Marian L. Waterman
Professor, Microbiology & Molecular Genetics
School of Medicine
School of Medicine
Director, Cancer Research Institute
Deputy Director, Chao Family Comprehensive Cancer Center
Co-Director of Center for Cancer Systems Biology
PH.D., University of California, San Diego
ORCiD: 0000-0003-4823-4968
University of California, Irvine
Rm B251 Medical Sciences I
School of Medicine
Mail Code: 4025
Irvine, CA 92697
Rm B251 Medical Sciences I
School of Medicine
Mail Code: 4025
Irvine, CA 92697
Research Interests
Wnt signaling, intestinal cancer, intestinal stem cells, cancer, gene expression, development, metabolism; intestine; stem cells; pluripotency; LEF and TCF transcription factors; Tumor Heterogeneity; Systems Biology
Websites
Academic Distinctions
American Cancer Society Postdoctoral Fellowship
Special Fellow, Leukemia Society
Special Fellow, Leukemia Society
Appointments
Post-Doctoral Fellow, The Salk Institute, 1987-1993
Research Abstract
OVERVIEW
The goal of research in the Waterman laboratory is to understand how the Wnt signaling pathway regulates target genes through the actions of LEF/TCF transcription factors. Dysfunctional Wnt signals misregulate gene expression and cause cancer and most of our work on LEF/TCFs is within the context of the final step of this dysregulated pathway. LEF/TCF proteins bind to Wnt Response Elements in target genes and recruit ß-catenin for gene activation or repressors for gene silencing. There are two main areas of research: 1) Regulation of LEF/TCF expression, and 2) Structure and activities of LEF/TCFs. Our work in these two areas has led to new insights on LEF/TCF structure, Wnt response element recognition, signals from the tumor microenvironment and chromatin regulators that modify the types and amounts of LEF/TCFs. In almost every aspect LEF/TCF expression and/or function has been distorted in cancer.
REGULATION OF LEF/TCF TRANSCRIPTION AND TRANSLATION
Transcription: The LEF1 locus has two promoters for RNA polymerase II transcription, where each promoter produces a differently acting form of LEF-1. One promoter is activated in Wnt-linked cancers (P1), the other promoter is silenced (P2). We aim to identify the regulatory elements, transcription factors and signaling pathways that create this pattern, as the activated-silenced pattern produces only Wnt-promoting forms of LEF-1. We are also working to define common themes of transcription regulation of all four mammalian LEF/TCFs for a more comprehensive view of expression of this family is modulated during development and disease.
Translation: LEF1 mRNA produced from P1 contains two Internal Ribosome Entry Sites in its long 5’ UTR. We have discovered a cancer-relevant signaling pathway that regulates IRES activity and LEF-1 protein production. Current efforts are focused on a new method to isolate LEF1 mRNA/protein complexes for mass spectrometry analysis and IRES-factor discovery.
STRUCTURE AND ACTIVITIES OF LEF/TCFS
A new DNA binding domain: We have discovered a second DNA binding domain in the C-terminus of TCFs. Previously known as the CRARF region, we refer to it as the “C-clamp” because of four invariant and essential cysteines. The C-clamp is a new type of DNA binding domain and appears to be critical for TCF-mediated growth control. We aim to define the role of this domain in carcinogenesis and obtain structural and gene targeting information to develop ways to interfere with its function.
Nuclear export of TCFs: Another major effort in the laboratory is a focus on subcellular trafficking of LEF/TCFs. Our early research identified nuclear localization activities and receptors that ushered LEF/TCFs into the nucleus. More recently, we have discovered that certain mammalian TCFs engage in nuclear/cytoplasmic shuttling – an activity that is active and distorted in cancer. We have discovered a Wnt-linked kinase cascade that can control this shuttling, linking TCF trafficking to extracellular signals that may be aberrant in the tumor microenvironment. The Wnt-linked signal has elements of “canonical” and “non-canonical” signaling, and we are working to define these novel elements as well as the molecular changes in LEF/TCFs when export is triggered.
The goal of research in the Waterman laboratory is to understand how the Wnt signaling pathway regulates target genes through the actions of LEF/TCF transcription factors. Dysfunctional Wnt signals misregulate gene expression and cause cancer and most of our work on LEF/TCFs is within the context of the final step of this dysregulated pathway. LEF/TCF proteins bind to Wnt Response Elements in target genes and recruit ß-catenin for gene activation or repressors for gene silencing. There are two main areas of research: 1) Regulation of LEF/TCF expression, and 2) Structure and activities of LEF/TCFs. Our work in these two areas has led to new insights on LEF/TCF structure, Wnt response element recognition, signals from the tumor microenvironment and chromatin regulators that modify the types and amounts of LEF/TCFs. In almost every aspect LEF/TCF expression and/or function has been distorted in cancer.
REGULATION OF LEF/TCF TRANSCRIPTION AND TRANSLATION
Transcription: The LEF1 locus has two promoters for RNA polymerase II transcription, where each promoter produces a differently acting form of LEF-1. One promoter is activated in Wnt-linked cancers (P1), the other promoter is silenced (P2). We aim to identify the regulatory elements, transcription factors and signaling pathways that create this pattern, as the activated-silenced pattern produces only Wnt-promoting forms of LEF-1. We are also working to define common themes of transcription regulation of all four mammalian LEF/TCFs for a more comprehensive view of expression of this family is modulated during development and disease.
Translation: LEF1 mRNA produced from P1 contains two Internal Ribosome Entry Sites in its long 5’ UTR. We have discovered a cancer-relevant signaling pathway that regulates IRES activity and LEF-1 protein production. Current efforts are focused on a new method to isolate LEF1 mRNA/protein complexes for mass spectrometry analysis and IRES-factor discovery.
STRUCTURE AND ACTIVITIES OF LEF/TCFS
A new DNA binding domain: We have discovered a second DNA binding domain in the C-terminus of TCFs. Previously known as the CRARF region, we refer to it as the “C-clamp” because of four invariant and essential cysteines. The C-clamp is a new type of DNA binding domain and appears to be critical for TCF-mediated growth control. We aim to define the role of this domain in carcinogenesis and obtain structural and gene targeting information to develop ways to interfere with its function.
Nuclear export of TCFs: Another major effort in the laboratory is a focus on subcellular trafficking of LEF/TCFs. Our early research identified nuclear localization activities and receptors that ushered LEF/TCFs into the nucleus. More recently, we have discovered that certain mammalian TCFs engage in nuclear/cytoplasmic shuttling – an activity that is active and distorted in cancer. We have discovered a Wnt-linked kinase cascade that can control this shuttling, linking TCF trafficking to extracellular signals that may be aberrant in the tumor microenvironment. The Wnt-linked signal has elements of “canonical” and “non-canonical” signaling, and we are working to define these novel elements as well as the molecular changes in LEF/TCFs when export is triggered.
Awards and Honors
2018 Alumni Fellow Honoree, Oregon State University
Publications
Lyou, Y., A.N. Habowski, G.T. Chen and M.L. Waterman. 2017. Inhibition of Nuclear Wnt Signaling: Challenges of an Elusive Target for Cancer Therapy. Br J Pharmacol.
174(24):4589-4599. doi: 10.1111/bph.13963
174(24):4589-4599. doi: 10.1111/bph.13963
Chen, G.T. and M.L. Waterman. 2015. Cancer: Leaping the E-cadherin Hurdle. 2015. EMBO J. 34:2307-9. PMC457051
Habowski, A. N., J. M. Bates, J.L. Flesher, R. A. Edwards, M. L. Waterman. 2020. Isolation of murine large intestinal crypt cell populations with flow sorting. Nature Protocol Exchange. DOI:10.21203/rs.3.pex-994/v1
Habowski, A.N., J.L. Flesher, J.M. Bates, C.F. Tsai, K. Martin, R. Zhao, A.K. Ganesan, R.A. Edwards, T. Shi, H.S.Wiley, Y. Shi, K.J. Hertel, and M.L. Waterman. 2020. Transcriptomic and proteomic signatures of stemness and differentiation in the colon crypt. Commun Biol. 19:3(1):453. doi: 10.1038/s42003-020-01181-z. PMID: 32814826
Tran, T.Q., E.A. Hanse, A.N. Habowski, H. Li, M.B. Ishak Gabra, Y. Yang, X.H. Lowman, A.M. Ooi, S.Y. Liao, R.A. Edwards, M.L. Waterman & M. Kong. 2020. a-Ketoglutarate attenuates Wnt signaling and drives differentiation in colorectal cancer. Nature Cancer 1: 345–358. doi:10.1038/s43018-020-0035-5
Morgan, R. B., J. Ridsdale, M. Payne, K. Heesom, M.C. Wilson, A. Davidson, A. Greenhough, S. Davies, A. C. Williams, A. Blair, M.L. Waterman, A. Tonks, R. L. Darley. 2019. LEF-1 Drives Aberrant ?-Catenin Nuclear Localization In Myeloid Leukemia Cells. Haematologica 104:1365-1377. doi:10.3324/haematol.2018.202846. PMC6601079
Sierra, R.A., N. P. Hoverter, R. N. Ramirez, Linh M. Vuong, A. Mortazavi, B. J. Merrill, M.L. Waterman*, and P. J. Donovan*. 2018. TCF7L1 suppresses primitive streak gene expression to support human embryonic stem cell pluripotency. Development, 145(4):dev161075. doi: 10.1242/dev.161075. PMC5869011
Lee, M. ‡, G. T. Chen‡, E. Puttock, K. Wang, R. A. Edwards, M. L. Waterman*, and J. Lowengrub*. 2017. Mathematical modeling links Wnt signaling to emergent patterns of metabolism in colon cancer. Molecular Systems Biology 13:912. PMC5327728
Sprowl-Tanio, S., A. Habowski, Y. Lyou, K. T. Pate, M. M. McQuade, F. Grun, and M.L. Waterman. 2016. Lactate/Pyruvate Transporter MCT-1 is a direct Wnt target that confers sensitivity to 3 bromopyruvate in colon cancer. Cancer & Metabolism 4:20. PMC5046889
Vuong L.M., K. Chellappa, J.M. Dhahbi, J.R. Deans, B. Fang, E. Bolotin, N.V. Titova, N.P. Hoverter, S.R. Spindler, M.L. Waterman, and F.M. Sladek. 2015 Differential Effects of Hepatocyte Nuclear Factor 4a Isoforms on Tumor Growth and T-Cell Factor 4/AP-1 Interactions in Human Colorectal Cancer Cells. 2015. Mol Cell Biol. 35:3471-90. PMC4573706
Kurimoto S, Jung J, Tapadia M, Lengfeld J, Agalliu D, Waterman M, Mozaffar T, Gupta R. 2015. Activation of the Wnt/ß-catenin signaling cascade after traumatic nerve injury. Neuroscience. 3:294:101-108. PMC5384639
Hoverter, N.P., M. D. Zeller, M. M. McQuade, A. Garibaldi, A. Busch, E. M. Selwan, K. J. Hertel, P. Baldi, and M.L. Waterman. 2014. The TCF C-clamp DNA Binding Domain Expands the Wnt Transcriptome via Alternative Target Recognition. Nucleic Acids Res. 42(22):13615-32
MacDonald B.T., A. Hien, X. Zhang, O. Iranloye, D. M. Virshup, M. L. Waterman, and X. He. 2014. Disulfide Bond Requirements for Active Wnt Ligands. J. Biol. Chem. 289:18122-18136. PMC4140276
Konstorum, A., S.A. Sprowl, M.L.Waterman, A.D. Lander, J.S. Lowengrub. 2013. Predicting mechanism of biphasic growth factor action on tumor growth using a multi-species model with feedback control. J Coupled Syst Multiscale Dyn. 1(4):459-467. PMC4112130
Cheng, Y., A.K. Cheung, J.M. Ko., Y.P. Phoon, P.M. Chiu, P.H. Lo, M.L. Waterman, M.L. Lung. 2013. Physiological beta-catenin signaling controls self-renewal networks and generation of stem-like cells from nasopharyngeal carcinoma. BMC Cell Biol., 14:44. PMC3819748
Stringari, C., R.A. Edwards, K.T. Pate, M.L. Waterman, P.J. Donovan, E. Gratton. 2012. Metabolic trajectory of cellular differentiation in small intestine by Phasor Fluorescence Lifetime Microscopy of NADH. Sci. Rep. 2:568. PMC3416911
Najdi, R., K. Proffitt, S. Sprowl, S. Kaur, J. Yu, T.M. Covey, D.M. Virshup*, and M.L. Waterman*. 2012. A uniform human Wnt expression library reveals a shared secretory pathway and unique signaling activities. Differentiation, 84(2):203-213. PMC4015730
Hoverter, N.P., J.-H. Ting, S. Sundaresh, P. Baldi, M.L. Waterman. 2012. A WNT/p21 circuit directed by the C-clamp, a sequence-specific DNA binding domain in TCFs. Mol. Cell Biol., 32:3648-3662. PMC3430198
Tsai, B.P., X. Wang, L. Huang, M.L. Waterman. 2011. Quantitative profiling of in vivo-assembled RNA-protein complexes using a novel integrated proteomic approach. Mol Cell Proteomics, 10(4):M110.007385.
Tsai, B.P., N.P. Hoverter, and M.L. Waterman. 2012. Blending hippo and WNT: sharing messengers and regulation. Cell, 151:1401-3.
Cadigan, K.M., and M.L. Waterman. 2012. TCF/LEFs and Wnt signaling in the nucleus. Perspectives in Biology (Cold Spring Harbor). 4: doi:pii: a007906
Najdi, R., R. Holcombe, and M.L. Waterman. 2011. Wnt signaling and colon carcinogenesis: Beyond APC. J. Carcinogenesis, 10:5-10.
Semler, B. L., and M. L. Waterman. 2008. IRES-mediated pathways to polysomes: nuclear versus cytoplasmic routes. Trends Microbiol.,16:1-5.
Arce, L., N.N. Yokoyama and M.L. Waterman. 2006. Diversity of LEF/TCF action in development and disease. Oncogene 25:7492-7502.
Semler, B. L., and M. L. Waterman. 2008. IRES-mediated pathways to polysomes: nuclear versus cytoplasmic routes. Trends Microbiol.,16:1-5.
Lyou, Y., A.N. Habowski, G.T. Chen and M.L. Waterman. 2017. Inhibition of Nuclear Wnt Signaling: Challenges of an Elusive Target for Cancer Therapy. Br J Pharmacol. 174(24):4589-4599. doi:10.1111/bph.13963.
Habowski, A.N., J.L. Flesher, J.M. Bates, C.F. Tsai, K. Martin, R. Zhao, A.K. Ganesan, R.A. Edwards, T. Shi, H.S.Wiley, Y. Shi, K.J. Hertel, and M.L. Waterman. 2020. Transcriptomic and proteomic signatures of stemness and differentiation in the colon crypt. Commun Biol. 19:3(1):453. doi: 10.1038/s42003-020-01181-z. PMID: 32814826
Kurimoto S, Jung J, Tapadia M, Lengfeld J, Agalliu D, Waterman M, Mozaffar T, Gupta R. 2015. Activation of the Wnt/ß-catenin signaling cascade after traumatic nerve injury. Neuroscience. 3:294:101-108. PMC5384639
Vuong L.M., K. Chellappa, J.M. Dhahbi, J.R. Deans, B. Fang, E. Bolotin, N.V. Titova, N.P. Hoverter, S.R. Spindler, M.L. Waterman, and F.M. Sladek. 2015 Differential Effects of Hepatocyte Nuclear Factor 4a Isoforms on Tumor Growth and T-Cell Factor 4/AP-1 Interactions in Human Colorectal Cancer Cells. 2015. Mol Cell Biol. 35:3471-90.
Sprowl-Tanio, S., A. Habowski, Y. Lyou, K. T. Pate, M. M. McQuade, F. Grun, and M.L. Waterman. 2016. Lactate/Pyruvate Transporter MCT-1 is a direct Wnt target that confers sensitivity to 3 bromopyruvate in colon cancer. Cancer & Metabolism 4:20. PMC5046889
Lee, M. ‡, G. T. Chen‡, E. Puttock, K. Wang, R. A. Edwards, M. L. Waterman*, and J. Lowengrub*. 2017. Mathematical modeling links Wnt signaling to emergent patterns of metabolism in colon cancer. Molecular Systems Biology 13:912. PMC5327728. doi: 10.15252/msb.20167386.
Sierra, R.A., N. P. Hoverter, R. N. Ramirez, Linh M. Vuong, A. Mortazavi, B. J. Merrill, M.L. Waterman*, and P. J. Donovan*. 2018. TCF7L1 suppresses primitive streak gene expression to support human embryonic stem cell pluripotency. Development, 145(4):dev161075. doi: 10.1242/dev.161075. PMC5869011
Morgan, R. B., J. Ridsdale, M. Payne, K. Heesom, M.C. Wilson, A. Davidson, A. Greenhough, S. Davies, A. C. Williams, A. Blair, M.L. Waterman, A. Tonks, R. L. Darley. 2019. LEF-1 Drives Aberrant Beta-Catenin Nuclear Localization In Myeloid Leukemia Cells. Haematologica 104:1365-1377. doi:10.3324/haematol.2018.202846. PMC6601079
Tran, T.Q., E.A. Hanse, A.N. Habowski, H. Li, M.B. Ishak Gabra, Y. Yang, X.H. Lowman, A.M. Ooi, S.Y. Liao, R.A. Edwards, M.L. Waterman & M. Kong. 2020. a-Ketoglutarate attenuates Wnt signaling and drives differentiation in colorectal cancer. Nature Cancer 1: 345–358. doi:10.1038/s43018-020-0035-5.
Hoppler, S. and M.L. Waterman. 2013. Variety of vertebrate TCF/LEF structure, function, and regulation, in: Wnt signaling in Development and Disease: Molecular Mechanisms and Biological Functions (S. Hoppler and R.T. Moon eds.), Wiley Publishers.
Tsai, B.P., J. Jimenez, S. Lim, K.D. Fitzgerald, M. Zhang, C.T.H. Chuah, H. Axelrod, L. Nelson, S. T. Ong, B.L. Semler, and M.L. Waterman. 2014. A Novel Bcr-Abl-mTOR-eIF4A Axis Regulates IRES-Mediated Translation of LEF-1. Open Biology, 4:140180. PMC4248067
Ehsan S.M., K. M. Welch-Reardon, M.L. Waterman, C.C. Hughes, S.C. George. 2014. A three-dimensional in vitro model of tumor cell intravasation. Integr Biol. 6:603-10. PMC4046910
Chodaparambil, J.V., K.T. Pate, M.R.D. Hepla, B.P. Tsai, U.M. Muthurajan, K. Luger, M.L. Waterman, and W.I. Weis. 2014. Molecular Functions of the TLE Tetramerization Domain in Wnt Target Gene Repression. EMBO J, 33:719-31. PMC4000089
Pate, K.T., C. Stringari, S. Sprowl-Tanio, K. Wang, T. TeSlaa, N.P. Hoverter, M.M. McQuade, C. Garner, M.A. Digman, M.A. Teitell, R.A. Edwards, E. Gratton, M.L. Waterman. 2014. WNT Signaling Directs a Metabolic Program of Glycolysis and Angiogenesis in Colon Cancer. EMBO J. 33:1454-73. PMC4194089
Wu*, B., S. Piloto, W. Zeng, N.P. Hoverter, T.F. Schilling, and M.L. Waterman*. 2013. Ring Finger Protein 14 is a novel regulator of TCF/ß-catenin-mediated transcription and colon cancer cell survival. EMBO Rep. 14:347-355. PMC3615654
Atcha, F.A., A. Syed, J.E. Munguia, J-H. T. Ting, J.L. Marsh*, and M.L. Waterman*. 2007. Inclusion of a second DNA binding domain by alternative splicing converts TCF-1 and TCF-4 into stronger Wnt effectors. Mol. Cell. Biol. 27:8352-8363
Sprowl, S. and M.L. Waterman. 2013. Past Visits Present: TCF/LEFs partner with ATFs for ß-catenin independent activity. PLoS Genetics, 9(8):e1003745.
Najdi, R., R. Holcombe, and M.L. Waterman. 2011. Wnt signaling and colon carcinogenesis: Beyond APC. J. Carcinogenesis 10:5.
Yokoyama, N., K.T. Pate, S. Sprowl, and M.L. Waterman. 2010. A role for YY1 repression of dominant negative LEF-1 expression in colon cancer. Nucleic Acids Res. 38:6375-88
Wang, Y., R. Dhopeshwarkar, R. Najdi, M.L. Waterman, C. E. Sims, and N. Allbritton. 2010. Microdevice to capture colon crypts for in vitro studies. Lab on Chip. 10:1596-1603
Sikandar, S. S., K.T. Pate, S. Anderson, D. Dizon, R.A. Edwards, M.L. Waterman and S.M Lipkin. 2009. NOTCH signaling is required for colon cancer initiating cell tumor formation, self-renewal and repression of secretory cell lineage differentiation. Cancer Res. 70:1469-78.
Najdi, R., A. Syed, L. Arce, H. Theisen, J.T. Ting, F. Atcha, R.A. Edwards, J.L. Marsh*, M.L. Waterman*. 2009. Nuclear exclusion of growth-suppressing TCF-1 via a Wnt-mediated signal in colon cancer. Oncogene. 28:4133-46
Arce L., K. T. Pate, M.L. Waterman. 2009. Groucho recognizes a peptide motif in LEF-1 for HDAC-dependent repression in a subset of colon cancers. BMC Cancer, 9:159
Hoverter, N.P. and M.L. Waterman. 2008. “A Wnt-fall for gene regulation: repression”. Science Signal. 1:43-46
Yang, M., M.L. Waterman, and R.K. Brachmann. 2008. hADA2 and hADA3 are essential for acetylation, transcriptional activity and proliferative effects of beta-catenin. Cancer Biol. Ther.7: 120-128
Li, B., C. Rheaume, A. Teng, V. Bilanchone, J.E. Munguia, M. Hu, S. Jessen, S. Piccolo, M.L. Waterman, and X. Dai. 2007. Characterization of developmental phenotypes of mice deficient for pygopus 2. Genesis, 45:318-32
Theisen, H., A. Syed, B.T. Nguyen, T. Lukacsovich, J. Purcell, G. P. Srivastava, D. Iron, K. Gaudenz, Q. Nie, R.Y.M. Wan, M.L. Waterman, and J.L. Marsh. 2007. Wingless directly represses DPP morphogen expression via an armadillo/TCF/Brinker complex. PLoS ONE 2:e142
Arce, L., N.N. Yokoyama and M.L. Waterman. 2006. Diversity of LEF/TCF action in development and disease. Oncogene 25:7492-7502
Li, T. W.-H., J.-H. T. Ting, N. Yokoyama, A. Bernstein, M. van de Wetering, and M.L. Waterman. 2006. WNT activation and alternative promoter repression of LEF1 in colon cancer. Mol. Cell. Biol. 26: 5284-5299
Jimenez, J.J., G.M. Jang, B.L. Semler and M.L. Waterman. 2005. An internal ribosome entry site mediates translation of Lymphoid enhancer factor-1. RNA 11:1385-1399
Atcha, F.A., J.E. Munguia., W.-H. Li, K. Hovanes, M.L. Waterman. 2003. A new b-catenin dependent activation domain in T Cell Factor. J Biol Chem 278:16169-16175
Ishitani, T., S. Kishida, J. Hyodo, N. Oeno, J. Yasuda, M. L. Waterman, H. Shibuya, R.T. Moon, J. Ninomiya-Tsuji, and K. Matsumoto. 2003. The TAK1-NLK MAPK cascade functions in the Wnt-5a/Ca+2 pathway to antagonize Wnt/ß-catenin signaling. Mol. Cell Biol. 23:131-139
Hovanes, K.H., J.E. Munguia, T. Truong, T. Milovanovic, T.W.H. Li, J. L. Marsh, R.F. Holcombe and M.L. Waterman. 2001. Selective expression of b-catenin-sensitive forms of Lymphoid Enhancer Factor-1 in Colon Cancer. Nature Genetics 28:53-57.
Hovanes, K.H., T.W.H. Li, and M.L. Waterman. 2000. The human LEF-1 gene contains a promoter preferentially active in lymphocytes and encodes multiple isoforms derived from alternative splicing. Nucleic Acids Res. 28(9):1994-2003.
Professional Societies
AAAS
AACR
Other Experience
Member
American Cancer Society Council for Extramural Grants 2010—2014
American Cancer Society Council for Extramural Grants 2010—2014
Graduate Programs
Cellular and Molecular Biosciences
Research Centers
UCI Chao Family Comprehensive Cancer Center
Center for Complex Biological Systems
Sue and Bill Gross Stem Cell Center
Institute for Immunology
Cancer Research Institute
Center for Cancer Systems Biology
Link to this profile
https://faculty.uci.edu/profile/?facultyId=2406
https://faculty.uci.edu/profile/?facultyId=2406
Last updated
12/10/2020
12/10/2020