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Epigenetic Regulators Rbbp4 and Hdac1 Are Overexpressed Genetics, Development and Cell Biology Genetics, Development and Cell Biology Publications 2018 Epigenetic regulators Rbbp4 and Hdac1 are overexpressed in a zebrafish model of RB1 embryonal brain tumor, and are required for neural progenitor survival and proliferation Laura E. Schultz Iowa State University Jeffrey A. Haltom Iowa State University, [email protected] Maira P. Almeida Iowa State University, [email protected] Wesley A. Wierson Iowa State University SFtoalcloi Lw. thiSolins and additional works at: https://lib.dr.iastate.edu/gdcb_las_pubs IowaP Satrate of U ntheiversitCyancer Biology Commons, Genetics Commons, and the Genomics Commons TheSee nex tompc page forle addte bitioniblaiol agruthorapshic information for this item can be found at https://lib.dr.iastate.edu/ gdcb_las_pubs/206. For information on how to cite this item, please visit http://lib.dr.iastate.edu/ howtocite.html. This Article is brought to you for free and open access by the Genetics, Development and Cell Biology at Iowa State University Digital Repository. It has been accepted for inclusion in Genetics, Development and Cell Biology Publications by an authorized administrator of Iowa State University Digital Repository. For more information, please contact [email protected]. Epigenetic regulators Rbbp4 and Hdac1 are overexpressed in a zebrafish model of RB1 embryonal brain tumor, and are required for neural progenitor survival and proliferation Abstract In this study, we used comparative genomics and developmental genetics to identify epigenetic regulators driving oncogenesis in a zebrafish retinoblastoma 1 (rb1) somatic-targeting model of RB1 mutant embryonal brain tumors. Zebrafish rb1 brain tumors caused by TALEN or CRISPR targeting are histologically similar to human central nervous system primitive neuroectodermal tumors (CNS-PNETs). Like the human oligoneural OLIG2+/SOX10+ CNS-PNET subtype, zebrafish rb1 tumors show elevated expression of neural progenitor transcription factors olig2, sox10, sox8b and the receptor tyrosine kinase erbb3a oncogene. Comparison of rb1 tumor and rb1/rb1 germline mutant larval transcriptomes shows that the altered oligoneural precursor signature is specific ot tumor tissue. More than 170 chromatin regulators were differentially expressed in rb1 tumors, including overexpression of chromatin remodeler components histone deacetylase 1 (hdac1) and retinoblastoma binding protein 4(rbbp4). Germline mutant analysis confirms that zebrafish rb1, rbbp4 and hdac1 are required during brain development. rb1 is necessary for neural precursor cell cycle exit and terminal differentiation, rbbp4 is required for survival of postmitotic precursors, and hdac1 maintains proliferation of the neural stem cell/progenitor pool. We present an in vivo assay using somatic CRISPR targeting plus live imaging of histone-H2A.F/Z-GFP fusion protein in developing larval brain to rapidly test the role of chromatin remodelers in neural stem and progenitor cells. Our somatic assay recapitulates germline mutant phenotypes and reveals a dynamic view of their roles in neural cell populations. Our study provides new insight into the epigenetic processes that might drive pathogenesis in RB1 brain tumors, and identifies Rbbp4 and its associated chromatin remodeling complexes as potential target pathways to induce apoptosis in RB1 mutant brain cancer cells. Keywords Zebrafish, Rbbp4, Hdac1, RB1 embryonal oligoneural brain tumor, Epigenetic regulators, CRISPR somatic targeting, Neural progenitor Disciplines Cancer Biology | Cell and Developmental Biology | Genetics | Genomics Comments This article is published as Schultz, Laura E., Jeffrey A. Haltom, Maira P. Almeida, Wesley A. Wierson, Staci L. Solin, Trevor J. Weiss, Jordan A. Helmer, Elizabeth J. Sandquist, Heather R. Shive, and Maura McGrail. "Epigenetic regulators Rbbp4 and Hdac1 are overexpressed in a zebrafish model of RB1 embryonal brain tumor, and are required for neural progenitor survival and proliferation." Disease models & mechanisms 11, no. 6 (2018): dmm034124. doi: 10.1242/dmm.034124. Creative Commons License This work is licensed under a Creative Commons Attribution 3.0 License. This article is available at Iowa State University Digital Repository: https://lib.dr.iastate.edu/gdcb_las_pubs/206 Authors Laura E. Schultz, Jeffrey A. Haltom, Maira P. Almeida, Wesley A. Wierson, Staci L. Solin, Trevor J. Weiss, Jordan A. Helmer, Elizabeth J. Sandquist, Heather R. Shive, and Maura McGrail This article is available at Iowa State University Digital Repository: https://lib.dr.iastate.edu/gdcb_las_pubs/206 © 2018. Published by The Company of Biologists Ltd | Disease Models & Mechanisms (2018) 11, dmm034124. doi:10.1242/dmm.034124 RESEARCH ARTICLE Epigenetic regulators Rbbp4 and Hdac1 are overexpressed in a zebrafish model of RB1 embryonal brain tumor, and are required for neural progenitor survival and proliferation Laura E. Schultz1, Jeffrey A. Haltom1, Maira P. Almeida1, Wesley A. Wierson1, Staci L. Solin1,*, Trevor J. Weiss1, Jordan A. Helmer1, Elizabeth J. Sandquist1, Heather R. Shive2 and Maura McGrail1,‡ ABSTRACT INTRODUCTION In this study, we used comparative genomics and developmental The retinoblastoma tumor suppressor Retinoblastoma 1 (RB1) plays genetics to identify epigenetic regulators driving oncogenesis in a distinct roles in regulating proliferation and differentiation in stem, zebrafish retinoblastoma 1 (rb1) somatic-targeting model of RB1 progenitor and lineage-restricted cell populations (Fong and Slack, mutant embryonal brain tumors. Zebrafish rb1 brain tumors caused by 2017; Julian and Blais, 2015; Sage, 2012). The canonical tumor TALEN or CRISPR targeting are histologically similar to human central suppressor role of RB1 is to regulate proliferation by transcriptional nervous system primitive neuroectodermal tumors (CNS-PNETs). repression of E2F targets driving cell cycle entry (Dyson, 2016). RB1 Like the human oligoneural OLIG2+/SOX10+ CNS-PNET subtype, might also function as a tumor suppressor by promoting differentiation zebrafish rb1 tumors show elevated expression of neural progenitor of lineage-committed cells (Calo et al., 2010) and survival of transcription factors olig2, sox10, sox8b and the receptor tyrosine postmitotic neurons (Naser et al., 2016; Vandenbosch et al., 2016), kinase erbb3a oncogene. Comparison of rb1 tumor and rb1/rb1 and preventing cellular reprogramming (Kareta et al., 2015). RB1 germline mutant larval transcriptomes shows that the altered transcriptional control is mediated, in part, through association with oligoneural precursor signature is specific to tumor tissue. More than chromatin remodelers that modify post-translational marks on histones 170 chromatin regulators were differentially expressed in rb1 tumors, to activate or repress gene expression (Beshiri et al., 2012; Brehm et al., including overexpression of chromatin remodeler components histone 1998; Lin et al., 2011). Like RB1, many chromatin remodelers are deacetylase 1 (hdac1)andretinoblastoma binding protein 4 (rbbp4). mutated in human cancers (Vogelstein et al., 2013), implicating Germline mutant analysis confirms that zebrafish rb1, rbbp4 and hdac1 epigenetic control of gene expression as a significant contributor to are required during brain development. rb1 is necessary for neural oncogenesis (Conway et al., 2015; Suva et al., 2013). Recent analyses precursor cell cycle exit and terminal differentiation, rbbp4 is required of the epigenome in human and mouse retinoblastoma show that for survival of postmitotic precursors, and hdac1 maintains proliferation histone modification correlates with oncogenic gene expression (Aldiri of the neural stem cell/progenitor pool. We present an in vivo assay et al., 2017; Benavente et al., 2013; Zhang et al., 2012). Understanding using somatic CRISPR targeting plus live imaging of histone-H2A.F/ how RB1 loss affects epigenetic control in neural stem and progenitor Z-GFP fusion protein in developing larval brain to rapidly test the role of cells during development and in brain cancer is important for chromatin remodelers in neural stem and progenitor cells. Our somatic identifying new pathways that contribute to carcinogenesis. assay recapitulates germline mutant phenotypes and reveals a Two epigenetic regulators that directly interact with RB1 are the dynamic view of their roles in neural cell populations. Our study retinoblastoma binding protein RBBP4 (Qian and Lee, 1995; Qian provides new insight into the epigenetic processes that might drive et al., 1993) and the Class I histone deacetylase HDAC1 (Brehm et al., pathogenesis in RB1 brain tumors, and identifies Rbbp4 and its 1998; Luo et al., 1998; Magnaghi-Jaulin et al., 1998). RBBP4 is a associated chromatin remodeling complexes as potential target WD40-repeat histone chaperone (Burkhart and Sage, 2008; Murzina pathways to induce apoptosis in RB1 mutant brain cancer cells. et al., 2008; Nowak et al., 2011; Song et al., 2008) and a chromatin adaptor component of multiple remodeling complexes, including the This article has an associated First Person interview with the first nucleosome remodeling and histone deacetylase complex (NuRD) author of the paper. (Alqarni et al., 2014) and the cell cycle regulatory DREAM complex (Harrison et al., 2006; Litovchick et al., 2007). RBBP4 expression is KEY WORDS: Zebrafish, Rbbp4, Hdac1, RB1 embryonal oligoneural altered in human glioma (Forbes et al., 2017), and was recently shown brain tumor, Epigenetic regulators, CRISPR somatic targeting, to cooperate
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