DOCSLIB.ORG

Enhancers and Super-Enhancers Have an Equivalent Regulatory Role in Embryonic Stem Cells Through Regulation of Single Or Multiple Genes

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Enhancers and Super-Enhancers Have an Equivalent Regulatory Role in Embryonic Stem Cells Through Regulation of Single Or Multiple Genes Downloaded from genome.cshlp.org on October 6, 2021 - Published by Cold Spring Harbor Laboratory Press Enhancers and super-enhancers have an equivalent regulatory role in embryonic stem cells through regulation of single or multiple genes Authors: Sakthi D Moorthy*1, Scott Davidson*1, Virlana M Shchuka1, Gurdeep Singh1, Nakisa Malek- Gilani1, Lida Langroudi1, Alexandre Martchenko1, Vincent So1, Neil N Macpherson1, and Jennifer A Mitchell1,2. * These authors contributed equally 1) Department of Cell and Systems Biology, University of Toronto, Toronto, ON, M5S 3G5, Canada. 2) Centre for the Analysis of Genome Evolution and Function, University of Toronto, Toronto, ON, M5S 3G5, Canada. Contact: Jennifer A Mitchell, Department of Cell and Systems Biology, University of Toronto, 25 Harbord Street, Toronto, Ontario M5S 3G5, Canada. Phone: (416)978-6711, Fax: (416)978-8532, Email: [email protected]. Running Title: Enhancer and super-enhancer equivalence Keywords: Transcription, Enhancer Elements, Super-enhancers, Embryonic Stem Cells, Gene Expression Regulation. 1 Downloaded from genome.cshlp.org on October 6, 2021 - Published by Cold Spring Harbor Laboratory Press ABSTRACT Transcriptional enhancers are critical for maintaining cell type-specific gene expression and driving cell fate changes during development. Highly transcribed genes are often associated with a cluster of individual enhancers such as those found in locus control regions. Recently these have been termed stretch enhancers or super-enhancers, which have been predicted to regulate critical cell identity genes. We employed a CRISPR/Cas9-mediated deletion approach to study the function of several enhancer clusters (ECs) and isolated enhancers in mouse embryonic stem (ES) cells. Our results reveal that the effect of deleting ECs, also classified as ES cell super-enhancers, is highly variable, resulting in target gene expression reductions ranging from 12% to as much as 92%. Partial deletions of these ECs which removed only one enhancer or a sub-cluster of enhancers revealed partially redundant control of the regulated gene by multiple enhancers within the larger cluster. Many highly transcribed genes in ES cells are not associated with a super-enhancer; furthermore, super-enhancer predictions ignore 81% of the potentially active regulatory elements predicted by co-binding of 5 or more pluripotency- associated transcription factors. Deletion of these additional enhancer regions revealed their robust regulatory role in gene transcription. In addition, select super-enhancers and enhancers were identified which regulated clusters of paralogous genes. We conclude that whereas robust transcriptional output can be achieved by an isolated enhancer, clusters of enhancers acting on a common target gene act in a partially redundant manner to fine tune transcriptional output of their target genes. INTRODUCTION Distal regulatory elements are critical in establishing and maintaining tissue-specific transcriptional regulation of gene expression and are central in controlling cell identity. Furthermore, genome-wide association studies identifying disease and phenotypic trait- associated variants have found that the majority of these variants are within non-coding regions of the genome, suggesting regulatory activity (Maurano et al., 2012; Schaub et al., 2012). Further supporting this hypothesis are the observations that these non-coding regions overlap DNase I hypersensitive sites (DHSs) found in accessible chromatin and that the disease- 2 Downloaded from genome.cshlp.org on October 6, 2021 - Published by Cold Spring Harbor Laboratory Press associated variants often disrupt existing or create new transcription factor binding motifs. Alterations in regulatory DNA by large deletion or single mutation are known to cause disease and phenotypic alterations. For example, deletion of the hemoglobin subunit beta (HBB) locus control region (LCR) 50 kb upstream of the HBB gene causes β-thalassemia due to the inability of erythroid cells to produce mature globin (Kioussis et al., 1983; Tuan et al., 1989). As these critical regulatory sequences can be located tens to hundreds of kb away from the gene or genes they regulate, it can be challenging to identify regulatory elements and their target genes. A striking example of this is the limb enhancer that regulates the SHH (sonic hedgehog) gene. This enhancer is located 1 Mb away from the SHH gene in the intron of the LMBR1 gene and point mutations in this enhancer are associated with preaxial polydactyly in humans (Lettice et al., 2003). These findings highlight the necessity for a mechanistic understanding of the distal regulatory elements that regulate gene transcription. Several approaches have been used to identify transcriptional enhancers including: sequence based approaches that rely on transcription factor motif identification, sequence conservation through evolution, co-activator binding (EP300 and Mediator), histone modifications, open chromatin and approaches that combine multiple such features (Ballester et al., 2014; Chen et al., 2012; Ernst and Kellis, 2012; Hallikas et al., 2006; Heintzman et al., 2007; Hoffman et al., 2012; Pennacchio et al., 2006; Visel et al., 2009). Despite these numerous efforts and the wealth of data generated by the ENCODE Project (Bernstein et al., 2012), only a fraction (26%) of enhancer predictions display enhancer activity in reporter assays (Kwasnieski et al., 2014). Mid- to high-throughput screening methods such as MPRA, STARR-seq, and FIREWACh, facilitate testing the activity of enhancers genome-wide in transiently, or in the case of FIREWACh stably, transfected cell lines (Arnold et al., 2013; Melnikov et al., 2012; Murtha et al., 2014); however, these assays do not test activity in the endogenous genomic context and therefore do not identify the regulated gene. Recently the concept of super-enhancers and stretch enhancers was proposed and these regions were predicted to regulate cell identity genes and to confer higher expression on their target genes compared to regions termed typical enhancers (Hnisz et al., 2013; Loven et al., 2013; Parker et al., 2013; Whyte et al., 2013). Stretch enhancers are regions of ≥3 kb which 3 Downloaded from genome.cshlp.org on October 6, 2021 - Published by Cold Spring Harbor Laboratory Press exhibit an enhancer chromatin state based on the ChromHMM algorithm (Ernst et al., 2011; Parker et al., 2013). Super-enhancers have been predicted in numerous cell types where these regions have been identified by increased binding of the mediator protein MED1, enrichment of histone H3 K27 acetylation (H3K27ac), or binding of cell type-specific transcription factors (Khan and Zhang, 2016). The super-enhancer prediction involves bioinformatically stitching regions within 12.5 kb of one another in the linear genome; as a result super-enhancers are on average longer than traditional enhancers with a median size of 8.7 kb, and often contain more than one separate region which is bound by multiple transcription factors (Hnisz et al., 2013; Loven et al., 2013; Whyte et al., 2013). Although super-enhancers have been predicted in numerous cell types, few have been functionally investigated by deletion and it remains unclear whether these predicted super-enhancers or stretch enhancers represent a novel paradigm in gene regulation separate from the traditional concept of enhancers (Pott and Lieb, 2014). In fact the two classic robust distal regulatory elements, the LCRs of the Hbb and hemoglobin subunit alpha (Hba) genes, are strikingly similar to super-enhancers; they contain multiple DHSs or are enhancer clusters (ECs), with each enhancer bound by multiple transcription factors (Chen et al., 1997; Grosveld et al., 1987; Higgs et al., 1990; Mitchell et al., 2012; Tuan et al., 1989). Furthermore, both the Hbb and Hba LCRs were recently determined to fit the bioinformatic criteria for super-enhancers (Hay et al., 2016). The transcriptional regulatory networks that control cell identity of embryonic stem (ES) cells have been intensely studied making mouse ES cells an ideal model for the study of transcriptional regulation. This knowledge has led to the development of numerous reprogramming strategies to generate induced pluripotent stem (iPS) cells in multiple species. Despite this knowledge, we lack a clear understanding of where in the genome the critical regulatory elements regulating pluripotency lie and how those elements function in their endogenous context. Advances in genome editing technology have provided the opportunity to characterize this regulatory landscape with greater fidelity. One example of this approach applied to ES cells is deletion of the enhancers surrounding the Sox2 gene. Two studies revealed that a cluster of predicted enhancer regions 100 kb downstream of the Sox2 gene, termed the Sox2 control region (SCR), are required for Sox2 transcription in mouse ES cells (Li et al., 2014; 4 Downloaded from genome.cshlp.org on October 6, 2021 - Published by Cold Spring Harbor Laboratory Press Zhou et al., 2014). The SCR overlaps a predicted super-enhancer and was found to be required for >85% of the transcript levels of Sox2 in ES cells. Similarly, other studies have revealed the role of select super-enhancers in gene regulation (Hay et al., 2016; Hnisz et al., 2015; Huang et al., 2016; Mansour et al., 2014; Shin et al., 2016). At the Sox2 locus an additional super- enhancer was predicted surrounding the Sox2 gene; however, deletion of the enhancers within that region
Load more

Recommended publications
  • Hedgehog Signaling Is Evolutionarily Conserved Cilium-Independent
    Downloaded from genesdev.cshlp.org on August 14, 2009 - Published by Cold Spring Harbor Laboratory Press Cilium-independent regulation of Gli protein function by Sufu in Hedgehog signaling is evolutionarily conserved Miao-Hsueh Chen, Christopher W. Wilson, Ya-Jun Li, et al. Genes Dev. 2009 23: 1910-1928 Access the most recent version at doi:10.1101/gad.1794109 Supplemental http://genesdev.cshlp.org/content/suppl/2009/07/23/23.16.1910.DC1.html Material References This article cites 97 articles, 47 of which can be accessed free at: http://genesdev.cshlp.org/content/23/16/1910.full.html#ref-list-1 Email alerting Receive free email alerts when new articles cite this article - sign up in the box at the service top right corner of the article or click here To subscribe to Genes & Development go to: http://genesdev.cshlp.org/subscriptions Copyright © 2009 by Cold Spring Harbor Laboratory Press Downloaded from genesdev.cshlp.org on August 14, 2009 - Published by Cold Spring Harbor Laboratory Press Cilium-independent regulation of Gli protein function by Sufu in Hedgehog signaling is evolutionarily conserved Miao-Hsueh Chen,1,3 Christopher W. Wilson,1,3 Ya-Jun Li,1 Kelvin King Lo Law,2 Chi-Sheng Lu,1 Rhodora Gacayan,1 Xiaoyun Zhang,2 Chi-chung Hui,2 and Pao-Tien Chuang1,4 1Cardiovascular Research Institute, University of California at San Francisco, San Francisco, California 94158, USA; 2Program in Developmental and Stem Cell Biology, The Hospital for Sick Children, and Department of Molecular Genetics, University of Toronto, Toronto, Ontario M5G 1L7, Canada A central question in Hedgehog (Hh) signaling is how evolutionarily conserved components of the pathway might use the primary cilium in mammals but not fly.
    [Show full text]
  • Gli Transcription Factors Mediate the Oncogenic Transformation of Prostate Basal Cells Induced by a Kras-Androgen Receptor Axis*
    crossmark THE JOURNAL OF BIOLOGICAL CHEMISTRY VOL. 291, NO. 49, pp. 25749–25760, December 2, 2016 © 2016 by The American Society for Biochemistry and Molecular Biology, Inc. Published in the U.S.A. Gli Transcription Factors Mediate the Oncogenic Transformation of Prostate Basal Cells Induced by a Kras-Androgen Receptor Axis*□S Received for publication, August 12, 2016, and in revised form, September 28, 2016 Published, JBC Papers in Press, October 19, 2016, DOI 10.1074/jbc.M116.753129 Meng Wu‡, Lishann Ingram‡, Ezequiel J. Tolosa§, Renzo E. Vera§, Qianjin Li‡, Sungjin Kim‡, Yongjie Ma‡, Demetri D. Spyropoulos¶, Zanna Beharryʈ, Jiaoti Huang**, Martin E. Fernandez-Zapico§, and Houjian Cai‡1 From the ‡Department of Pharmaceutical and Biomedical Sciences, College of Pharmacy, University of Georgia, Athens, Georgia 30602, the §Schulze Center for Novel Therapeutics, Division of Oncology Research, Mayo Clinic, Rochester, Minnesota 55905, the ¶Department of Pathology and Laboratory Medicine, Medical University of South Carolina, Charleston, South Carolina 29425, the ʈDepartment of Chemistry and Physics, Florida Gulf Coast University, Fort Myers, Florida 33965, and the **Department of Pathology, School of Medicine, Duke University, Durham, North Carolina 27710 Edited by Eric Fearon Although the differentiation of oncogenically transformed cer progression has been characterized with multiple stages, 2 basal progenitor cells is one of the key steps in prostate tumori- including benign, prostatic intraepithelial neoplasia (PIN), Downloaded from genesis, the mechanisms mediating this cellular process are still invasive adenocarcinoma, and metastatic cancer (2). Numerous ؉ largely unknown. Here we demonstrate that an expanded p63 oncogenic driver genes, including loss of tumor suppressors, ؉ and CK5 basal/progenitor cell population, induced by the con- overexpression, and/or activation of oncogenes, have been comitant activation of oncogenic Kras(G12D) and androgen identified based on genetic analysis of clinical prostate tumors.
    [Show full text]
  • Cross-Talk Between HER2 and MED1 Regulates Tamoxifen Resistance of Human Breast Cancer Cells
    Published OnlineFirst September 10, 2012; DOI: 10.1158/0008-5472.CAN-12-1305 Cancer Tumor and Stem Cell Biology Research Cross-talk between HER2 and MED1 Regulates Tamoxifen Resistance of Human Breast Cancer Cells Jiajun Cui1, Katherine Germer1, Tianying Wu2, Jiang Wang3, Jia Luo5, Shao-chun Wang1, Qianben Wang4, and Xiaoting Zhang1 Abstract Despite the fact that most breast cancer patients have estrogen receptor (ER) a-positive tumors, up to 50% of the patients are or soon develop resistance to endocrine therapy. It is recognized that HER2 activation is one of the major mechanisms contributing to endocrine resistance. In this study, we report that the ER coactivator MED1 is a novel cross-talk point for the HER2 and ERa pathways. Tissue microarray analysis of human breast cancers revealed that MED1 expression positively correlates most strongly with HER2 status of the tumors. MED1 was highly phosphorylated, in a HER2-dependent manner, at the site known to be critical for its activation. Importantly, RNAi-mediated attenuation of MED1 sensitized HER2-overexpressing cells to tamoxifen treatment. MED1 and its phosphorylated form, but not the corepressors N-CoR and SMRT, were recruited to the ERa target gene promoter by tamoxifen in HER2-overexpressing cells. Significantly, MED1 attenuation or mutation of MED1 phosphorylation sites was sufficient to restore the promoter recruitment of N-CoR and SMRT. Notably, we found that MED1 is required for the expression of not only traditional E2-ERa target genes but also the newly described EGF-ERa target genes. Our results additionally indicated that MED1 is recruited to the HER2 gene and required for its expression.
    [Show full text]
  • Key Roles for MED1 Lxxll Motifs in Pubertal Mammary Gland Development and Luminal-Cell Differentiation
    Key roles for MED1 LxxLL motifs in pubertal mammary gland development and luminal-cell differentiation Pingping Jianga,b,1, Qiuping Hua,1, Mitsuhiro Itoc,1,3, Sara Meyera, Susan Waltza, Sohaib Khana, Robert G. Roederc,2, and Xiaoting Zhanga,2 aDepartment of Cancer and Cell Biology, College of Medicine, University of Cincinnati, 3125 Eden Avenue, OH 45267; bCollege of Life Sciences, Zhejiang University, 388 Yuhangtang Road, Hangzhou 310058, China; and cLaboratory of Biochemistry and Molecular Biology, The Rockefeller University, 1230 York Avenue, New York, NY 10065 Contributed by Robert G. Roeder, February 16, 2010 (sent for review January 18, 2010) Mediator recently has emerged as a central player in the direct (7–9). In general, individual Mediator subunits interact transduction of signals from transcription factors to the general specifically with their corresponding transcription factors and de- transcriptional machinery. In the case of nuclear receptors, in vitro letions of these Mediator subunits often affect expression primar- studies have shown that the transcriptional coactivator function of ily of target genes and pathways controlled by their corresponding the Mediator involves direct ligand-dependent interactions of the transcription factor(s) (9). In the case of nuclear receptors, the MED1 subunit, through its two classical LxxLL motifs, with the re- Mediator interactions are ligand- and AF-2-dependent and ceptor AF2 domain. However, despite the strong in vitro evidence, mediated through the LxxLL motifs in the MED1 (a.k.a. there currently is little information regarding in vivo functions of TRAP220/PBP/DRIP205) subunit (10–13). Importantly, in rela- the LxxLL motifs either in MED1 or in other coactivators.
    [Show full text]
  • Non-Canonical Activation of Hedgehog in Prostate Cancer Cells Mediated by the Interaction of Transcriptionally Active Androgen Receptor Proteins with Gli3
    Oncogene (2018) 37:2313–2325 https://doi.org/10.1038/s41388-017-0098-7 ARTICLE Non-canonical activation of hedgehog in prostate cancer cells mediated by the interaction of transcriptionally active androgen receptor proteins with Gli3 1 1,2 2 1 1 1 2,3 Na Li ● Sarah Truong ● Mannan Nouri ● Jackson Moore ● Nader Al Nakouzi ● Amy Anne Lubik ● Ralph Buttyan Received: 19 July 2017 / Revised: 18 October 2017 / Accepted: 29 November 2017 / Published online: 12 February 2018 © The Author(s) 2018. This article is published with open access Abstract Hedgehog (Hh) is an oncogenic signaling pathway that regulates the activity of Gli transcription factors. Canonical Hh is a Smoothened-(Smo-) driven process that alters the post-translational processing of Gli2/Gli3 proteins. Though evidence supports a role for Gli action in prostate cancer (PCa) cell growth and progression, there is little indication that Smo is involved. Here we describe a non-canonical means for activation of Gli transcription in PCa cells mediated by the binding of transcriptionally-active androgen receptors (ARs) to Gli3. Androgens stimulated reporter expression from a Gli-dependent promoter in a variety of AR + PCa cells and this activity was suppressed by an anti-androgen, Enz, or by AR knockdown. 1234567890();,: Androgens also upregulated expression of endogenous Gli-dependent genes. This activity was associated with increased intranuclear binding of Gli3 to AR that was antagonized by Enz. Fine mapping of the AR binding domain on Gli2 showed that AR recognizes the Gli protein processing domain (PPD) in the C-terminus. Mutations in the arginine-/serine repeat elements of the Gli2 PPD involved in phosphorylation and ubiquitinylation blocked the binding to AR.
    [Show full text]
  • Tamoxifen Resistance: Emerging Molecular Targets
    International Journal of Molecular Sciences Review Tamoxifen Resistance: Emerging Molecular Targets Milena Rondón-Lagos 1,*,†, Victoria E. Villegas 2,3,*,†, Nelson Rangel 1,2,3, Magda Carolina Sánchez 2 and Peter G. Zaphiropoulos 4 1 Department of Medical Sciences, University of Turin, Turin 10126, Italy; [email protected] 2 Faculty of Natural Sciences and Mathematics, Universidad del Rosario, Bogotá 11001000, Colombia; [email protected] 3 Doctoral Program in Biomedical Sciences, Universidad del Rosario, Bogotá 11001000, Colombia 4 Department of Biosciences and Nutrition, Karolinska Institutet, Huddinge 14183, Sweden; [email protected] * Correspondence: [email protected] (M.R.-L.); [email protected] (V.E.V.); Tel.: +39-01-1633-4127 (ext. 4388) (M.R.-L.); +57-1-297-0200 (ext. 4029) (V.E.V.); Fax: +39-01-1663-5267 (M.R.-L.); +57-1-297-0200 (V.E.V.) † These authors contributed equally to this work. Academic Editor: William Chi-shing Cho Received: 5 July 2016; Accepted: 16 August 2016; Published: 19 August 2016 Abstract: 17β-Estradiol (E2) plays a pivotal role in the development and progression of breast cancer. As a result, blockade of the E2 signal through either tamoxifen (TAM) or aromatase inhibitors is an important therapeutic strategy to treat or prevent estrogen receptor (ER) positive breast cancer. However, resistance to TAM is the major obstacle in endocrine therapy. This resistance occurs either de novo or is acquired after an initial beneficial response. The underlying mechanisms for TAM resistance are probably multifactorial and remain largely unknown. Considering that breast cancer is a very heterogeneous disease and patients respond differently to treatment, the molecular analysis of TAM’s biological activity could provide the necessary framework to understand the complex effects of this drug in target cells.
    [Show full text]
  • The Structure-Function Relationship of Angular Estrogens and Estrogen Receptor Alpha to Initiate Estrogen-Induced Apoptosis in Breast Cancer Cells S
    Supplemental material to this article can be found at: http://molpharm.aspetjournals.org/content/suppl/2020/05/03/mol.120.119776.DC1 1521-0111/98/1/24–37$35.00 https://doi.org/10.1124/mol.120.119776 MOLECULAR PHARMACOLOGY Mol Pharmacol 98:24–37, July 2020 Copyright ª 2020 The Author(s) This is an open access article distributed under the CC BY Attribution 4.0 International license. The Structure-Function Relationship of Angular Estrogens and Estrogen Receptor Alpha to Initiate Estrogen-Induced Apoptosis in Breast Cancer Cells s Philipp Y. Maximov, Balkees Abderrahman, Yousef M. Hawsawi, Yue Chen, Charles E. Foulds, Antrix Jain, Anna Malovannaya, Ping Fan, Ramona F. Curpan, Ross Han, Sean W. Fanning, Bradley M. Broom, Daniela M. Quintana Rincon, Jeffery A. Greenland, Geoffrey L. Greene, and V. Craig Jordan Downloaded from Departments of Breast Medical Oncology (P.Y.M., B.A., P.F., D.M.Q.R., J.A.G., V.C.J.) and Computational Biology and Bioinformatics (B.M.B.), University of Texas, MD Anderson Cancer Center, Houston, Texas; King Faisal Specialist Hospital and Research (Gen.Org.), Research Center, Jeddah, Kingdom of Saudi Arabia (Y.M.H.); The Ben May Department for Cancer Research, University of Chicago, Chicago, Illinois (R.H., S.W.F., G.L.G.); Center for Precision Environmental Health and Department of Molecular and Cellular Biology (C.E.F.), Mass Spectrometry Proteomics Core (A.J., A.M.), Verna and Marrs McLean Department of Biochemistry and Molecular Biology, Mass Spectrometry Proteomics Core (A.M.), and Dan L. Duncan molpharm.aspetjournals.org
    [Show full text]
  • GATA3 Acts Upstream of FOXA1 in Mediating ESR1 Binding by Shaping Enhancer Accessibility
    Downloaded from genome.cshlp.org on October 3, 2021 - Published by Cold Spring Harbor Laboratory Press Research GATA3 acts upstream of FOXA1 in mediating ESR1 binding by shaping enhancer accessibility Vasiliki Theodorou,1 Rory Stark,2 Suraj Menon,2 and Jason S. Carroll1,3,4 1Nuclear Receptor Transcription Lab, 2Bioinformatics Core, Cancer Research UK, Cambridge Research Institute, Li Ka Shing Centre, Cambridge CB2 0RE, United Kingdom; 3Department of Oncology, University of Cambridge, Cambridge CB2 OXZ, United Kingdom Estrogen receptor (ESR1) drives growth in the majority of human breast cancers by binding to regulatory elements and inducing transcription events that promote tumor growth. Differences in enhancer occupancy by ESR1 contribute to the diverse expression profiles and clinical outcome observed in breast cancer patients. GATA3 is an ESR1-cooperating transcription factor mutated in breast tumors; however, its genomic properties are not fully defined. In order to investigate the composition of enhancers involved in estrogen-induced transcription and the potential role of GATA3, we performed extensive ChIP-sequencing in unstimulated breast cancer cells and following estrogen treatment. We find that GATA3 is pivotal in mediating enhancer accessibility at regulatory regions involved in ESR1-mediated transcription. GATA3 silencing resulted in a global redistribution of cofactors and active histone marks prior to estrogen stimulation. These global genomic changes altered the ESR1-binding profile that subsequently occurred following estrogen, with events exhibiting both loss and gain in binding affinity, implying a GATA3-mediated redistribution of ESR1 binding. The GATA3-mediated redistributed ESR1 profile correlated with changes in gene expression, suggestive of its functionality. Chromatin loops at the TFF locus involving ESR1-bound enhancers occurred independently of ESR1 when GATA3 was silenced, indicating that GATA3, when present on the chromatin, may serve as a licensing factor for estrogen–ESR1-mediated interactions between cis-regulatory elements.
    [Show full text]
  • GLI2 but Not GLI1/GLI3 Plays a Central Role in the Induction of Malignant Phenotype of Gallbladder Cancer
    ONCOLOGY REPORTS 45: 997-1010, 2021 GLI2 but not GLI1/GLI3 plays a central role in the induction of malignant phenotype of gallbladder cancer SHU ICHIMIYA1, HIDEYA ONISHI1, SHINJIRO NAGAO1, SATOKO KOGA1, KUKIKO SAKIHAMA2, KAZUNORI NAKAYAMA1, AKIKO FUJIMURA3, YASUHIRO OYAMA4, AKIRA IMAIZUMI1, YOSHINAO ODA2 and MASAFUMI NAKAMURA4 Departments of 1Cancer Therapy and Research, 2Anatomical Pathology, 3Otorhinolaryngology and 4Surgery and Oncology, Graduate School of Medical Sciences, Kyushu University, Fukuoka 812‑8582, Japan Received August 10, 2020; Accepted December 7, 2020 DOI: 10.3892/or.2021.7947 Abstract. We previously reported that Hedgehog (Hh) signal Introduction was enhanced in gallbladder cancer (GBC) and was involved in the induction of malignant phenotype of GBC. In recent Gallbladder cancer (GBC) is the seventh most common gastro- years, therapeutics that target Hh signaling have focused on intestinal carcinoma and accounts for 1.2% of all cancer cases molecules downstream of smoothened (SMO). The three tran- and 1.7% of all cancer-related deaths (1). GBC develops from scription factors in the Hh signal pathway, glioma-associated metaplasia to dysplasia to carcinoma in situ and then to invasive oncogene homolog 1 (GLI1), GLI2, and GLI3, function down- carcinoma over 5‑15 years (2). During this time, GBC exhibits stream of SMO, but their biological role in GBC remains few characteristic symptoms, and numerous cases have already unclear. In the present study, the biological significance of developed into locally advanced or metastasized cancer by the GLI1, GLI2, and GLI3 were analyzed with the aim of devel- time of diagnosis. Gemcitabine (GEM), cisplatin (CDDP), and oping novel treatments for GBC.
    [Show full text]
  • Distinct Activities of Gli1 and Gli2 in the Absence of Ift88 and the Primary Cilia
    Journal of Developmental Biology Article Distinct Activities of Gli1 and Gli2 in the Absence of Ift88 and the Primary Cilia Yuan Wang 1,2,†, Huiqing Zeng 1,† and Aimin Liu 1,* 1 Department of Biology, Eberly College of Sciences, Center for Cellular Dynamics, Huck Institute of Life Science, The Penn State University, University Park, PA 16802, USA; [email protected] (Y.W.); [email protected] (H.Z.) 2 Department of Occupational Health, School of Public Health, China Medical University, No.77 Puhe Road, Shenyang North New Area, Shenyang 110122, China * Correspondence: [email protected]; Tel.: +1-814-865-7043 † These authors contributed equally to this work. Received: 2 November 2018; Accepted: 16 February 2019; Published: 19 February 2019 Abstract: The primary cilia play essential roles in Hh-dependent Gli2 activation and Gli3 proteolytic processing in mammals. However, the roles of the cilia in Gli1 activation remain unresolved due to the loss of Gli1 transcription in cilia mutant embryos, and the inability to address this question by overexpression in cultured cells. Here, we address the roles of the cilia in Gli1 activation by expressing Gli1 from the Gli2 locus in mouse embryos. We find that the maximal activation of Gli1 depends on the cilia, but partial activation of Gli1 by Smo-mediated Hh signaling exists in the absence of the cilia. Combined with reduced Gli3 repressors, this partial activation of Gli1 leads to dorsal expansion of V3 interneuron and motor neuron domains in the absence of the cilia. Moreover, expressing Gli1 from the Gli2 locus in the presence of reduced Sufu has no recognizable impact on neural tube patterning, suggesting an imbalance between the dosages of Gli and Sufu does not explain the extra Gli1 activity.
    [Show full text]
  • Hedgehog Signalling in the Tumourigenesis and Metastasis of Osteosarcoma, and Its Potential Value in the Clinical Therapy Of
    Yao et al. Cell Death and Disease (2018) 9:701 DOI 10.1038/s41419-018-0647-1 Cell Death & Disease REVIEW ARTICLE Open Access Hedgehog signalling in the tumourigenesis and metastasis of osteosarcoma, and its potential value in the clinical therapy of osteosarcoma Zhihong Yao1,LeiHan1, Yongbin Chen2,FeiHe3,BinSun3, Santosh kamar1, Ya Zhang1,YihaoYang1, Cao Wang1 and Zuozhang Yang1 Abstract The Hedgehog (Hh) signalling pathway is involved in cell differentiation, growth and tissue polarity. This pathway is also involved in the progression and invasion of various human cancers. Osteosarcoma, a subtype of bone cancer, is commonly seen in children and adolescents. Typically, pulmonary osteosarcoma metastases are especially difficult to control. In the present paper, we summarise recent studies on the regulation of osteosarcoma progression and metastasis by downregulating Hh signalling. We also summarise the crosstalk between the Hh pathway and other cancer-related pathways in the tumourigenesis of various cancers. We further summarise and highlight the therapeutic value of potential inhibitors of Hh signalling in the clinical therapy of human cancers. 1234567890():,; 1234567890():,; Facts Open questions ● The Hh pathway regulates the progression of ● How does the Hh pathway regulate the tumourigenic osteosarcoma. progression and invasion of human osteosarcoma? ● The Hh pathway is involved in the metastasis of ● How does the Hh pathway interact with other osteosarcoma into other organs, such as the lungs. cancer-related pathways in the progression and ● The Hh pathway crosstalks with other cancer-related metastasis of cancers? pathways in the tumourigenesis of cancers. ● Could the Hh pathway be used as a target or ● The therapeutic value of the Hh pathway in the biomarker in clinical therapy for human clinical therapy of osteosarcoma is summarised.
    [Show full text]
  • TDP-43 Regulates Retinoblastoma Protein Phosphorylation Through the Repression of Cyclin-Dependent Kinase 6 Expression
    TDP-43 regulates retinoblastoma protein phosphorylation through the repression of cyclin-dependent kinase 6 expression Youhna M. Ayala*, Tom Misteli†, and Francisco E. Baralle*‡ *International Centre for Genetic Engineering and Biotechnology, Padriciano 99, 34012 Trieste, Italy; and †Laboratory of Receptor Biology and Gene Expression, National Cancer Institute, National Institutes of Health, 41 Library Drive, Building 41, Bethesda, MD 20892 Communicated by Jorge E. Allende, University of Chile, Santiago, Chile, January 21, 2008 (received for review September 3, 2007) TDP-43 (for TAR DNA binding protein) is a highly conserved we found that TDP-43 silencing alters cell cycle distribution and heterogeneous nuclear ribonucleoprotein (hnRNP) involved in spe- induces apoptosis. cific pre-mRNA splicing and transcription events. TDP-43 recently has been identified as the main component of cytoplasmic inclu- Results sions in frontotemporal lobar degeneration (FTLD) and amyotro- TDP-43 Down-Regulation Alters the Expression of pRb-Related Fac- phic lateral sclerosis (ALS), two neurodegenerative disorders. The tors. TDP-43 was depleted from HeLa cells by RNAi routinely cellular role of this protein remains to be identified. Here, we show achieving Ͼ90% silencing as measured by Western blot, 48 h that loss of TDP-43 results in dysmorphic nuclear shape, misregu- after small interfering RNA (siRNA) transfection (3, 5). RNA lation of the cell cycle, and apoptosis. Removal of TDP-43 in human microarray analysis was performed on TDP-43 depleted and cells significantly increases cyclin-dependent kinase 6 (Cdk6) pro- control treated cells. The data obtained indicated altered levels tein and transcript levels. The control of Cdk6 expression mediated of several cell proliferation factors in TDP-43-silenced cells.
    [Show full text]