TCF12 Deficiency Impairs the Proliferation of Glioblastoma Tumor Cells and Improves Survival
Abstract
:Simple Summary
Abstract
1. Introduction
2. Materials and Methods
2.1. Mice
2.2. Cell Culture
2.3. Western Blot
2.4. Histology and Immunofluorescence and Immunohistochemistry
2.5. Quantitative RT-PCR
2.6. EdU Proliferation Assay
2.7. Generation of CRISPR-Mediated Knockout GL261, U251, and Human CBTP-14 Cell Lines
2.8. MRI of Xenografts
2.9. Statistical Analysis
2.10. RNA Sequencing and Data Analysis
3. Results
3.1. Tcf12 Is Upregulated in Human GBM Cells and Is Highly Expressed in Enriched GSCs
3.2. TCF12 Regulates the Cell Cycle in Glioblastoma Stem Cells
3.3. TCF12 Loss Leads to Upregulation of Signature Genes of the Oligodendrocytic Lineage
3.4. TCF12 Loss Slows Tumor Growth In Vivo and Improves Animal Survival
3.5. Overexpression of TCF12 Leads to More Aggressive Tumors In Vivo
3.6. Identification of Genes and Pathways Regulated by TCF12 in GBM Cells
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
- Hartmann, C.; Hentschel, B.; Wick, W.; Capper, D.; Felsberg, J.; Simon, M.; Westphal, M.; Schackert, G.; Meyermann, R.; Pietsch, T.; et al. Patients with IDH1 wild type anaplastic astrocytomas exhibit worse prognosis than IDH1-mutated glioblastomas, and IDH1 mutation status accounts for the unfavorable prognostic effect of higher age: Implications for classification of gliomas. Acta Neuropathol. 2010, 120, 707–718. [Google Scholar]
- Stupp, R.; Mason, W.; van den Bent, M.; Weller, M.; Fisher, B.; Taphoorn, M.; Belanger, K.; Brandes, A.; Marosi, C.; Bogdahn, U.; et al. Radiotherapy plus concomitant and adjuvant temozolomide for glioblastoma. N. Engl. J. Med. 2005, 352, 987–996. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Omuro, A.; DeAngelis, L.M. Glioblastoma and other malignant gliomas: A clinical review. JAMA 2013, 310, 1842–1850. [Google Scholar] [CrossRef]
- Wang, J.; Cazzato, E.; Ladewig, E.; Frattini, V.; Rosenbloom, D.; Zairis, S.; Abate, F.; Liu, Z.; Elliott, O.; Shin, Y.-J.; et al. Clonal evolution of glioblastoma under therapy. Nat. Genet. 2016, 48, 768–776. [Google Scholar]
- Gimple, R.C.; Bhargava, S.; Dixit, D.; Rich, J.N. Glioblastoma stem cells: Lessons from the tumor hierarchy in a lethal cancer. Genes Dev. 2019, 33, 591–609. [Google Scholar]
- Neftel, C.; Laffy, J.; Filbin, M.; Hara, T.; Shore, M.; Rahme, G.; Richman, A.; Silverbush, D.; Shaw, M.; Hebert, C.; et al. An Integrative Model of Cellular States, Plasticity, and Genetics for Glioblastoma. Cell 2019, 178, 835–849.e21. [Google Scholar]
- Alvarado, A.G.; Turaga, S.M.; Sathyan, P.; Mulkearns-Hubert, E.E.; Otvos, B.; Silver, D.J.; Hale, J.S.; Flavahan, W.A.; Zinn, P.O.; Sinyuk, M.; et al. Coordination of self-renewal in glioblastoma by integration of adhesion and microRNA signaling. Neuro Oncol. 2016, 18, 656–666. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Singh, S.K.; Hawkins, C.; Clarke, I.D.; Squire, J.A.; Bayani, J.; Hide, T.; Henkelman, R.M.; Cusimano, M.D. Identification of human brain tumour initiating cells. Nature 2004, 432, 396–401. [Google Scholar] [CrossRef] [PubMed]
- Lathia, J.D.; Mack, S.C.; Mulkearns-Hubert, E.E.; Valentim, C.L.; Rich, J.N. Cancer stem cells in glioblastoma. Genes Dev. 2015, 29, 1203–1217. [Google Scholar] [CrossRef] [Green Version]
- Heddleston, J.M.; Li, Z.; McLendon, R.E.; Hjelmeland, A.B.; Rich, J.N. The hypoxic microenvironment maintains glioblastoma stem cells and promotes reprogramming towards a cancer stem cell phenotype. Cell Cycle 2009, 8, 3274–3284. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Seidel, S.; Garvalov, B.K.; Wirta, V.; von Stechow, L.; Schänzer, A.; Meletis, K.; Wolter, M.; Sommerlad, D.; Henze, A.-T.; Nistér, M.; et al. A hypoxic niche regulates glioblastoma stem cells through hypoxia inducible factor 2 alpha. Brain 2010, 133, 983–995. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Pine, A.R.; Cirigliano, S.M.; Nicholson, J.G.; Hu, Y.; Linkous, A.; Miyaguchi, K.; Edwards, L.; Singhania, R.; Schwartz, T.H.; Ramakrishna, R.; et al. Tumor Microenvironment Is Critical for the Maintenance of Cellular States Found in Primary Glioblastomas. Cancer Discov. 2020, 10, 964–979. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Suva, M.L.; Tirosh, I. The Glioma Stem Cell Model in the Era of Single-Cell Genomics. Cancer Cell 2020, 37, 630–636. [Google Scholar] [CrossRef] [PubMed]
- Azzarelli, R.; Simons, B.; Philpott, A. The developmental origin of brain tumours: A cellular and molecular framework. Development 2018, 145, dev162693. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Galli, R.; Binda, E.; Orfanelli, U.; Cipelletti, B.; Gritti, A.; De Vitis, S.; Fiocco, R.; Foroni, C.; Dimeco, F.; Vescovi, A. Isolation and characterization of tumorigenic, stem-like neural precursors from human glioblastoma. Cancer Res. 2004, 64, 7011–7021. [Google Scholar] [CrossRef] [Green Version]
- Riddick, G.; Kotliarova, S.; Rodriguez, V.; Kim, H.S.; Linkous, A.; Storaska, A.J.; Ahn, S.; Walling, J.; Belova, G. A Core Regulatory Circuit in Glioblastoma Stem Cells Links MAPK Activation to a Transcriptional Program of Neural Stem Cell Identity. Sci. Rep. 2017, 7, 43605. [Google Scholar] [CrossRef] [Green Version]
- Sherry, M.M.; Reeves, A.; Wu, J.K.; Cochran, B.H. STAT3 is required for proliferation and maintenance of multipotency in glioblastoma stem cells. Stem Cells 2009, 27, 2383–2392. [Google Scholar] [CrossRef] [Green Version]
- Lee, J.H.; Lee, J.E.; Kahng, J.Y.; Kim, S.H.; Park, J.S.; Yoon, S.J.; Um, J.-Y.; Kim, W.K.; Lee, J.-K.; Park, J.; et al. Human glioblastoma arises from subventricular zone cells with low-level driver mutations. Nature 2018, 560, 243–247. [Google Scholar] [CrossRef]
- Prager, B.C.; Bhargava, S.; Mahadev, V.; Hubert, C.G.; Rich, J.N. Glioblastoma Stem Cells: Driving Resilience through Chaos. Trends Cancer 2020, 6, 223–235. [Google Scholar] [CrossRef] [Green Version]
- Kelly, J.J.; Stechishin, O.; Chojnacki, A.; Lun, X.; Sun, B.; Senger, D.L.; Forsyth, P.; Auer, R.N.; Dunn, J.F.; Cairncross, J.G.; et al. Proliferation of human glioblastoma stem cells occurs independently of exogenous mitogens. Stem Cells 2009, 27, 1722–1733. [Google Scholar] [CrossRef]
- Wang, L.H.; Baker, N.E. E Proteins and ID Proteins: Helix-Loop-Helix Partners in Development and Disease. Dev. Cell 2015, 35, 269–280. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Kee, B.L. E and ID proteins branch out. Nat. Rev. Immunol. 2009, 9, 175–184. [Google Scholar] [CrossRef]
- Uittenbogaard, M.; Chiaramello, A. Expression of the bHLH transcription factor Tcf12 (ME1) gene is linked to the expansion of precursor cell populations during neurogenesis. Brain Res. Gene Expr. Patterns 2002, 1, 115–121. [Google Scholar] [CrossRef]
- Shin, J.; Berg, D.; Zhu, Y.; Shin, J.; Song, J.; Bonaguidi, M.; Enikolopov, G.; Nauen, D.; Christian, K.; Ming, G.-l.; et al. Single-Cell RNA-Seq with Waterfall Reveals Molecular Cascades underlying Adult Neurogenesis. Cell Stem Cell 2015, 17, 360–372. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Codega, P.; Silva-Vargas, V.; Paul, A.; Maldonado-Soto, A.R.; DeLeo, A.M.; Pastrana, E.; Doetsch, F. Prospective identification and purification of quiescent adult neural stem cells from their in vivo niche. Neuron 2014, 82, 545–559. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Xie, X.P.; Laks, D.R.; Sun, D.; Poran, A.; Laughney, A.M.; Wang, Z.; Sam, J.; Belenguer, G.; Fariñas, I.; Elemento, O.; et al. High-resolution mouse subventricular zone stem-cell niche transcriptome reveals features of lineage, anatomy, and aging. Proc. Natl. Acad. Sci. USA 2020, 117, 31448–31458. [Google Scholar] [CrossRef] [PubMed]
- Godoy, P.R.; Montaldi, A.; Sakamoto-Hojo, E. HEB silencing induces anti-proliferative effects on U87MG cells cultured as neurospheres and monolayers. Mol. Med. Rep. 2016, 14, 5253–5260. [Google Scholar] [CrossRef] [Green Version]
- Hambardzumyan, D.; Parada, L.F.; Holland, E.C.; Charest, A.L. Genetic modeling of gliomas in mice: New tools to tackle old problems. Glia 2011, 59, 1155–1168. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Huse, J.T.; Holland, E.C. Genetically engineered mouse models of brain cancer and the promise of preclinical testing. Brain Pathol. 2009, 19, 132–143. [Google Scholar] [CrossRef] [Green Version]
- Wee, B.; Charles, N.; Holland, E. Animal models to study cancer-initiating cells from glioblastoma. Front. Biosci. (andmark Ed.) 2011, 16, 2243–2258. [Google Scholar] [CrossRef] [Green Version]
- Bertoli, C.; Skotheim, J.; de Bruin, R. Control of cell cycle transcription during G1 and S phases. Nat. Rev. Mol. Cell. Biol. 2013, 14, 518–528. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Tian, R.; Wang, J.; Yan, H.; Wu, J.; Xu, Q.; Zhan, X.; Gui, Z.; Ding, M.; He, J. Differential expression of miR16 in glioblastoma and glioblastoma stem cells: Their correlation with proliferation, differentiation, metastasis and prognosis. Oncogene 2017, 36, 5861–5873. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Juan, G.; Traganos, F.; James, W.M.; Ray, J.M.; Roberge, M.; Sauve, D.M.; Anderson, H.; Darzynkiewicz, Z. Histone H3 phosphorylation and expression of cyclins A and B1 measured in individual cells during their progression through G2 and mitosis. Cytometry 1998, 32, 71–77. [Google Scholar] [CrossRef]
- Wesseling, P.; van den Bent, M.; Perry, A. Oligodendroglioma: Pathology, molecular mechanisms and markers. Acta Neuropathol. 2015, 129, 809–827. [Google Scholar] [CrossRef] [Green Version]
- Labreche, K.; Simeonova, I.; Kamoun, A.; Gleize, V.; Chubb, D.; Letouzé, E.; Riazalhosseini, Y.; Dobbins, S.E.; Elarouci, N.; Ducray, F.; et al. TCF12 is mutated in anaplastic oligodendroglioma. Nat. Commun. 2015, 6, 7207. [Google Scholar] [CrossRef] [Green Version]
- Louis, D.N.; Perry, A.; Wesseling, P.; Brat, D.; Cree, I.; Figarella-Branger, D.; Hawkins, C.; Ng, H.K.; Pfister, S.; Reifenberger, G.; et al. The 2021 WHO Classification of Tumors of the Central Nervous System: A summary. Neuro. Oncol. 2021, 23, 1231–1251. [Google Scholar] [CrossRef]
Disclaimer/Publisher’s Note: The statements, opinions and data contained in all publications are solely those of the individual author(s) and contributor(s) and not of MDPI and/or the editor(s). MDPI and/or the editor(s) disclaim responsibility for any injury to people or property resulting from any ideas, methods, instructions or products referred to in the content. |
© 2023 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
Share and Cite
Pang, Y.; Zhou, S.; Zumbo, P.; Betel, D.; Cisse, B. TCF12 Deficiency Impairs the Proliferation of Glioblastoma Tumor Cells and Improves Survival. Cancers 2023, 15, 2033. https://doi.org/10.3390/cancers15072033
Pang Y, Zhou S, Zumbo P, Betel D, Cisse B. TCF12 Deficiency Impairs the Proliferation of Glioblastoma Tumor Cells and Improves Survival. Cancers. 2023; 15(7):2033. https://doi.org/10.3390/cancers15072033
Chicago/Turabian StylePang, Yunong, Sichang Zhou, Paul Zumbo, Doron Betel, and Babacar Cisse. 2023. "TCF12 Deficiency Impairs the Proliferation of Glioblastoma Tumor Cells and Improves Survival" Cancers 15, no. 7: 2033. https://doi.org/10.3390/cancers15072033