103 results on '"Westendorp B"'
Search Results
2. Indian Hedgehog Suppresses a Stromal Cell–Driven Intestinal Immune Response
- Author
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Westendorp, B. Florien, Büller, Nikè V.J.A., Karpus, Olga N., van Dop, Willemijn A., Koster, Jan, Versteeg, Rogier, Koelink, Pim J., Snel, Clinton Y., Meisner, Sander, Roelofs, Joris J.T.H., Uhmann, Anja, Ver Loren van Themaat, Emiel, Heijmans, Jarom, Hahn, Heidi, Muncan, Vanesa, Wildenberg, Manon E., and van den Brink, Gijs R.
- Published
- 2018
- Full Text
- View/download PDF
3. Supplementary Figure legends from Heterozygosity of Chaperone Grp78 Reduces Intestinal Stem Cell Regeneration Potential and Protects against Adenoma Formation
- Author
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van Lidth de Jeude, Jooske F., primary, Spaan, Claudia N., primary, Meijer, Bartolomeus J., primary, Smit, Wouter L., primary, Soeratram, Tanya T.D., primary, Wielenga, Mattheus C.B., primary, Westendorp, B. Florien, primary, Lee, Amy S., primary, Meisner, Sander, primary, Vermeulen, Jacqueline L.M., primary, Wildenberg, Manon E., primary, van den Brink, Gijs R., primary, Muncan, Vanesa, primary, and Heijmans, Jarom, primary
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- 2023
- Full Text
- View/download PDF
4. Figure S2 from Heterozygosity of Chaperone Grp78 Reduces Intestinal Stem Cell Regeneration Potential and Protects against Adenoma Formation
- Author
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van Lidth de Jeude, Jooske F., primary, Spaan, Claudia N., primary, Meijer, Bartolomeus J., primary, Smit, Wouter L., primary, Soeratram, Tanya T.D., primary, Wielenga, Mattheus C.B., primary, Westendorp, B. Florien, primary, Lee, Amy S., primary, Meisner, Sander, primary, Vermeulen, Jacqueline L.M., primary, Wildenberg, Manon E., primary, van den Brink, Gijs R., primary, Muncan, Vanesa, primary, and Heijmans, Jarom, primary
- Published
- 2023
- Full Text
- View/download PDF
5. Data from Heterozygosity of Chaperone Grp78 Reduces Intestinal Stem Cell Regeneration Potential and Protects against Adenoma Formation
- Author
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van Lidth de Jeude, Jooske F., primary, Spaan, Claudia N., primary, Meijer, Bartolomeus J., primary, Smit, Wouter L., primary, Soeratram, Tanya T.D., primary, Wielenga, Mattheus C.B., primary, Westendorp, B. Florien, primary, Lee, Amy S., primary, Meisner, Sander, primary, Vermeulen, Jacqueline L.M., primary, Wildenberg, Manon E., primary, van den Brink, Gijs R., primary, Muncan, Vanesa, primary, and Heijmans, Jarom, primary
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- 2023
- Full Text
- View/download PDF
6. Hedgehog Signaling in Gastrointestinal Morphogenesis and Morphostasis
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Florien Westendorp, B., primary, van Dop, Willemijn A., additional, and van den Brink, Gijs R., additional
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- 2018
- Full Text
- View/download PDF
7. Contributors
- Author
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Akiba, Yasutada, primary, Al Alam, Denise, additional, Al-Sadi, Rana, additional, Aziz, Qasim, additional, Battaglioli, Eric J., additional, Bharucha, Adil E., additional, Blumberg, Richard S., additional, Bohin, Natacha, additional, Bornstein, Joel C., additional, Brierley, Stuart M., additional, Brookes, Simon J.H., additional, Castro, Joel, additional, Chang, Eugene B., additional, Chassaing, Benoit, additional, Cheung, Mary, additional, Ciorba, Matthew A., additional, Crowe, Sheila E., additional, Czerwinski, Michael, additional, Danopoulos, Soula, additional, Das, Soumita, additional, Dempsey, Peter J., additional, den Hartog, Gerco, additional, Enomoto, Hideki, additional, Erickson, Andelain, additional, Ernst, Peter B., additional, Farmer, Adam D., additional, Foong, Jaime P.P., additional, Frey, Mark R., additional, Gewirtz, Andrew T., additional, Ghishan, Fayez K., additional, Gribble, Fiona M., additional, Grundy, Luke, additional, Hao, Marlene M., additional, Harrington, Andrea M., additional, Hennig, Grant W., additional, Hu, Hongzhen, additional, Huizinga, Jan D., additional, Iyer, Shankar S., additional, Kaji, Izumi, additional, Kashyap, Purna C., additional, Kaunitz, Jonathan D., additional, Labus, Jennifer S., additional, Lavoie, Brigitte, additional, Liu, Cambrian Y., additional, Ma, Thomas Y., additional, Ma, Xiaoya, additional, Mawe, Gary M., additional, Merchant, Juanita L., additional, Messer, Jeannette S., additional, Miller, Larry, additional, Naliboff, Bruce D., additional, Nelson, Mark T., additional, Newgreen, Donald F., additional, Nighot, Prashant, additional, Passi, Monica, additional, Polk, D. Brent, additional, Pozo, Maria J., additional, Reimann, Frank, additional, Roberts, Geoffrey P., additional, Roland, Bani C., additional, Ruffle, James K., additional, Said, Hyder, additional, Samuelson, Linda C., additional, Schumacher, Michael A., additional, Shah, Yatrik M., additional, Shea-Donohue, Terez, additional, Shroyer, Noah F., additional, Spence, Jason R., additional, Spencer, Nick J., additional, Spohn, Stephanie N., additional, Stamp, Lincon A., additional, Stenson, William F., additional, Takaki, Miyako, additional, Tillisch, Kirsten, additional, Uesaka, Toshihiro, additional, van den Brink, Gijs R., additional, van Dop, Willemijn A., additional, Vegesna, Anil, additional, von Moltke, Jakob, additional, Wald, Arnold, additional, Westendorp, B. Florien, additional, Whitson, Mathew, additional, Wood, Jackie D., additional, Xu, Hua, additional, Yang, Vincent W., additional, Young, Heather M., additional, Young, Vincent B., additional, Abumrad, Nada A., additional, Alrefai, Waddah A., additional, Ammoury, Rana, additional, Anderson, Gregory J., additional, Asano, Shinji, additional, Bamias, Giorgos, additional, Bhutia, Yangzom D., additional, Blondet, Niviann M., additional, Borel, Patrick, additional, Cifarelli, Vincenza, additional, Collins, James F., additional, Cousins, Robert J., additional, Davidson, Nicholas O., additional, Dawson, Paul A., additional, de Lartigue, Guillaume, additional, Desmarchelier, Charles, additional, Dudeja, Pradeep K., additional, Flores, Shireen R.L., additional, Ganapathy, Vadivel, additional, Ghezzi, Chiara, additional, Gill, Ravinder K., additional, Gorelick, Fred S., additional, Grisham, Matthew B., additional, Harrison, Earl H., additional, Hecht, Gail A., additional, Israel, Dawn A., additional, Jamieson, James D., additional, Katona, Bryson W., additional, Kiela, Pawel R., additional, Kopec, Rachel E., additional, Kowdley, Kris V., additional, LaRusso, Nicholas F., additional, Lee, Seong M., additional, Liddle, Rodger A., additional, Liuzzi, Juan P., additional, Loo, Donald D.F., additional, Lynch, John P., additional, Masyuk, Anatoliy I., additional, Masyuk, Tatyana V., additional, Messner, Donald J., additional, Meyer, Mark B., additional, Murray, Karen F., additional, Nexo, Ebba, additional, Okamoto, Curtis T., additional, Ortega, Bernardo, additional, Pandol, Stephen, additional, Peek, Richard M., additional, Pike, J. Wesley, additional, Priyamvada, Shubha, additional, Proctor, Gordon B., additional, Rajendran, Vazhaikkurichi M., additional, Raybould, Helen E., additional, Rivera-Nieves, Jesus, additional, Said, Hamid M., additional, Sakai, Hideki, additional, Saksena, Seema, additional, Sala-Rabanal, Monica, additional, Schulzke, Jörg-Dieter, additional, Seidler, Ursula E., additional, Shaalan, Abeer K., additional, Sheikh, Irshad A., additional, Thiagarajah, Jay R., additional, Verkman, Alan S., additional, Wang, Xiaoyu, additional, Welling, Paul A., additional, Wolkoff, Allan W., additional, and Wright, Ernest M., additional
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- 2018
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8. CD95 signaling in colorectal cancer
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Hoogwater, Frederik J.H., Steller, Ernst J.A., Westendorp, B. Florien, Borel Rinkes, Inne H.M., and Kranenburg, Onno
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- 2012
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9. Oncogenic K-Ras Turns Death Receptors Into Metastasis-Promoting Receptors in Human and Mouse Colorectal Cancer Cells
- Author
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Hoogwater, Frederik J.H., Nijkamp, Maarten W., Smakman, Niels, Steller, Ernst J.A., Emmink, Benjamin L., Westendorp, B. Florien, Raats, Danielle A.E., Sprick, Martin R., Schaefer, Uta, Van Houdt, Winan J., De Bruijn, Menno T., Schackmann, Ron C.J., Derksen, Patrick W.B., Medema, Jan–Paul, Walczak, Henning, Borel Rinkes, Inne H.M., and Kranenburg, Onno
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- 2010
- Full Text
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10. Synergistic functions of E2F7 and E2F8 are critical to suppress stress-induced skin cancer
- Author
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Thurlings, I, Martínez-López, L M, Westendorp, B, Zijp, M, Kuiper, R, Tooten, P, Kent, L N, Leone, G, Vos, H J, Burgering, B, de Bruin, A, LS Pathobiologie, Dep Gezondheidszorg Landbouwhuisdieren, dPB RMSC, LS Pathobiologie, Dep Gezondheidszorg Landbouwhuisdieren, and dPB RMSC
- Subjects
Keratinocytes ,0301 basic medicine ,Cancer Research ,Skin Neoplasms ,GENES ,DNA damage ,DNA-BINDING DOMAINS ,Apoptosis ,PROGRESSION ,Biology ,medicine.disease_cause ,CELL-PROLIFERATION ,Mice ,03 medical and health sciences ,0302 clinical medicine ,E2F7 Transcription Factor ,Genetics ,medicine ,Animals ,Humans ,E2F1 ,TRANSCRIPTION ,E2F ,Molecular Biology ,ATYPICAL E2FS ,Mice, Knockout ,DAMAGE ,CARCINOGENESIS ,Cell cycle ,medicine.disease ,Repressor Proteins ,030104 developmental biology ,medicine.anatomical_structure ,TARGET ,030220 oncology & carcinogenesis ,FAMILY-MEMBER ,Immunology ,Cancer research ,Original Article ,Skin cancer ,biological phenomena, cell phenomena, and immunity ,Keratinocyte ,Carcinogenesis ,E2F Transcription Factors ,DNA Damage - Abstract
E2F transcription factors are important regulators of the cell cycle, and unrestrained activation of E2F-dependent transcription is considered to be an important driver of tumor formation and progression. Although highly expressed in normal skin and skin cancer, the role of the atypical E2Fs, E2F7 and E2F8, in keratinocyte homeostasis, regeneration and tumorigenesis is unknown. Surprisingly, keratinocyte-specific deletion of E2F7 and E2F8 in mice did not interfere with skin development and wound healing. However, the rate for successful isolation and establishment of E2f7/8-deficient primary keratinocyte cultures was much higher than for wild-type keratinocytes. Moreover, E2f7/8-deficient primary keratinocytes proliferate more efficiently under stress conditions, such as low/high confluence or DNA damage. Application of in vivo stress using the DMBA/TPA skin carcinogenesis protocol revealed that combined inactivation of E2f7/8 enhanced tumorigenesis and accelerated malignant progression. Loss of atypical E2Fs resulted in increased expression of E2F target genes, including E2f1. Additional loss of E2f1 did not rescue, but worsened skin tumorigenesis. We show that loss of E2F7/8 triggers apoptosis via induction of E2F1 in response to stress, indicating that the tumor-promoting effect of E2F7/8 inactivation can be partially compensated via E2F1-dependent apoptosis. Importantly, E2F7/8 repressed a large set of E2F target genes that are highly expressed in human patients with skin cancer. Together, our studies demonstrate that atypical E2Fs act as tumor suppressors, most likely via transcriptional repression of cell cycle genes in response to stress.
- Published
- 2017
11. Chapter 4 - Hedgehog Signaling in Gastrointestinal Morphogenesis and Morphostasis
- Author
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Florien Westendorp, B., van Dop, Willemijn A., and van den Brink, Gijs R.
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- 2018
- Full Text
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12. Chk1 and 14‐3‐3 proteins inhibit atypical E2Fs to prevent a permanent cell cycle arrest
- Author
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Yuan, R., Vos, Harmjan, van Es, Robert M, Chen, J., Burgering, B.M.T., Westendorp, B., de Bruin, A., LS Pathobiologie, Afd Pharmacology, dPB RMSC, LS Pathobiologie, Afd Pharmacology, and dPB RMSC
- Subjects
0301 basic medicine ,Cell cycle checkpoint ,RECOMBINATION ,Apoptosis ,PROGRESSION ,medicine.disease_cause ,0302 clinical medicine ,E2F7 Transcription Factor ,Neoplasms ,TRANSCRIPTION ,Phosphorylation ,S-CHECKPOINT ,Promoter Regions, Genetic ,DNAdamage ,General Neuroscience ,Articles ,Cell cycle ,3. Good health ,Cell biology ,030220 oncology & carcinogenesis ,cell cycle ,biological phenomena, cell phenomena, and immunity ,Protein Binding ,DNA Replication ,Cell Survival ,Cdc25 ,14‐3‐3 proteins ,DNA-DAMAGE RESPONSE ,Biology ,Article ,General Biochemistry, Genetics and Molecular Biology ,REPLICATION-STRESS ,03 medical and health sciences ,Cell Line, Tumor ,medicine ,Humans ,BREAST-CANCER ,CHEK1 ,E2F ,Molecular Biology ,REPAIR ,CDC25 ,General Immunology and Microbiology ,checkpoint kinase 1 ,DNA Replication, Repair & Recombination ,14-3-3 proteins ,Promoter ,Cell Cycle Checkpoints ,atypical E2Fs ,Repressor Proteins ,HEK293 Cells ,030104 developmental biology ,Protein Biosynthesis ,Cancer cell ,biology.protein ,DNA damage ,Carcinogenesis ,HeLa Cells - Abstract
The atypical E2Fs, E2F7 and E2F8, act as potent transcriptional repressors of DNA replication genes providing them with the ability to induce a permanent S‐phase arrest and suppress tumorigenesis. Surprisingly in human cancer, transcript levels of atypical E2Fs are frequently elevated in proliferating cancer cells, suggesting that the tumor suppressor functions of atypical E2Fs might be inhibited through unknown post‐translational mechanisms. Here, we show that atypical E2Fs can be directly phosphorylated by checkpoint kinase 1 (Chk1) to prevent a permanent cell cycle arrest. We found that 14‐3‐3 protein isoforms interact with both E2Fs in a Chk1‐dependent manner. Strikingly, Chk1 phosphorylation and 14‐3‐3‐binding did not relocate or degrade atypical E2Fs, but instead, 14‐3‐3 is recruited to E2F7/8 target gene promoters to possibly interfere with transcription. We observed that high levels of 14‐3‐3 strongly correlate with upregulated transcription of atypical E2F target genes in human cancer. Thus, we reveal that Chk1 and 14‐3‐3 proteins cooperate to inactivate the transcriptional repressor functions of atypical E2Fs. This mechanism might be of particular importance to cancer cells, since they are exposed frequently to DNA‐damaging therapeutic reagents.
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- 2018
13. Colonic CD90+ Crypt Fibroblasts Secrete Semaphorins to Support Epithelial Growth
- Author
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Karpus, Olga N., primary, Westendorp, B. Florien, additional, Vermeulen, Jacqueline L.M., additional, Meisner, Sander, additional, Koster, Jan, additional, Muncan, Vanesa, additional, Wildenberg, Manon E., additional, and van den Brink, Gijs R., additional
- Published
- 2019
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14. Heterozygosity of Chaperone Grp78 Reduces Intestinal Stem Cell Regeneration Potential and Protects against Adenoma Formation
- Author
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van Lidth de Jeude, Jooske F., primary, Spaan, Claudia N., additional, Meijer, Bartolomeus J., additional, Smit, Wouter L., additional, Soeratram, Tanya T.D., additional, Wielenga, Mattheus C.B., additional, Westendorp, B. Florien, additional, Lee, Amy S., additional, Meisner, Sander, additional, Vermeulen, Jacqueline L.M., additional, Wildenberg, Manon E., additional, van den Brink, Gijs R., additional, Muncan, Vanesa, additional, and Heijmans, Jarom, additional
- Published
- 2018
- Full Text
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15. Physiological significance of polyploidization in mammalian cells
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Pandit, S.K., Westendorp, B., de Bruin, A., Tissue Repair, Dep Pathobiologie, Tissue Repair, and Dep Pathobiologie
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Genetics ,Physiological significance ,genetic processes ,fungi ,food and beverages ,pathological conditions, signs and symptoms ,Cell Biology ,Disease ,Chronic injury ,Biology ,Phenotype ,Cell size ,Cell biology ,Polyploidy ,Mice ,Biological property ,Hepatocytes ,Animals ,Humans ,Adaptation ,E2F ,Cell Size - Abstract
Programmed polyploidization occurs in all mammalian species during development and aging in selected tissues, but the biological properties of polyploid cells remain obscure. Spontaneous polyploidization arises during stress and has been observed in a variety of pathological conditions, such as cancer and degenerative diseases. A major challenge in the field is to test the predicted functions of polyploidization in vivo. However, recent genetic mouse models with diminished polyploidization phenotypes represent novel, powerful tools to unravel the biological function of polyploidization. Contrary to a longstanding hypothesis, polyploidization appears to not be required for differentiation and has no obvious impact on proliferation. Instead, polyploidization leads to increased cell size and genetic diversity, which could promote better adaptation to chronic injury or stress. We discuss here the consequences of reducing polyploidization in mice and review which stress responses and molecular signals trigger polyploidization during development and disease.
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- 2013
16. Atypical E2Fs inhibit tumor angiogenesis
- Author
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Weijts, B G M W, primary, Westendorp, B, additional, Hien, B T, additional, Martínez-López, L M, additional, Zijp, M, additional, Thurlings, I, additional, Thomas, R E, additional, Schulte-Merker, S, additional, Bakker, W J, additional, and de Bruin, A, additional
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- 2017
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17. TP53 Regulates Transcription of Genes Involved in G1 Cell Cycle Arrest
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de Bruin, A, primary, Westendorp, B, additional, and Di Stefano, L, additional
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- 2017
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18. Stromal Indian Hedgehog Signaling is Required for Adenoma Formation in Mice
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Bxfcller NV, Rosekrans SL, Metcalfe C, Heijmans J, van Dop WA, Fessler E, Jansen M, Ahn C, Vermeulen JL, Florien Westendorp B, Robanus-Maandag EC, Johan Offerhaus G, Medema JP, D'Haens GR, Wildenberg ME, de Sauvage FJ, Muncan V, and van den Brink GR
- Published
- 2014
19. Synergistic functions of E2F7 and E2F8 are critical to suppress stress-induced skin cancer
- Author
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Thurlings, I, primary, Martínez-López, L M, additional, Westendorp, B, additional, Zijp, M, additional, Kuiper, R, additional, Tooten, P, additional, Kent, L N, additional, Leone, G, additional, Vos, H J, additional, Burgering, B, additional, and de Bruin, A, additional
- Published
- 2016
- Full Text
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20. HIF proteins connect the RB-E2F factors to angiogenesis
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Bakker, W.J., Weijts, B.G.M.W., Westendorp, B., de Bruin, A., Tissue Repair, and Dep Pathobiologie
- Published
- 2013
21. The E2F6 repressor activates gene expression in myocardium resulting in dilated cardiomyopathy
- Author
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Westendorp, B., Major, J.L., Nader, M., Salih, M., Leenen, F.H., Tuana, B.S., Tissue Repair, and Dep Pathobiologie
- Abstract
The E2F/Rb pathway regulates cardiac growth and development and holds great potential as a therapeutic target. The E2F6 repressor is a unique E2F member that acts independently of pocket proteins. Forced expression of E2F6 in mouse myocardium induced heart failure and mortality, with severity of symptoms correlating to E2F6 levels. Echocardiography demonstrated a 37% increase (P>0.05) in left ventricular end-diastolic diameter and reduced ejection fraction (0.05) in young transgenic (Tg) mice. Microarray and qPCR analysis revealed a paradoxical increase in E2F-responsive genes, which regulate the cell cycle, without changes in cardiomyocyte cell number or size in Tg mice. Young adult Tg mice displayed a 75% (P>0.01) decrease in gap junction protein connexin-43, resulting in abnormal electrocardiogram including a 24% (P>0.05) increase in PR interval. Further, mir-206, which targets connexin-43, was up-regulated 10-fold (P>0.05) in Tg myocardium. The mitogen-activated protein kinase pathway, which regulates the levels of miR-206 and connexin-43, was activated in Tg hearts. Thus, deregulated E2F6 levels evoked abnormal gene expression at transcriptional and post-transcriptional levels, leading to cardiac remodeling and dilated cardiomyopathy. The data highlight an unprecedented role for the strict regulation of the E2F pathway in normal postnatal cardiac function.—Westendorp, B., Major, J. L., Nader, M., Salih, M., Leenen, F. H. H., Tuana, B. S. The E2F6 repressor activates gene expression in myocardium resulting in dilated cardiomyopathy. FASEB J. 26, 2569-2579 (2012). www.fasebj.org
- Published
- 2012
22. E2F7 and E2F8 promote angiogenesis through transcriptional activation of VEGFA in cooperation with HIF1
- Author
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Weijts, B.G.M.W., Bakker, W.J., Cornelissen, P.W.A., Liang, K., Schaftenaar, F.H., Westendorp, B., de Wolf, C.A.C.M.T., Paciejewska, M., Scheele, C.L.G.J., Kent, L., Leone, G., Schulte-Merker, S., de Bruin, A., and Hubrecht Institute for Developmental Biology and Stem Cell Research
- Subjects
embryonic-development ,proliferation ,growth-factor gene ,zebrafish ,hif-1-alpha ,vascular development ,o-2 homeostasis ,hypoxia-inducible factor-1 ,Experimentele Zoologie ,WIAS ,cancer ,Experimental Zoology ,factor 1-alpha - Abstract
The E2F family of transcription factors plays an important role in controlling cell-cycle progression. While this is their best-known function, we report here novel functions for the newest members of the E2F family, E2F7 and E2F8 (E2F7/8). We show that simultaneous deletion of E2F7/8 in zebrafish and mice leads to severe vascular defects during embryonic development. Using a panel of transgenic zebrafish with fluorescent-labelled blood vessels, we demonstrate that E2F7/8 are essential for proper formation of blood vessels. Despite their classification as transcriptional repressors, we provide evidence for a molecular mechanism through which E2F7/8 activate the transcription of the vascular endothelial growth factor A (VEGFA), a key factor in guiding angiogenesis. We show that E2F7/8 directly bind and stimulate the VEGFA promoter independent of canonical E2F binding elements. Instead, E2F7/8 form a transcriptional complex with the hypoxia inducible factor 1 (HIF1) to stimulate VEGFA promoter activity. These results uncover an unexpected link between E2F7/8 and the HIF1-VEGFA pathway providing a molecular mechanism by which E2F7/8 control angiogenesis.
- Published
- 2012
23. E2F7 represses a network of oscillating cell cycle genes to control S-phase progression
- Author
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Westendorp, B., Mokry, M., Groot Koerkamp, M.J., Holstege, F.C.P., Cuppen, E.P.J.G., de Bruin, A., Tissue Repair, Dep Pathobiologie, Tissue Repair, Dep Pathobiologie, and Hubrecht Institute for Developmental Biology and Stem Cell Research
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Periodicity ,Transcription, Genetic ,DNA repair ,Biology ,S Phase ,Mice ,E2F7 Transcription Factor ,Transcription (biology) ,Consensus Sequence ,Genetics ,Animals ,Gene Regulatory Networks ,E2F ,Gene ,Binding Sites ,Base Sequence ,DNA replication ,G1 Phase ,Promoter ,Genomics ,Cell Cycle Gene ,Molecular biology ,Genes, cdc ,Repressor Proteins ,Gene Expression Regulation ,S Phase Cell Cycle Checkpoints ,E2F Transcription Factors ,DNA Damage - Abstract
E2F transcription factors are known to be important for timely activation of G 1 /S and G 2 /M genes required for cell cycle progression, but transcriptional mechanisms for deactivation of cell cycle-regulated genes are unknown. Here, we show that E2F7 is highly expressed during mid to late S-phase, occupies promoters of G 1 /S-regulated genes and represses their transcription. ChIP-seq analysis revealed that E2F7 binds preferentially to genomic sites containing the TTCCCGCC motif, which closely resembles the E2F consensus site. We identified 89 target genes that carry E2F7 binding sites close to the transcriptional start site and that are directly repressed by short-term induction of E2F7. Most of these target genes are known to be activated by E2Fs and are involved in DNA replication, metabolism and DNA repair. Importantly, induction of E2F7 during G 0 -G 1 /S resulted in S-phase arrest and DNA damage, whereas expression of E2F7 during G 2 /M failed to disturb cell cycle progression. These findings provide strong evidence that E2F7 directly controls the downswing of oscillating G 1 /S genes during S-phase progression.
- Published
- 2012
24. Stromal Indian Hedgehog Signaling Is Required for Intestinal Adenoma Formation in Mice
- Author
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Büller, Nikè V.J.A., primary, Rosekrans, Sanne L., additional, Metcalfe, Ciara, additional, Heijmans, Jarom, additional, van Dop, Willemijn A., additional, Fessler, Evelyn, additional, Jansen, Marnix, additional, Ahn, Christina, additional, Vermeulen, Jacqueline L.M., additional, Westendorp, B. Florien, additional, Robanus-Maandag, Els C., additional, Offerhaus, G. Johan, additional, Medema, Jan Paul, additional, D’Haens, Geert R.A.M., additional, Wildenberg, Manon E., additional, de Sauvage, Frederic J., additional, Muncan, Vanesa, additional, and van den Brink, Gijs R., additional
- Published
- 2015
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25. Atypical E2Fs inhibit tumor angiogenesis
- Author
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Weijts, B G M W, Westendorp, B, Hien, B T, Martínez-López, L M, Zijp, M, Thurlings, I, Thomas, R E, Schulte-Merker, S, Bakker, W J, and de Bruin, A
- Abstract
Atypical E2F transcription factors (E2F7 and E2F8) function as key regulators of cell cycle progression and their inactivation leads to spontaneous cancer formation in mice. However, the mechanism of the tumor suppressor functions of E2F7/8 remain obscure. In this study we discovered that atypical E2Fs control tumor angiogenesis, one of the hallmarks of cancer. We genetically inactivated atypical E2Fs in epithelial and mesenchymal neoplasm and analyzed blood vessel formation in three different animal models of cancer. Tumor formation was either induced by application of 7,12-Dimethylbenz(a)anthracene/12-O-Tetradecanoylphorbol-13-acetate or by Myc/Ras overexpression. To our surprise, atypical E2Fs suppressed tumor angiogenesis in all three cancer models, which is in a sharp contrast to previous findings showing that atypical E2Fs promote angiogenesis during fetal development in mice and zebrafish. Real-time imaging in zebrafish displayed that fluorescent-labeled blood vessels showed enhanced intratumoral branching in xenografted E2f7/8-deficient neoplasms compared with E2f7/8-proficient neoplasms. DLL4 expression, a key negative inhibitor of vascular branching, was decreased in E2f7/8-deficient neoplastic cells, indicating that E2F7/8 might inhibit intratumoral vessel branching via induction of DLL4.
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- 2018
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26. CD95 is a key mediator of invasion and accelerated outgrowth of mouse colorectal liver metastases following radiofrequency ablation
- Author
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Nijkamp, Maarten W., primary, Hoogwater, Frederik J.H., additional, Steller, Ernst J.A., additional, Westendorp, B. Florien, additional, van der Meulen, Taco A., additional, Leenders, Martijn W.H., additional, Borel Rinkes, Inne H.M., additional, and Kranenburg, Onno, additional
- Published
- 2010
- Full Text
- View/download PDF
27. 263 Does LXR-alpha connect cholesterol to the renin angiotensin system?
- Author
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DEBOER, R, primary, KUIPERS, I, additional, VANGENNE, L, additional, VANDERHARST, P, additional, WESTENDORP, B, additional, VANVELDHUISEN, D, additional, and VANGILST, W, additional
- Published
- 2006
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28. 65 Adverse renal effects of combined hydrochlorothiazide and ACE inhibitor therapy in rats with myocardial infarction
- Author
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WESTENDORP, B, primary, HAMMING, I, additional, NAVIS, G, additional, VANGOOR, H, additional, BUIKEMA, H, additional, VANGILST, W, additional, and SCHOEMAKER, R, additional
- Published
- 2006
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- View/download PDF
29. Adverse renal effects of hydrochlorothiazide in rats with myocardial infarction treated with ACE inhibition
- Author
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Westendorp, B, Hamming, [No Value], Navis, G, van Goor, H, Buikema, H, van Gilst, W, Groningen University Institute for Drug Exploration (GUIDE), Cardiovascular Centre (CVC), Groningen Institute for Organ Transplantation (GIOT), Groningen Kidney Center (GKC), Lifestyle Medicine (LM), and Vascular Ageing Programme (VAP)
30. 263 Does LXR-alpha connect cholesterol to the renin angiotensin system?
- Author
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De Boer, R.A., Kuipers, I., van Genne, L., van der Harst, P., Westendorp, B., van Veldhuisen, D.J., and van Gilst, W.H.
- Subjects
CHOLESTEROL ,ANGIOTENSINS - Abstract
An abstract of the study "Does LXR-alpha connect cholesterol to the renin angiotensin system?," by R. A. De Boer and colleagues is presented.
- Published
- 2006
31. Discovery of Tissue-Specific Functions of Atypical E2Fs in Cancer
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Eva Moreno Iglesias, Bruin, A. de, Westendorp, B., and University Utrecht
- Subjects
Genetically modified mouse ,stomatognathic diseases ,business.industry ,medicine ,Cancer research ,Tissue specific ,Cancer ,biological phenomena, cell phenomena, and immunity ,Liver cancer ,medicine.disease ,business ,atypical E2Fs ,RB ,transgenic mice ,Polyploidization ,liver cancer ,NAFLD - Abstract
The cell cycle control system ensures that the genome is properly duplicated and evenly distributed over the daughter cells. This system is highly conserved and regulated by a plethora of genes. Among this regulatory system is the Rb-E2F network, which controls the expression of key cell cycle genes. Thus, it is not surprising that Rb is often lost or mutated in cancer, resulting in hyperactivated E2F transcription and enhanced tumorigenesis. To counteract enhanced E2F transcription, alternative mechanisms that inhibit E2F transcription to efficiently block proliferation of cancer cells are needed. In this thesis, we manipulated E2F-dependent transcription by altering the expression of E2F7 and E2F8 in mouse models. E2F7/8 are atypical cell cycle inhibitors belonging to the E2F family that possess Rb-independent repressive function. We hypothesized that overactivation of atypical E2F repressors efficiently blocks cancer cell proliferation. The studies from this thesis demonstrated that downregulation of E2F transcription by boosting E2F7/8 expression impaired the ability of liver cancer cells to progress through the cell cycle and suppressed tumor growth. In addition, unrestrained E2F dependent transcription due to loss of atypical E2Fs and RB, induced tumorigenesis in a tissue cell-type specific manner via a mechanism that involves maintenance of genomic instability. Lastly, we unravel novel functions of atypical E2Fs in lipid metabolism by controlling liver polyploidization. Taken together, this thesis has contributed to our understanding of the role of atypical E2Fs and Rb in cancer and provides scientific evidence that inhibition of E2F-depedent transcription could be a novel therapeutic option in oncology .
- Published
- 2021
32. A reverse brake for the cell cycle.
- Author
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Westendorp B
- Subjects
- Animals, Humans, Gene Duplication, S Phase, Endoreduplication
- Abstract
Mitogenic signaling acts beyond S-phase entry to prevent whole-genome duplications.
- Published
- 2024
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33. Assessment of Kinome-Wide Activity Remodeling upon Picornavirus Infection.
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Veth TS, Nouwen LV, Zwaagstra M, Lyoo H, Wierenga KA, Westendorp B, Altelaar MAFM, Berkers C, van Kuppeveld FJM, and Heck AJR
- Subjects
- Humans, HeLa Cells, Proteome metabolism, Protein Kinases metabolism, Virus Replication, Phosphorylation, Picornaviridae physiology, Picornaviridae enzymology, Picornaviridae Infections virology, Picornaviridae Infections metabolism
- Abstract
Picornaviridae represent a large family of single-stranded positive RNA viruses of which different members can infect both humans and animals. These include the enteroviruses (e.g., poliovirus, coxsackievirus, and rhinoviruses) as well as the cardioviruses (e.g., encephalomyocarditis virus). Picornaviruses have evolved to interact with, use, and/or evade cellular host systems to create the optimal environment for replication and spreading. It is known that viruses modify kinase activity during infection, but a proteome-wide overview of the (de)regulation of cellular kinases during picornavirus infection is lacking. To study the kinase activity landscape during picornavirus infection, we here applied dedicated targeted mass spectrometry-based assays covering ∼40% of the human kinome. Our data show that upon infection, kinases of the MAPK pathways become activated (e.g., ERK1/2, RSK1/2, JNK1/2/3, and p38), while kinases involved in regulating the cell cycle (e.g., CDK1/2, GWL, and DYRK3) become inactivated. Additionally, we observed the activation of CHK2, an important kinase involved in the DNA damage response. Using pharmacological kinase inhibitors, we demonstrate that several of these activated kinases are essential for the replication of encephalomyocarditis virus. Altogether, the data provide a quantitative understanding of the regulation of kinome activity induced by picornavirus infection, providing a resource important for developing novel antiviral therapeutic interventions., Competing Interests: Conflict of interest The authors declare no competing interests., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)
- Published
- 2024
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34. MUC13 negatively regulates tight junction proteins and intestinal epithelial barrier integrity via protein kinase C.
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Segui-Perez C, Stapels DAC, Ma Z, Su J, Passchier E, Westendorp B, Wubbolts RW, Wu W, van Putten JPM, and Strijbis K
- Subjects
- Humans, Intestines, Intestinal Mucosa metabolism, Tight Junctions metabolism, Occludin, Mucins metabolism, Epithelial Cells metabolism, Tight Junction Proteins metabolism, Protein Kinase C metabolism
- Abstract
Glycosylated mucin proteins contribute to the essential barrier function of the intestinal epithelium. The transmembrane mucin MUC13 is an abundant intestinal glycoprotein with important functions for mucosal maintenance that are not yet completely understood. We demonstrate that in human intestinal epithelial monolayers, MUC13 localized to both the apical surface and the tight junction (TJ) region on the lateral membrane. MUC13 deletion resulted in increased transepithelial resistance (TEER) and reduced translocation of small solutes. TEER buildup in ΔMUC13 cells could be prevented by addition of MLCK, ROCK or protein kinase C (PKC) inhibitors. The levels of TJ proteins including claudins and occludin were highly increased in membrane fractions of MUC13 knockout cells. Removal of the MUC13 cytoplasmic tail (CT) also altered TJ composition but did not affect TEER. The increased buildup of TJ complexes in ΔMUC13 and MUC13-ΔCT cells was dependent on PKC. The responsible PKC member might be PKCδ (or PRKCD) based on elevated protein levels in the absence of full-length MUC13. Our results demonstrate for the first time that a mucin protein can negatively regulate TJ function and stimulate intestinal barrier permeability., Competing Interests: Competing interests The authors declare no competing or financial interests., (© 2024. Published by The Company of Biologists Ltd.)
- Published
- 2024
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35. Editorial: The role of E2F transcription factors in cancer.
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Iglesias-Ara A and Westendorp B
- Abstract
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.
- Published
- 2024
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36. Glycosylated extracellular mucin domains protect against SARS-CoV-2 infection at the respiratory surface.
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Chatterjee M, Huang LZX, Mykytyn AZ, Wang C, Lamers MM, Westendorp B, Wubbolts RW, van Putten JPM, Bosch BJ, Haagmans BL, and Strijbis K
- Subjects
- Humans, Angiotensin-Converting Enzyme 2, SARS-CoV-2 metabolism, CA-125 Antigen metabolism, Lung metabolism, Polysaccharides, Mucins metabolism, COVID-19
- Abstract
Mucins play an essential role in protecting the respiratory tract against microbial infections while also acting as binding sites for bacterial and viral adhesins. The heavily O-glycosylated gel-forming mucins MUC5AC and MUC5B eliminate pathogens by mucociliary clearance. Transmembrane mucins MUC1, MUC4, and MUC16 can restrict microbial invasion at the apical surface of the epithelium. In this study, we determined the impact of host mucins and mucin glycans on epithelial entry of SARS-CoV-2. Human lung epithelial Calu-3 cells express the SARS-CoV-2 entry receptor ACE2 and high levels of glycosylated MUC1, but not MUC4 and MUC16, on their cell surface. The O-glycan-specific mucinase StcE specifically removed the glycosylated part of the MUC1 extracellular domain while leaving the underlying SEA domain and cytoplasmic tail intact. StcE treatment of Calu-3 cells significantly enhanced infection with SARS-CoV-2 pseudovirus and authentic virus, while removal of terminal mucin glycans sialic acid and fucose from the epithelial surface did not impact viral entry. In Calu-3 cells, the transmembrane mucin MUC1 and ACE2 are located to the apical surface in close proximity and StcE treatment results in enhanced binding of purified spike protein. Both MUC1 and MUC16 are expressed on the surface of human organoid-derived air-liquid interface (ALI) differentiated airway cultures and StcE treatment led to mucin removal and increased levels of SARS-CoV-2 replication. In these cultures, MUC1 was highly expressed in non-ciliated cells while MUC16 was enriched in goblet cells. In conclusion, the glycosylated extracellular domains of different transmembrane mucins might have similar protective functions in different respiratory cell types by restricting SARS-CoV-2 binding and entry., Competing Interests: The authors declare no competing interests., (Copyright: © 2023 Chatterjee et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)
- Published
- 2023
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37. Single cell analysis of docosahexaenoic acid suppression of sequential LPS-induced proinflammatory and interferon-regulated gene expression in the macrophage.
- Author
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Wierenga KA, Riemers FM, Westendorp B, Harkema JR, and Pestka JJ
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- Mice, Animals, Lipopolysaccharides pharmacology, Interferons metabolism, NF-kappa B metabolism, Single-Cell Analysis, Toll-Like Receptor 4 metabolism, Macrophages, Cytokines metabolism, Gene Expression, Docosahexaenoic Acids pharmacology, Docosahexaenoic Acids metabolism, Fatty Acids, Omega-3 metabolism
- Abstract
Preclinical and clinical studies suggest that consumption of long chain omega-3 polyunsaturated fatty acids (PUFAs) reduces severity of chronic inflammatory and autoimmune diseases. While these ameliorative effects are conventionally associated with downregulated expression of proinflammatory cytokine and chemokine genes, our laboratory has recently identified Type 1 interferon (IFN1)-regulated gene expression to be another key target of omega-3 PUFAs. Here we used single cell RNA sequencing (scRNAseq) to gain new mechanistic perspectives on how the omega-3 PUFA docosahexaenoic acid (DHA) influences TLR4-driven proinflammatory and IFN1-regulated gene expression in a novel self-renewing murine fetal liver-derived macrophage (FLM) model. FLMs were cultured with 25 µM DHA or vehicle for 24 h, treated with modest concentration of LPS (20 ng/ml) for 1 and 4 h, and then subjected to scRNAseq using the 10X Chromium System. At 0 h (i.e., in the absence of LPS), DHA increased expression of genes associated with the NRF2 antioxidant response (e.g. Sqstm1 , Hmox1, Chchd10 ) and metal homeostasis (e.g. Mt1 , Mt2, Ftl1, Fth1 ), both of which are consistent with DHA-induced polarization of FLMs to a more anti-inflammatory phenotype. At 1 h post-LPS treatment, DHA inhibited LPS-induced cholesterol synthesis genes (e.g. Scd1, Scd2 , Pmvk, Cyp51, Hmgcs1 , and Fdps) which potentially could contribute to interference with TLR4-mediated inflammatory signaling. At 4 h post-LPS treatment, LPS-treated FLMs reflected a more robust inflammatory response including upregulation of proinflammatory cytokine (e.g. Il1a, Il1b, Tnf ) and chemokine (e.g. Ccl2, Ccl3, Ccl4, Ccl7 ) genes as well as IFN1-regulated genes (e.g. Irf7, Mx1, Oasl1, Ifit1 ), many of which were suppressed by DHA. Using single-cell regulatory network inference and clustering (SCENIC) to identify gene expression networks, we found DHA modestly downregulated LPS-induced expression of NF-κB-target genes. Importantly, LPS induced a subset of FLMs simultaneously expressing NF-κB- and IRF7/STAT1/STAT2-target genes that were conspicuously absent in DHA-pretreated FLMs. Thus, DHA potently targeted both the NF-κB and the IFN1 responses. Altogether, scRNAseq generated a valuable dataset that provides new insights into multiple overlapping mechanisms by which DHA may transcriptionally or post-transcriptionally regulate LPS-induced proinflammatory and IFN1-driven responses in macrophages., 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 © 2022 Wierenga, Riemers, Westendorp, Harkema and Pestka.)
- Published
- 2022
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38. Inhibition of polyploidization in Pten-deficient livers reduces steatosis.
- Author
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Moreno E, Matondo AB, Bongiovanni L, van de Lest CHA, Molenaar MR, Toussaint MJM, van Essen SC, Houweling M, Helms JB, Westendorp B, and de Bruin A
- Subjects
- Animals, Hepatocytes metabolism, Lipids, Liver pathology, Mice, PTEN Phosphohydrolase genetics, PTEN Phosphohydrolase metabolism, Peroxisome Proliferator-Activated Receptors metabolism, Phosphatidylinositol 3-Kinases, Proto-Oncogene Proteins c-akt, Fatty Liver pathology, Liver Neoplasms pathology
- Abstract
The tumour suppressor PTEN is a negative regulator of the PI3K/AKT signalling pathway. Liver-specific deletion of Pten in mice results in the hyper-activation PI3K/AKT signalling accompanied by enhanced genome duplication (polyploidization), marked lipid accumulation (steatosis) and formation of hepatocellular carcinomas. However, it is unknown whether polyploidization in this model has an impact on the development of steatosis and the progression towards liver cancer. Here, we used a liver-specific conditional knockout approach to delete Pten in combination with deletion of E2f7/8, known key inducers of polyploidization. As expected, Pten deletion caused severe steatosis and liver tumours accompanied by enhanced polyploidization. Additional deletion of E2f7/8 inhibited polyploidization, alleviated Pten-induced steatosis without affecting lipid species composition and accelerated liver tumour progression. Global transcriptomic analysis showed that inhibition of polyploidization in Pten-deficient livers resulted in reduced expression of genes involved in energy metabolism, including PPAR-gamma signalling. However, we find no evidence that deregulated genes in Pten-deficient livers are direct transcriptional targets of E2F7/8, supporting that reduction in steatosis and progression towards liver cancer are likely consequences of inhibiting polyploidization. Lastly, flow cytometry and image analysis on isolated primary wildtype mouse hepatocytes provided further support that polyploid cells can accumulate more lipid droplets than diploid hepatocytes. Collectively, we show that polyploidization promotes steatosis and function as an important barrier against liver tumour progression in Pten-deficient livers., (© 2022 The Authors. Liver International published by John Wiley & Sons Ltd.)
- Published
- 2022
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39. Mechanisms used by cancer cells to tolerate drug-induced replication stress.
- Author
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Segeren HA and Westendorp B
- Subjects
- Ataxia Telangiectasia Mutated Proteins metabolism, Checkpoint Kinase 1 metabolism, DNA Damage, Humans, NF-E2-Related Factor 2, S Phase Cell Cycle Checkpoints, Stress, Physiological, DNA Replication, Neoplasms drug therapy, Neoplasms genetics
- Abstract
Activation of oncogenes in cancer cells forces cell proliferation, leading to DNA replication stress (RS). As a consequence, cancer cells heavily rely on the intra S-phase checkpoint for survival. This fundamental principle formed the basis for the development of inhibitors against key players of the intra S-phase checkpoint, ATR and CHK1. These drugs are often combined with chemotherapeutic drugs that interfere with DNA replication to exacerbate RS and exhaust the intra S-phase checkpoint in cancer cells. However, drug resistance impedes efficient clinical use, suggesting that some cancer cells tolerate severe RS. In this review, we describe how an increased nucleotide pool, boosted stabilization and repair of stalled forks and firing of dormant origins fortify the RS response in cancer cells. Notably, the vast majority of the genes that confer RS tolerance are regulated by the E2F and NRF2 transcription factors. These transcriptional programs are frequently activated in cancer cells, allowing simultaneous activation of multiple tolerance avenues. We propose that the E2F and NRF2 transcriptional programs can be used as biomarker to select patients for treatment with RS-inducing drugs and as novel targets to kill RS-tolerant cancer cells. Together, this review aims to provide a framework to maximally exploit RS as an Achilles' heel of cancer cells., (Copyright © 2022 The Authors. Published by Elsevier B.V. All rights reserved.)
- Published
- 2022
- Full Text
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40. CDC6: A novel canine tumour biomarker detected in circulating extracellular vesicles.
- Author
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Andriessen A, Bongiovanni L, Driedonks TAP, van Liere E, Seijger A, Hegeman CV, van Nimwegen SA, Galac S, Westendorp B, Nolte-'t Hoen ENM, and de Bruin A
- Subjects
- Animals, Biomarkers, Tumor metabolism, Dogs, Liquid Biopsy methods, Liquid Biopsy veterinary, RNA, Messenger genetics, RNA, Messenger metabolism, Dog Diseases diagnosis, Dog Diseases metabolism, Extracellular Vesicles, Neoplasms diagnosis, Neoplasms metabolism, Neoplasms veterinary
- Abstract
Circulating nucleic acids and extracellular vesicles (EV) represent novel biomarkers to diagnose cancer. The non-invasive nature of these so-called liquid biopsies provides an attractive alternative to tissue biopsy-based cancer diagnostics. This study aimed to investigate if circulating cell cycle-related E2F target transcripts can be used to diagnose tumours in canine tumour patients with different types of tumours. Furthermore, we assessed if these mRNAs are localised within circulating EV. We isolated total RNA from the plasma of 20 canine tumour patients and 20 healthy controls. Four E2F target genes (CDC6, DHFR, H2AFZ and ATAD2) were selected based on the analysis of published data of tumour samples available in public databases. We performed reverse transcription and quantitative real-time PCR to analyse the plasma levels of selected E2F target transcripts. All four E2F target transcripts were detectable in the plasma of canine tumour patients. CDC6 mRNA levels were significantly higher in the plasma of canine tumour patients compared to healthy controls. A subset of canine tumour patient and healthy control plasma samples (n = 7) were subjected to size exclusion chromatography in order to validate association of the E2F target transcripts to circulating EV. For CDC6, EV analysis enhanced their detectability compared to total plasma analysis. In conclusion, our study reveals circulating CDC6 as a promising non-invasive biomarker to diagnose canine tumours., (© 2021 The Authors. Veterinary and Comparative Oncology published by John Wiley & Sons Ltd.)
- Published
- 2022
- Full Text
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41. Oncogenic RAS sensitizes cells to drug-induced replication stress via transcriptional silencing of P53.
- Author
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Segeren HA, van Liere EA, Riemers FM, de Bruin A, and Westendorp B
- Subjects
- Ataxia Telangiectasia Mutated Proteins genetics, Checkpoint Kinase 1 genetics, DNA Damage, DNA Replication genetics, Humans, Protein Kinase Inhibitors pharmacology, Signal Transduction, Genes, ras, Tumor Suppressor Protein p53 genetics
- Abstract
Cancer cells often experience high basal levels of DNA replication stress (RS), for example due to hyperactivation of oncoproteins like MYC or RAS. Therefore, cancer cells are considered to be sensitive to drugs that exacerbate the level of RS or block the intra S-phase checkpoint. Consequently, RS-inducing drugs including ATR and CHK1 inhibitors are used or evaluated as anti-cancer therapies. However, drug resistance and lack of biomarkers predicting therapeutic efficacy limit efficient use. This raises the question what determines sensitivity of individual cancer cells to RS. Here, we report that oncogenic RAS does not only enhance the sensitivity to ATR/CHK1 inhibitors by directly causing RS. Instead, we observed that HRAS
G12V dampens the activation of the P53-dependent transcriptional response to drug-induced RS, which in turn confers sensitivity to RS. We demonstrate that inducible expression of HRASG12V sensitized cells to ATR and CHK1 inhibitors. Using RNA-sequencing of FACS-sorted cells we discovered that P53 signaling is the sole transcriptional response to RS. However, oncogenic RAS attenuates the transcription of P53 and TGF-β pathway components which consequently dampens P53 target gene expression. Accordingly, live cell imaging showed that HRASG12V exacerbates RS in S/G2-phase, which could be rescued by stabilization of P53. Thus, our results demonstrate that transcriptional control of P53 target genes is the prime determinant in the response to ATR/CHK1 inhibitors and show that hyperactivation of the MAPK pathway impedes this response. Our findings suggest that the level of oncogenic MAPK signaling could predict sensitivity to intra-S-phase checkpoint inhibition in cancers with intact P53., (© 2022. The Author(s).)- Published
- 2022
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42. Regulation of a progenitor gene program by SOX4 is essential for mammary tumor proliferation.
- Author
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Roukens MG, Frederiks CL, Seinstra D, Braccioli L, Khalil AA, Pals C, De Neck S, Bornes L, Beerling E, Mokry M, de Bruin A, Westendorp B, van Rheenen J, and Coffer PJ
- Subjects
- Animals, Breast Neoplasms genetics, CRISPR-Cas Systems, Cell Cycle Proteins genetics, Epithelial-Mesenchymal Transition, Female, Gene Expression Regulation, Neoplastic, Gene Silencing, Humans, Lung Neoplasms genetics, Mice, Neoplasm Transplantation, Organoids pathology, Breast Neoplasms pathology, Lung Neoplasms pathology, Lung Neoplasms secondary, Organoids transplantation, SOXC Transcription Factors genetics
- Abstract
In breast cancer the transcription factor SOX4 has been shown to be associated with poor survival, increased tumor size and metastasis formation. This has mostly been attributed to the ability of SOX4 to regulate Epithelial-to-Mesenchymal-Transition (EMT). However, SOX4 regulates target gene transcription in a context-dependent manner that is determined by the cellular and epigenetic state. In this study we have investigated the loss of SOX4 in mammary tumor development utilizing organoids derived from a PyMT genetic mouse model of breast cancer. Using CRISPR/Cas9 to abrogate SOX4 expression, we found that SOX4 is required for inhibiting differentiation by regulating a subset of genes that are highly activated in fetal mammary stem cells (fMaSC). In this way, SOX4 re-activates an oncogenic transcriptional program that is regulated in many progenitor cell-types during embryonic development. SOX4-knockout organoids are characterized by the presence of more differentiated cells that exhibit luminal or basal gene expression patterns, but lower expression of cell cycle genes. In agreement, primary tumor growth and metastatic outgrowth in the lungs are impaired in SOX4
KO tumors. Finally, SOX4KO tumors show a severe loss in competitive capacity to grow out compared to SOX4-proficient cells in primary tumors. Our study identifies a novel role for SOX4 in maintaining mammary tumors in an undifferentiated and proliferative state. Therapeutic manipulation of SOX4 function could provide a novel strategy for cancer differentiation therapy, which would promote differentiation and inhibit cycling of tumor cells., (© 2021. The Author(s).)- Published
- 2021
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43. Collection of cells for single-cell RNA sequencing using high-resolution fluorescence microscopy.
- Author
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Segeren HA, Andree KC, Oomens L, and Westendorp B
- Subjects
- Base Sequence genetics, Flow Cytometry methods, Gene Expression Profiling methods, High-Throughput Nucleotide Sequencing methods, Humans, RNA genetics, Sequence Analysis, RNA methods, Transcriptome genetics, Exome Sequencing methods, Workflow, Cell Separation methods, Microscopy, Fluorescence methods, Single-Cell Analysis methods
- Abstract
FACS sorting followed by single-cell RNA-sequencing (SORT-Seq) is a popular procedure to select cells of interest for single-cell transcriptomics. However, FACS is not suitable for measurement of subcellular distribution of fluorescence or for small samples (<1,000 cells). The VYCAP puncher system overcomes these limitations. Here, we describe a workflow to capture, image, and collect fluorescent human retina pigment epithelium cells for SORT-Seq using this system. The workflow can be used for any cell type with a diameter of ∼5-50 μm. For complete details on the use and execution of this protocol, please refer to Segeren et al. (2020)., Competing Interests: H.A.S. and B.W. declare no competing interests. L.O. and K.C.A. are employed by VYCAP B.V., (© 2021 The Author(s).)
- Published
- 2021
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44. Atypical E2Fs either Counteract or Cooperate with RB during Tumorigenesis Depending on Tissue Context.
- Author
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Moreno E, Pandit SK, Toussaint MJM, Bongiovanni L, Harkema L, van Essen SC, van Liere EA, Westendorp B, and de Bruin A
- Abstract
E2F-transcription factors activate many genes involved in cell cycle progression, DNA repair, and apoptosis. Hence, E2F-dependent transcription must be tightly regulated to prevent tumorigenesis, and therefore metazoan cells possess multiple E2F regulation mechanisms. The best-known is the Retinoblastoma protein (RB), which is mutated in many cancers. Atypical E2Fs (E2F7 and -8) can repress E2F-target gene expression independently of RB and are rarely mutated in cancer. Therefore, they may act as emergency brakes in RB-mutated cells to suppress tumor growth. Currently, it is unknown if and how RB and atypical E2Fs functionally interact in vivo . Here, we demonstrate that mice with liver-specific combinatorial deletion of Rb and E2f7/8 have reduced life-spans compared to E2f7/8 or Rb deletion alone. This was associated with increased proliferation and enhanced malignant progression of liver tumors. Hence, atypical repressor E2Fs and RB cooperatively act as tumor suppressors in hepatocytes. In contrast, loss of either E2f7 or E2f8 largely prevented the formation of pituitary tumors in Rb
+/- mice. To test whether atypical E2Fs can also function as oncogenes independent of RB loss, we induced long-term overexpression of E2f7 or E2f8 in mice. E2F7 and -8 overexpression increased the incidence of tumors in the lungs, but not in other tissues. Collectively, these data show that atypical E2Fs can promote but also inhibit tumorigenesis depending on tissue type and RB status. We propose that the complex interactions between atypical E2Fs and RB on maintenance of genetic stability underlie this context-dependency.- Published
- 2021
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45. E2F7 Is a Potent Inhibitor of Liver Tumor Growth in Adult Mice.
- Author
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Moreno E, Toussaint MJM, van Essen SC, Bongiovanni L, van Liere EA, Koster MH, Yuan R, van Deursen JM, Westendorp B, and de Bruin A
- Subjects
- Animals, Apoptosis physiology, Cell Cycle physiology, DNA Damage, E2F7 Transcription Factor deficiency, E2F7 Transcription Factor genetics, HeLa Cells, Humans, Liver Neoplasms genetics, Male, Mice, Mice, Inbred C57BL, Mice, Knockout, Repressor Proteins deficiency, Repressor Proteins genetics, Transcriptional Activation, Cell Proliferation, E2F7 Transcription Factor physiology, Hepatocytes metabolism, Liver Neoplasms pathology, Repressor Proteins physiology
- Abstract
Background and Aims: Up-regulation of the E2F-dependent transcriptional network has been identified in nearly every human malignancy and is an important driver of tumorigenesis. Two members of the E2F family, E2F7 and E2F8, are potent repressors of E2F-dependent transcription. They are atypical in that they do not bind to dimerization partner proteins and are not controlled by retinoblastoma protein. The physiological relevance of E2F7 and E2F8 remains incompletely understood, largely because tools to manipulate their activity in vivo have been lacking., Approach and Results: Here, we generated transgenic mice with doxycycline-controlled transcriptional activation of E2f7 and E2f8 and induced their expression during postnatal development, in adulthood, and in the context of cancer. Systemic induction of E2f7 and, to lesser extent, E2f8 transgenes in juvenile mice impaired cell proliferation, caused replication stress, DNA damage, and apoptosis, and inhibited animal growth. In adult mice, however, E2F7 and E2F8 induction was well tolerated, yet profoundly interfered with DNA replication, DNA integrity, and cell proliferation in diethylnitrosamine-induced liver tumors., Conclusion: Collectively, our findings demonstrate that atypical E2Fs can override cell-cycle entry and progression governed by other E2F family members and suggest that this property can be exploited to inhibit proliferation of neoplastic hepatocytes when growth and development have subsided during adulthood., (© 2020 The Authors. Hepatology published by Wiley Periodicals LLC on behalf of American Association for the Study of Liver Diseases.)
- Published
- 2021
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46. PIDDosome-induced p53-dependent ploidy restriction facilitates hepatocarcinogenesis.
- Author
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Sladky VC, Knapp K, Szabo TG, Braun VZ, Bongiovanni L, van den Bos H, Spierings DC, Westendorp B, Curinha A, Stojakovic T, Scharnagl H, Timelthaler G, Tsuchia K, Pinter M, Semmler G, Foijer F, de Bruin A, Reiberger T, Rohr-Udilova N, and Villunger A
- Subjects
- Animals, Carcinogenesis genetics, Humans, Mice, Ploidies, Tumor Suppressor Protein p53 genetics, Carcinoma, Hepatocellular genetics, Liver Neoplasms genetics
- Abstract
Polyploidization frequently precedes tumorigenesis but also occurs during normal development in several tissues. Hepatocyte ploidy is controlled by the PIDDosome during development and regeneration. This multi-protein complex is activated by supernumerary centrosomes to induce p53 and restrict proliferation of polyploid cells, otherwise prone for chromosomal instability. PIDDosome deficiency in the liver results in drastically increased polyploidy. To investigate PIDDosome-induced p53-activation in the pathogenesis of liver cancer, we chemically induced hepatocellular carcinoma (HCC) in mice. Strikingly, PIDDosome deficiency reduced tumor number and burden, despite the inability to activate p53 in polyploid cells. Liver tumors arise primarily from cells with low ploidy, indicating an intrinsic pro-tumorigenic effect of PIDDosome-mediated ploidy restriction. These data suggest that hyperpolyploidization caused by PIDDosome deficiency protects from HCC. Moreover, high tumor cell density, as a surrogate marker of low ploidy, predicts poor survival of HCC patients receiving liver transplantation. Together, we show that the PIDDosome is a potential therapeutic target to manipulate hepatocyte polyploidization for HCC prevention and that tumor cell density may serve as a novel prognostic marker for recurrence-free survival in HCC patients., (© 2020 The Authors. Published under the terms of the CC BY 4.0 license.)
- Published
- 2020
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47. Excessive E2F Transcription in Single Cancer Cells Precludes Transient Cell-Cycle Exit after DNA Damage.
- Author
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Segeren HA, van Rijnberk LM, Moreno E, Riemers FM, van Liere EA, Yuan R, Wubbolts R, de Bruin A, and Westendorp B
- Subjects
- Cell Cycle, Humans, DNA Damage genetics, E2F Transcription Factors metabolism, Sequence Analysis, RNA methods
- Abstract
E2F transcription factors control the expression of cell-cycle genes. Cancers often demonstrate enhanced E2F target gene expression, which can be explained by increased percentages of replicating cells. However, we demonstrate in human cancer biopsy specimens that individual neoplastic cells display abnormally high levels of E2F-dependent transcription. To mimic this situation, we delete the atypical E2F repressors (E2F7/8) or overexpress the E2F3 activator in untransformed cells. Cells with elevated E2F activity during S/G2 phase fail to exit the cell cycle after DNA damage and undergo mitosis. In contrast, wild-type cells complete S phase and then exit the cell cycle by activating the APC/C
Cdh1 via repression of the E2F target Emi1. Many arrested wild-type cells eventually inactivate APC/CCdh1 to execute a second round of DNA replication and mitosis, thereby becoming tetraploid. Cells with elevated E2F transcription fail to exit the cell cycle after DNA damage, which potentially causes genomic instability, promotes malignant progression, and reduces drug sensitivity., Competing Interests: Declaration of Interests The authors declare no competing interests., (Copyright © 2020 The Author(s). Published by Elsevier Inc. All rights reserved.)- Published
- 2020
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48. E2F-Family Members Engage the PIDDosome to Limit Hepatocyte Ploidy in Liver Development and Regeneration.
- Author
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Sladky VC, Knapp K, Soratroi C, Heppke J, Eichin F, Rocamora-Reverte L, Szabo TG, Bongiovanni L, Westendorp B, Moreno E, van Liere EA, Bakker B, Spierings DCJ, Wardenaar R, Pereyra D, Starlinger P, Schultze S, Trauner M, Stojakovic T, Scharnagl H, Fava LL, Foijer F, de Bruin A, and Villunger A
- Subjects
- Aneuploidy, Animals, CRADD Signaling Adaptor Protein physiology, Centrosome, Cyclin-Dependent Kinase Inhibitor p21 physiology, Cytokinesis, Female, Hepatocytes metabolism, Humans, Male, Mice, Mice, Knockout, Caspase 2 physiology, Death Domain Receptor Signaling Adaptor Proteins physiology, E2F Transcription Factors physiology, Hepatocytes cytology, Liver Regeneration, Polyploidy, Tumor Suppressor Protein p53 physiology
- Abstract
E2F transcription factors control the cytokinesis machinery and thereby ploidy in hepatocytes. If or how these proteins limit proliferation of polyploid cells with extra centrosomes remains unknown. Here, we show that the PIDDosome, a signaling platform essential for caspase-2-activation, limits hepatocyte ploidy and is instructed by the E2F network to control p53 in the developing as well as regenerating liver. Casp2 and Pidd1 act as direct transcriptional targets of E2F1 and its antagonists, E2F7 and E2F8, that together co-regulate PIDDosome expression during juvenile liver growth and regeneration. Of note, whereas hepatocyte aneuploidy correlates with the basal ploidy state, the degree of aneuploidy itself is not limited by PIDDosome-dependent p53 activation. Finally, we provide evidence that the same signaling network is engaged to control ploidy in the human liver after resection. Our study defines the PIDDosome as a primary target to manipulate hepatocyte ploidy and proliferation rates in the regenerating liver., Competing Interests: Declaration of Interests The authors declare no competing interests., (Copyright © 2019 Elsevier Inc. All rights reserved.)
- Published
- 2020
- Full Text
- View/download PDF
49. Cyclin F-dependent degradation of E2F7 is critical for DNA repair and G2-phase progression.
- Author
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Yuan R, Liu Q, Segeren HA, Yuniati L, Guardavaccaro D, Lebbink RJ, Westendorp B, and de Bruin A
- Subjects
- Cell Cycle Checkpoints, Cyclins genetics, DNA Damage, DNA Replication, E2F7 Transcription Factor genetics, HeLa Cells, Humans, Protein Binding, Repressor Proteins genetics, Ubiquitin-Protein Ligases genetics, Ubiquitin-Protein Ligases metabolism, Ubiquitination, Cyclins metabolism, DNA Repair, E2F7 Transcription Factor metabolism, G2 Phase physiology, Proteolysis, Repressor Proteins metabolism
- Abstract
E2F7 and E2F8 act as tumor suppressors via transcriptional repression of genes involved in S-phase entry and progression. Previously, we demonstrated that these atypical E2Fs are degraded by APC/C
C dh1 during G1 phase of the cell cycle. However, the mechanism driving the downregulation of atypical E2Fs during G2 phase is unknown. Here, we show that E2F7 is targeted for degradation by the E3 ubiquitin ligase SCFcyclin F during G2. Cyclin F binds via its cyclin domain to a conserved C-terminal CY motif on E2F7. An E2F7 mutant unable to interact with SCFcyclin F remains stable during G2. Furthermore, SCFcyclin F can also interact and induce degradation of E2F8. However, this does not require the cyclin domain of SCFcyclin F nor the CY motifs in the C-terminus of E2F8, implying a different regulatory mechanism than for E2F7. Importantly, depletion of cyclin F causes an atypical-E2F-dependent delay of the G2/M transition, accompanied by reduced expression of E2F target genes involved in DNA repair. Live cell imaging of DNA damage revealed that cyclin F-dependent regulation of atypical E2Fs is critical for efficient DNA repair and cell cycle progression., (© 2019 The Authors. Published under the terms of the CC BY 4.0 license.)- Published
- 2019
- Full Text
- View/download PDF
50. DYRK1A Is a Regulator of S-Phase Entry in Hepatic Progenitor Cells.
- Author
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Kruitwagen HS, Westendorp B, Viebahn CS, Post K, van Wolferen ME, Oosterhoff LA, Egan DA, Delabar JM, Toussaint MJ, Schotanus BA, de Bruin A, Rothuizen J, Penning LC, and Spee B
- Subjects
- Adult Stem Cells cytology, Cell Line, Humans, Liver cytology, Protein Serine-Threonine Kinases genetics, Protein-Tyrosine Kinases genetics, Dyrk Kinases, Adult Stem Cells metabolism, Liver metabolism, Protein Serine-Threonine Kinases biosynthesis, Protein-Tyrosine Kinases biosynthesis, S Phase physiology, Transcription, Genetic physiology
- Abstract
Hepatic progenitor cells (HPCs) are adult liver stem cells that act as second line of defense in liver regeneration. They are normally quiescent, but in case of severe liver damage, HPC proliferation is triggered by external activation mechanisms from their niche. Although several important proproliferative mechanisms have been described, it is not known which key intracellular regulators govern the switch between HPC quiescence and active cell cycle. We performed a high-throughput kinome small interfering RNA (siRNA) screen in HepaRG cells, a HPC-like cell line, and evaluated the effect on proliferation with a 5-ethynyl-2'-deoxyuridine (EdU) incorporation assay. One hit increased the percentage of EdU-positive cells after knockdown: dual specificity tyrosine phosphorylation regulated kinase 1A (DYRK1A). Although upon DYRK1A silencing, the percentage of EdU- and phosphorylated histone H3 (pH3)-positive cells was increased, and total cell numbers were not increased, possibly through a subsequent delay in cell cycle progression. This phenotype was confirmed with chemical inhibition of DYRK1A using harmine and with primary HPCs cultured as liver organoids. DYRK1A inhibition impaired Dimerization Partner, RB-like, E2F, and multivulva class B (DREAM) complex formation in HPCs and abolished its transcriptional repression on cell cycle progression. To further analyze DYRK1A function in HPC proliferation, liver organoid cultures were established from mBACtgDyrk1A mice, which harbor one extra copy of the murine Dyrk1a gene (Dyrk+++). Dyrk+++ organoids had both a reduced percentage of EdU-positive cells and reduced proliferation compared with wild-type organoids. This study provides evidence for an essential role of DYRK1A as balanced regulator of S-phase entry in HPCs. An exact gene dosage is crucial, as both DYRK1A deficiency and overexpression affect HPC cell cycle progression.
- Published
- 2018
- Full Text
- View/download PDF
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