoplasms prevention & control, Neovascularization, Pathologic prevention & control, Oligopeptides pharmacology, Xenograft Model Antitumor Assays
- Abstract
Angiogenesis or neovascularization, the process of new blood vessel formation from preexisting microvasculature, involves interactions among several cell types including parenchymal, endothelial cells, and immune cells. The formation of new vessels is tightly regulated by a balance between endogenous proangiogenic and antiangiogenic factors to maintain homeostasis in tissue; tumor progression and metastasis in breast cancer have been shown to be angiogenesis-dependent. We previously introduced a systematic methodology to identify putative endogenous antiangiogenic peptides and validated these predictions in vitro in human umbilical vein endothelial cell proliferation and migration assays. These peptides are derived from several protein families including type IV collagen, CXC chemokines, and thrombospondin-1 domain-containing proteins. On the basis of the results from the in vitro screening, we have evaluated the ability of one peptide selected from each family named pentastatin-1, chemokinostatin-1, and properdistatin, respectively, to suppress angiogenesis in an MDA-MB-231 human breast cancer orthotopic xenograft model in severe combined immunodeficient mice. Peptides were administered intraperitoneally once per day. We have demonstrated significant suppression of tumor growth in vivo and subsequent reductions in microvascular density, indicating the potential of these peptides as therapeutic agents for breast cancer.
- Published
- 2009
- Full Text
- View/download PDF
40. Twist is a transcriptional repressor of E-cadherin gene expression in breast cancer.
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Vesuna F, van Diest P, Chen JH, and Raman V
- Subjects
- Cell Line, Tumor, Humans, Breast Neoplasms metabolism, Cadherins metabolism, Gene Expression Regulation, Neoplastic, Nuclear Proteins metabolism, Repressor Proteins metabolism, Transcriptional Activation, Twist-Related Protein 1 metabolism
- Abstract
Twist is a basic helix loop helix protein that plays a role both in human development and in cancer biogenesis. While characterizing the effects of Twist on breast epithelial cell transformation, we identified E-cadherin as a target gene that is down-regulated by Twist. In this study, we demonstrate that Twist can transcriptionally repress E-cadherin in breast cancer cells. Using transient promoter assays, we show that Twist can down-regulate E-cadherin promoter activity by up to two folds. This is further supported by immunoblot analyses which indicates that over-expression of Twist decreases E-cadherin protein levels in breast cancer cell lines. Subsequently, chromatin immunoprecipitation performed on MCF-7/Twist and Hs578 T (high level of endogenous Twist expression) confirmed Twist binding to the E-cadherin promoter. Finally, the functional relevance of this regulation was verified by quantitative real-time PCR and immunohistochemistry on a cohort of breast cancer samples.
- Published
- 2008
- Full Text
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41. Hypoxia regulates choline kinase expression through hypoxia-inducible factor-1 alpha signaling in a human prostate cancer model.
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Glunde K, Shah T, Winnard PT Jr, Raman V, Takagi T, Vesuna F, Artemov D, and Bhujwalla ZM
- Subjects
- Animals, Cell Hypoxia, Choline metabolism, Chromatin Immunoprecipitation, Green Fluorescent Proteins analysis, Green Fluorescent Proteins genetics, Humans, Male, Mice, Mice, SCID, Prostatic Neoplasms genetics, Response Elements, Signal Transduction, Choline Kinase genetics, Gene Expression Regulation, Enzymologic, Gene Expression Regulation, Neoplastic, Hypoxia-Inducible Factor 1, alpha Subunit metabolism, Prostatic Neoplasms enzymology
- Abstract
The intensity of the total choline (tCho) signal in spectroscopic images of tumors is spatially heterogeneous. The likewise heterogeneous physiologic tumor microenvironment may contribute to this heterogeneity. We therefore investigated the relationship between hypoxia, choline metabolites, and choline kinase (Chk) in a human prostate cancer model. Human PC-3 prostate cancer cells were engineered to express enhanced green fluorescent protein (EGFP) under hypoxic conditions. These PC-3-5HRE-EGFP cells were characterized in culture and as tumors transplanted in mice using (1)H magnetic resonance spectroscopy (MRS) and MRS imaging (MRSI) combined with EGFP fluorescence microscopy and imaging. Hypoxic EGFP-fluorescing tumor regions colocalized with regions of high tCho in combined MRSI and optical imaging studies. Cellular phosphocholine (PC) and tCho concentrations as well as Chk expression levels significantly increased following exposure of PC-3 cells to hypoxia. A putative promoter region located 5' of the translation start site of the human chk-alpha gene was cloned and luciferase (Luc)-based reporter vector constructs were generated. Luc reporter assays provided evidence that some of the putative hypoxia response elements (HRE) within this putative chk-alpha promoter region functioned in vitro. Chromatin immunoprecipitation assays using an antibody against hypoxia-inducible factor (HIF)-1 alpha showed that HIF-1 can directly bind this region of the endogenous chk-alpha promoter in hypoxic PC-3-5HRE-EGFP cells. These data suggest that HIF-1 activation of HREs within the putative chk-alpha promoter region can increase Chk-alpha expression within hypoxic environments, consequently increasing cellular PC and tCho levels within these environments.
- Published
- 2008
- Full Text
- View/download PDF
42. Contributing factors of temozolomide resistance in MCF-7 tumor xenograft models.
- Author
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Kato Y, Okollie B, Raman V, Vesuna F, Zhao M, Baker SD, Bhujwalla ZM, and Artemov D
- Subjects
- Animals, Cell Line, Tumor, Dacarbazine pharmacology, Drug Delivery Systems, Female, Humans, Magnetic Resonance Imaging, Mice, Mice, SCID, Models, Biological, Neoplasm Transplantation, O(6)-Methylguanine-DNA Methyltransferase metabolism, Temozolomide, Time Factors, Vascular Endothelial Growth Factor A metabolism, Dacarbazine analogs & derivatives, Drug Resistance, Neoplasm
- Abstract
Vasculature mediated drug resistance in tumors was studied in female SCID mice bearing wild type MCF-7 and adriamycin resistant MCF-7/ADR xenograft using temozolomide (TMZ). A strong tumor growth inhibitory effect of TMZ treatment was observed in MCF-7 tumors during the initial treatment phase with subsequent relapse, but not in MCF-7/ADR tumors. Non-invasive MRI measurements of tumor vascular volume and vascular permeability-surface area product (PS) demonstrated significant reduction of PS in long-term treated MCF-7, but not in MCF-7/ADR tumors. O(6)-Methylguanine-DNA methyltransferase (MGMT) mRNA, and VEGF expression was analyzed using real-time RT-PCR and ELISA, respectively. No significant changes in MGMT mRNA and VEGF expression were observed in either MCF-7 or MCF-7/ADR tumors. However, in vitro incubation of MCF-7 cells with TMZ did induce the expression of MGMT mRNA. In addition, p53 and p21 levels were scored by immunoblotting. Exposure of cells to TMZ did not affect either the p21 or the p53 expression in both MCF-7 and MCF-7/ADR cells. The absence of these molecular responses to TMZ treatment in MCF-7 tumors in vivo supports the possibility that the onset of cancer drug resistance is associated with reduced PS, which can decrease delivery of the drug to cancer cells.
- Published
- 2007
- Full Text
- View/download PDF
43. Histamine: a potential therapeutic agent for breast cancer treatment?
- Author
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Vesuna F and Raman V
- Subjects
- Breast Neoplasms pathology, Breast Neoplasms physiopathology, Cell Line, Tumor, Female, Humans, Signal Transduction drug effects, Signal Transduction physiology, Breast Neoplasms drug therapy, Histamine physiology, Histamine therapeutic use
- Published
- 2006
- Full Text
- View/download PDF
44. Twist overexpression promotes chromosomal instability in the breast cancer cell line MCF-7.
- Author
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Vesuna F, Winnard P Jr, Glackin C, and Raman V
- Subjects
- Aneuploidy, Breast Neoplasms metabolism, Breast Neoplasms ultrastructure, Cell Line, Tumor, Female, Humans, Nucleic Acid Hybridization, Spectral Karyotyping, Breast Neoplasms genetics, Chromosomal Instability, Nuclear Proteins metabolism, Twist-Related Protein 1 metabolism
- Published
- 2006
- Full Text
- View/download PDF
45. HOXA5 regulates hMLH1 expression in breast cancer cells.
- Author
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Duriseti S, Winnard PT Jr, Mironchik Y, Vesuna F, Raman A, and Raman V
- Subjects
- Adaptor Proteins, Signal Transducing, Cell Line, Tumor, Homeodomain Proteins metabolism, Humans, MutL Protein Homolog 1, Oligonucleotide Array Sequence Analysis, Phenotype, Promoter Regions, Genetic, Protein Binding, Saccharomyces cerevisiae, Transcription Factors metabolism, Transcriptional Activation, Breast Neoplasms metabolism, Carrier Proteins biosynthesis, Gene Expression Regulation, Neoplastic, Homeodomain Proteins physiology, Nuclear Proteins biosynthesis
- Abstract
Homeobox protein HOXA5 functions as a transcriptional factor for genes that are not only involved in segmentation identity but also in cell differentiation. Although HOXA5 has been shown to regulate the expression of the tumor-suppressor protein p53, its role in breast tumorigenesis is not well understood. Using yeast as a model system, we now demonstrate that overexpression of HOXA5 in yeast can be used to identify downstream target genes that are homologous in humans. One such identified gene was that of the mismatch repair pathway component MutL homolog 1. Analysis of the promoter region of the gene for human MutL homolog 1 (hMLH1) displayed several putative HOXA5-binding sites. In transient transfection experiments, the overexpression of HOXA5 transactivated the hMLH1 promoter-reporter construct. In addition, chromatin immunoprecipitation assay using a human breast cancer cell line MCF-7 demonstrated that HOXA5 binds to the hMLH1 promoter in vivo. Furthermore, we demonstrate that, in the presence of HOXA5, there is an increase in in vivo repair activity in MCF-7 cells. Taken together, our results indicate that HOXA5 is a transcriptional regulator of hMLH1 in breast cancer cells.
- Published
- 2006
- Full Text
- View/download PDF
46. Twist overexpression induces in vivo angiogenesis and correlates with chromosomal instability in breast cancer.
- Author
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Mironchik Y, Winnard PT Jr, Vesuna F, Kato Y, Wildes F, Pathak AP, Kominsky S, Artemov D, Bhujwalla Z, Van Diest P, Burger H, Glackin C, and Raman V
- Subjects
- Animals, Breast Neoplasms metabolism, Breast Neoplasms pathology, Capillary Permeability physiology, Cell Growth Processes physiology, Cell Line, Tumor, Cell Movement physiology, Claudins, Female, Humans, Membrane Proteins genetics, Mesoderm metabolism, Mice, Mice, SCID, Neoplasm Invasiveness, Neoplasm Transplantation, Neovascularization, Pathologic genetics, Neovascularization, Pathologic metabolism, Neovascularization, Pathologic pathology, Transcriptional Activation, Transplantation, Heterologous, Breast Neoplasms blood supply, Breast Neoplasms genetics, Chromosomal Instability, Nuclear Proteins biosynthesis, Twist-Related Protein 1 biosynthesis
- Abstract
Aggressive cancer phenotypes are a manifestation of many different genetic alterations that promote rapid proliferation and metastasis. In this study, we show that stable overexpression of Twist in a breast cancer cell line, MCF-7, altered its morphology to a fibroblastic-like phenotype, which exhibited protein markers representative of a mesenchymal transformation. In addition, it was observed that MCF-7/Twist cells had increased vascular endothelial growth factor (VEGF) synthesis when compared with empty vector control cells. The functional changes induced by VEGF in vivo were analyzed by functional magnetic resonance imaging (MRI) of MCF-7/Twist-xenografted tumors. MRI showed that MCF-7/Twist tumors exhibited higher vascular volume and vascular permeability in vivo than the MCF-7/vector control xenografts. Moreover, elevated expression of Twist in breast tumor samples obtained from patients correlated strongly with high-grade invasive carcinomas and with chromosome instability, particularly gains of chromosomes 1 and 7. Taken together, these results show that Twist overexpression in breast cancer cells can induce angiogenesis, correlates with chromosomal instability, and promotes an epithelial-mesenchymal-like transition that is pivotal for the transformation into an aggressive breast cancer phenotype.
- Published
- 2005
- Full Text
- View/download PDF
47. Enhanced green fluorescent protein as an alternative control reporter to Renilla luciferase.
- Author
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Vesuna F, Winnard P Jr, and Raman V
- Subjects
- Genes, Reporter genetics, Green Fluorescent Proteins genetics, Luciferases, Renilla genetics, Transfection methods
- Published
- 2005
- Full Text
- View/download PDF
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