Identification of Small Molecule Inhibitors Targeting Truncated

Total Page:16

File Type:pdf, Size:1020Kb

Identification of Small Molecule Inhibitors Targeting Truncated IDENTIFICATION OF SMALL MOLECULE INHIBITORS TARGETING TRUNCATED ADENOMATOUS POLYPOSIS COLI (APC) AS A NOVEL THERAPEUTIC STRATEGY IN COLORECTAL CANCER APPROVED BY SUPERVISORY COMMITTEE Jerry W. Shay, Ph.D., Mentor Woodring E. Wright, M.D., Ph.D., Mentor Ralph Deberardinis, M.D., Ph.D., Chair Michael Roth, Ph.D. Michael White, Ph.D. ACKNOWLEDGMENTS I would like to express my deepest gratitude to my mentors Dr. Jerry Shay and Dr. Woodring Wright, for their support, encouragement, valuable guidance and instrumental criticism throughout my graduate study. Thank you for sharing your knowledge with me and providing me a strong foundation for my future career as a research scientist. I would like to thank my thesis committee members, Dr. Ralph Deberardinis, Dr. Michael Roth and Dr. Michael White for their invaluable suggestions and constructive feedbacks. I would also like to thank all the current and former lab members of the Shay/Wright lab. In particular, I would like to thank my colleagues Dr. Sang Bum Kim, Dr. Ugur Eskiocak and Dr. Guido Stadler for their experimental guidance and assistance. Likewise, thanks to Sze Wong, Ilgen Mender, Ejun Huang and Gaoxiang Jia for their kind friendship and encouragement. Special thanks to Kevin Kennon for his administrative support. Finally, I would like to thank my parents, Jie Wang and Bin Zhang, and my husband Wenhao Chen for their unconditional support, encouragement and everlasting love. Thank you for being with me every step throughout my life. IDENTIFICATION OF SMALL MOLECULE INHIBITORS TARGETING TRUNCATED ADENOMATOUS POLYPOSIS COLI (APC) AS ANOVEL THERAPEUTIC STRATEGY IN COLORECTAL CANCER By Lu Zhang DISSERTATION Presented to the Faculty of the Graduate School of Biomedical Sciences The University of Texas Southwestern Medical Center at Dallas In Partial Fulfillment of the Requirements For the Degree of Doctor Of DOCTOR OF PHILOSOPHY University Of Texas Southwestern Medical Center Dallas, Texas August, 2014 Copyright by Lu Zhang, 2014 All Rights Reserved IDENTIFICATION OF SMALL MOLECULE INHIBITORS TARGETING TRUNCATED ADENOMATOUS POLYPOSIS COLI (APC) AS A NOVEL THERAPEUTIC STRATEGY IN COLORECTAL CANCER Lu Zhang, Ph.D. The University of Texas Southwestern Medical Center at Dallas, 2014 Jerry W.Shay, Ph.D., Woodring E. Wright, M.D./Ph.D. Abstract Adenomatous polyposis coli (APC) is a tumor suppressor gene that is mutated in the vast majority of colorectal tumors. Inactivation of this gene is a key and early event in colorectal tumorigenesis. APC primarily acts as a negative regulator of Wnt pathway by aiding in degradation of β-catenin. Further studies have suggested that APC plays important roles in several other cellular processes, including cell adhesion and migration, organization of actin and microtubule networks, spindle formation and chromosome segregation. Mutations in APC gene generate truncated gene products, leading to deregulation of these processes. Accumulating evidence suggest that these C terminally truncated APC may have gain of function properties beyond their loss of tumor suppressive function. Both the gain and v vi loss of function of APC truncations contribute to CRC initiation and progression. Utilizing a series of isogenic immortalized human colonic epithelial cells (HCECs), we have screened a 200K compound library for small molecules that exhibited selective cytotoxicity towards HCECs expressing truncated APC. We identified a compound, TASIN-1(Truncated APC Selective Inhibitor-1), which showed selective cytotoxicity towards HCECs and colorectal cancer (CRC) cells with APC truncations. TASIN-1 induces apoptotic cell death in APC truncated cells and its effect is independent of canonical Wnt pathway activity. Short hairpin RNA (shRNA) mediated knockdown of truncated APC confers resistance to TASIN-1. Additionally, TASIN-1 can interact with in vitro translated APC fragments, implicating truncated APC in the mechanism of action of TASIN-1. TASIN-1 inhibits tumor growth in both xenograft and genetic CRC mouse models. TASIN-1 may represent a novel therapeutic strategy for colon cancer prevention and intervention. Table of Contents Page Abstract ......................................................................................................................................................... v Prior Publications ......................................................................................................................................... vi List of Figures ............................................................................................................................................. vii List of Tables ............................................................................................................................................... ix Chapter 1 Introduction .................................................................................................................................. 1 Colorectal Cancer (CRC) .......................................................................................................................... 1 Adenomatous polyposis coli (APC) .......................................................................................................... 6 The APC gene ....................................................................................................................................... 6 APC Protein and its Many Functions .................................................................................................... 7 APC Mutation in Sporadic CRC ......................................................................................................... 11 Loss of Tumor Suppressive Function of APC in CRC ....................................................................... 12 Dominant-negative Effects of APC .................................................................................................... 15 APC Truncation: Switch from Tumor Suppressor to Oncogene? ....................................................... 16 Lessons from genetic mouse models ...................................................................................................... 20 Overview of chemical genetics-based target identification in drug discovery ...... Error! Bookmark not defined. Target-based versus chemical genetics drug discovery approaches ................................................... 22 Chemical proteomics for protein target identification in chemical genetics drug discovery .............. 23 Additional approaches for target identification-transcriptional profiling ........................................... 26 Additional approaches for target identification-genetics screening .................................................... 27 Purpose and significance ......................................................................................................................... 28 Chapter 2 . Cell Survival Pathways in Colorectal Cancer (CRC) Cells: Targeting Mutant Adenomatous Polyposis Coli (APC) .................................................................................................................................. 36 Introduction ............................................................................................................................................. 36 Materials and Methods ............................................................................................................................ 37 Results ..................................................................................................................................................... 43 Discussion ............................................................................................................................................... 48 Chapter 3 . Identification of Novel Driver Tumor Suppressors through Functional Interrogation of Putative Passenger Mutations in Colorectal Cancer ................................................................................... 68 Introduction ............................................................................................................................................. 69 Materials and Methods ............................................................................................................................ 71 iv v Results ..................................................................................................................................................... 72 Discussion ............................................................................................................................................... 76 Chapter 4 . Exome Sequencing of Normal and Isogenic Transformed Human Colonic Epithelial Cells (HCECs) Reveals Novel Genes Potentially Involved in the Early Stages of Colorectal Tumorigenesis ... 96 Introduction ............................................................................................................................................. 96 Materials and Methods ............................................................................................................................ 98 Results ..................................................................................................................................................... 99 Discussion ............................................................................................................................................. 103 Chapter 5 . Conclusions and Future Perspectives ....................................................................................
Recommended publications
  • XPO1E571K Mutation Modifies Exportin 1 Localisation And
    cancers Article XPO1E571K Mutation Modifies Exportin 1 Localisation and Interactome in B-Cell Lymphoma Hadjer Miloudi 1, Élodie Bohers 1,2, François Guillonneau 3 , Antoine Taly 4,5 , Vincent Cabaud Gibouin 6,7 , Pierre-Julien Viailly 1,2 , Gaëtan Jego 6,7 , Luca Grumolato 8 , Fabrice Jardin 1,2 and Brigitte Sola 1,* 1 INSERM U1245, Unicaen, Normandie University, F-14000 Caen, France; [email protected] (H.M.); [email protected] (E.B.); [email protected] (P.-J.V.); [email protected] (F.J.) 2 Centre de lutte contre le Cancer Henri Becquerel, F-76000 Rouen, France 3 Plateforme Protéomique 3P5, Université de Paris, Institut Cochin, INSERM, CNRS, F-75014 Paris, France; [email protected] 4 Laboratoire de Biochimie Théorique, CNRS UPR 9030, Université de Paris, F-75005 Paris, France; [email protected] 5 Institut de Biologie Physico-Chimique, Fondation Edmond de Rothschild, PSL Research University, F-75005 Paris, France 6 INSERM, LNC UMR1231, F-21000 Dijon, France; [email protected] (V.C.G.); [email protected] (G.J.) 7 Team HSP-Pathies, University of Burgundy and Franche-Comtée, F-21000 Dijon, France 8 INSERM U1239, Unirouen, Normandie University, F-76130 Mont-Saint-Aignan, France; [email protected] * Correspondence: [email protected]; Tel.: +33-2-3156-8210 Received: 11 September 2020; Accepted: 28 September 2020; Published: 30 September 2020 Simple Summary: Almost 25% of patients with either primary mediastinal B-cell lymphoma (PMBL) or classical Hodgkin lymphoma (cHL) possess a recurrent mutation of the XPO1 gene encoding the major nuclear export protein.
    [Show full text]
  • Cyclin D1/Cyclin-Dependent Kinase 4 Interacts with Filamin a and Affects the Migration and Invasion Potential of Breast Cancer Cells
    Published OnlineFirst February 28, 2010; DOI: 10.1158/0008-5472.CAN-08-1108 Tumor and Stem Cell Biology Cancer Research Cyclin D1/Cyclin-Dependent Kinase 4 Interacts with Filamin A and Affects the Migration and Invasion Potential of Breast Cancer Cells Zhijiu Zhong, Wen-Shuz Yeow, Chunhua Zou, Richard Wassell, Chenguang Wang, Richard G. Pestell, Judy N. Quong, and Andrew A. Quong Abstract Cyclin D1 belongs to a family of proteins that regulate progression through the G1-S phase of the cell cycle by binding to cyclin-dependent kinase (cdk)-4 to phosphorylate the retinoblastoma protein and release E2F transcription factors for progression through cell cycle. Several cancers, including breast, colon, and prostate, overexpress the cyclin D1 gene. However, the correlation of cyclin D1 overexpression with E2F target gene regulation or of cdk-dependent cyclin D1 activity with tumor development has not been identified. This suggests that the role of cyclin D1 in oncogenesis may be independent of its function as a cell cycle regulator. One such function is the role of cyclin D1 in cell adhesion and motility. Filamin A (FLNa), a member of the actin-binding filamin protein family, regulates signaling events involved in cell motility and invasion. FLNa has also been associated with a variety of cancers including lung cancer, prostate cancer, melanoma, human bladder cancer, and neuroblastoma. We hypothesized that elevated cyclin D1 facilitates motility in the invasive MDA-MB-231 breast cancer cell line. We show that MDA-MB-231 motility is affected by disturbing cyclin D1 levels or cyclin D1-cdk4/6 kinase activity.
    [Show full text]
  • Constitutive Scaffolding of Multiple Wnt Enhanceosome Components By
    RESEARCH ARTICLE Constitutive scaffolding of multiple Wnt enhanceosome components by Legless/ BCL9 Laurens M van Tienen, Juliusz Mieszczanek, Marc Fiedler, Trevor J Rutherford, Mariann Bienz* MRC Laboratory of Molecular Biology, Cambridge, United Kingdom Abstract Wnt/b-catenin signaling elicits context-dependent transcription switches that determine normal development and oncogenesis. These are mediated by the Wnt enhanceosome, a multiprotein complex binding to the Pygo chromatin reader and acting through TCF/LEF- responsive enhancers. Pygo renders this complex Wnt-responsive, by capturing b-catenin via the Legless/BCL9 adaptor. We used CRISPR/Cas9 genome engineering of Drosophila legless (lgs) and human BCL9 and B9L to show that the C-terminus downstream of their adaptor elements is crucial for Wnt responses. BioID proximity labeling revealed that BCL9 and B9L, like PYGO2, are constitutive components of the Wnt enhanceosome. Wnt-dependent docking of b-catenin to the enhanceosome apparently causes a rearrangement that apposes the BCL9/B9L C-terminus to TCF. This C-terminus binds to the Groucho/TLE co-repressor, and also to the Chip/LDB1-SSDP enhanceosome core complex via an evolutionary conserved element. An unexpected link between BCL9/B9L, PYGO2 and nuclear co-receptor complexes suggests that these b-catenin co-factors may coordinate Wnt and nuclear hormone responses. DOI: 10.7554/eLife.20882.001 *For correspondence: mb2@mrc- Introduction lmb.cam.ac.uk The Wnt/b-catenin signaling cascade is an ancient cell communication pathway that operates con- Competing interests: The text-dependent transcriptional switches to control animal development and tissue homeostasis authors declare that no (Cadigan and Nusse, 1997).
    [Show full text]
  • 1325.Full-Text.Pdf
    IN THIS ISSUE APC Mutation Position Dictates Effect of Tankyrase Inhibition in Colorectal Cancer • The effects of APC mutations, which in- • New animal models, human cells, and or- • Cases with different mutations in crease WNT signaling in colorectal can- ganoids were used to circumvent issues the same gene should be evaluated cer, can be reversed by TNKS inhibition . with mouse colorectal cancer models . separately for therapeutic response . Hyperactive WNT signaling is tumor growth in vivo. However, whether TNKS inhibition seen in most colorectal cancers, was effective depended on the mechanism of APC disrup- and inactivating mutations in the tion: APC mutants with truncations in the mutation cluster tumor suppressor adenomatous region were still able to regulate β-catenin and responded to polyposis coli (APC)—a scaffold TNKS blockade, whereas this was not the case when there protein mediating the formation were truncations earlier in the sequence. Truncations in the of the destruction complex (DC) mutation cluster region are commonly observed in patients, that facilitates β-catenin degrada- whereas the earlier truncations are present in commonly used tion—is the cause in 80% of such mouse models. Collectively, these results indicate that TNKS cases. Restoring DC activity (and, inhibition can restore control of WNT signaling in some thus, normal WNT signaling) in the context of inactivated APC-mutant cases and illustrate that different mutations in APC is possible through pharmacologic inhibition of tanky- the same gene, even those causing the same phenotype (in rase (TNKS) 1 and 2, which are functionally redundant. Using this case, WNT hyperactivation), can respond differently to APC-mutant animal models, human cells, and ex vivo orga- targeted therapies.
    [Show full text]
  • Supplementary Figures
    Mena regulates the LINC complex to control actin–nuclear lamina associations, trans-nuclear membrane signalling and cancer gene expression Frederic Li Mow Chee!, Bruno Beernaert!, Alexander Loftus!, Yatendra Kumar", Billie G. C. Griffith!, Jimi C. Wills!, Ann P. Wheeler#, J. Douglas Armstrong$, Maddy Parsons%, Irene M. Leigh,(, Charlotte M. Proby&, Alex von Kriegsheim!, Wendy A. Bickmore", Margaret C. Frame,* & Adam Byron,* Supplementary Information Supplementary Figure 1 Supplementary Figure 2 Supplementary Figure 3 Supplementary Table 1 Supplementary Table 2 Supplementary Table 3 Supplementary Table 4 !Cancer Research UK Edinburgh Centre, Institute of Genetics and Cancer, University of Edinburgh, Edinburgh EH< =XR, UK. "MRC Human Genetics Unit, Institute of Genetics and Cancer, University of Edinburgh, Edinburgh EH< =XU, UK. #Advanced Imaging Resource, Institute of Genetics and Cancer, University of Edinburgh, Edinburgh EH< =XU, UK. $Simons Initiative for the Developing Brain, School of Informatics, University of Edinburgh, Edinburgh EHH IYL, UK. %Randall Centre for Cell and Molecular Biophysics, King’s College London, London SEM MUL, UK. &Division of Molecular and Clinical Medicine, School of Medicine, University of Dundee, Dundee DD <HN, UK. 'Institute of Dentistry, Barts and the London School of Medicine and Dentistry, Queen Mary University of London, London EM =AT, UK. *email: [email protected] or [email protected] 1 a cSCC IAC correlation b cSCC IAC pathways c Core adhesome network ENAH −log10(q) MACF1 CSRP1 Met1 Met4 0 5 10 + + CORO2A Integrin signalling + CFL1 pathway PRNP ILK + HSPB1 PALLD PPFIA1 TES RDX Cytoskeletal regulation + VASP + + ARPC2 by Rho GTPase PPP2CA + Met1 + LASP1 MYH9 + VIM TUBA4A Huntington ITGA3 + disease ITGB4 VCL CAV1 ACTB ROCK1 KTN1 FLNA+ CALR DNA FBLIM1 CORO1B RAC1 + replication +ACTN1 ITGA6 + Met4 ITGAV Parkinson ITGB1 disease Actin cytoskel.
    [Show full text]
  • PSMB5 Antibody Cat
    PSMB5 Antibody Cat. No.: 57-791 PSMB5 Antibody Western blot analysis of PSMB5 using rabbit polyclonal PSMB5 Antibody immunohistochemistry analysis in PSMB5 Antibody using 293 cell lysates (2 ug/lane) either formalin fixed and paraffin embedded human skin tissue nontransfected (Lane 1) or transiently transfected (Lane 2) followed by peroxidase conjugation of the secondary with the PSMB5 gene. antibody and DAB staining. Specifications HOST SPECIES: Rabbit SPECIES REACTIVITY: Human This PSMB5 antibody is generated from rabbits immunized with a KLH conjugated IMMUNOGEN: synthetic peptide between 235-263 amino acids from the C-terminal region of human PSMB5. TESTED APPLICATIONS: IHC-P, WB For WB starting dilution is: 1:1000 APPLICATIONS: For IHC-P starting dilution is: 1:10~50 October 1, 2021 1 https://www.prosci-inc.com/psmb5-antibody-57-791.html PREDICTED MOLECULAR 28 kDa WEIGHT: Properties This antibody is purified through a protein A column, followed by peptide affinity PURIFICATION: purification. CLONALITY: Polyclonal ISOTYPE: Rabbit Ig CONJUGATE: Unconjugated PHYSICAL STATE: Liquid BUFFER: Supplied in PBS with 0.09% (W/V) sodium azide. CONCENTRATION: batch dependent Store at 4˚C for three months and -20˚C, stable for up to one year. As with all antibodies STORAGE CONDITIONS: care should be taken to avoid repeated freeze thaw cycles. Antibodies should not be exposed to prolonged high temperatures. Additional Info OFFICIAL SYMBOL: PSMB5 Proteasome subunit beta type-5, Macropain epsilon chain, Multicatalytic endopeptidase ALTERNATE NAMES: complex epsilon chain, Proteasome chain 6, Proteasome epsilon chain, Proteasome subunit MB1, Proteasome subunit X, PSMB5, LMPX, MB1, X ACCESSION NO.: P28074 PROTEIN GI NO.: 187608890 GENE ID: 5693 USER NOTE: Optimal dilutions for each application to be determined by the researcher.
    [Show full text]
  • Myosin Myth4-FERM Structures Highlight Important Principles of Convergent Evolution
    Myosin MyTH4-FERM structures highlight important principles of convergent evolution Vicente José Planelles-Herreroa,b, Florian Blanca,c, Serena Sirigua, Helena Sirkiaa, Jeffrey Clausea, Yannick Souriguesa, Daniel O. Johnsrudd, Beatrice Amiguesa, Marco Cecchinic, Susan P. Gilberte, Anne Houdussea,1,2, and Margaret A. Titusd,1,2 aStructural Motility, Institut Curie, CNRS, UMR 144, PSL Research University, F-75005 Paris, France; bUPMC Université de Paris 6, Institut de Formation Doctorale, Sorbonne Universités, 75252 Paris Cedex 05, France; cLaboratoire d’Ingénierie des Fonctions Moléculaires, Institut de Science et d’Ingénierie Supramoléculaires, UMR 7006 CNRS, Université de Strasbourg, F-67083 Strasbourg Cedex, France; dDepartment of Genetics, Cell Biology and Development, University of Minnesota, Minneapolis, MN 55455; and eDepartment of Biological Sciences, Rensselaer Polytechnic Institute, Troy, NY 12180 Edited by James A. Spudich, Stanford University School of Medicine, Stanford, CA, and approved March 31, 2016 (received for review January 15, 2016) Myosins containing MyTH4-FERM (myosin tail homology 4-band (Fig. 1). These MF myosins are widespread and likely quite an- 4.1, ezrin, radixin, moesin, or MF) domains in their tails are found cient because they are found in many different branches of the in a wide range of phylogenetically divergent organisms, such as phylogenetic tree (5, 6), including Opisthokonts (which includes humans and the social amoeba Dictyostelium (Dd). Interestingly, Metazoa, unicellular Holozoa, and Fungi), Amoebozoa, and the evolutionarily distant MF myosins have similar roles in the exten- SAR (Stramenopiles, Alveolates, and Rhizaria) (Fig. 1 A and B). sion of actin-filled membrane protrusions such as filopodia and Over the course of hundreds of millions years of parallel evolution bind to microtubules (MT), suggesting that the core functions of the MF myosins have acquired or maintained roles in the formation these MF myosins have been highly conserved over evolution.
    [Show full text]
  • Inhibition of the Nuclear Export Receptor XPO1 As a Therapeutic Target for Platinum-Resistant Ovarian Cancer Ying Chen1, Sandra Catalina Camacho1, Thomas R
    Published OnlineFirst September 20, 2016; DOI: 10.1158/1078-0432.CCR-16-1333 Cancer Therapy: Preclinical Clinical Cancer Research Inhibition of the Nuclear Export Receptor XPO1 as a Therapeutic Target for Platinum-Resistant Ovarian Cancer Ying Chen1, Sandra Catalina Camacho1, Thomas R. Silvers1, Albiruni R.A. Razak2, Nashat Y. Gabrail3, John F. Gerecitano4, Eva Kalir1, Elena Pereira5, Brad R. Evans1, Susan J. Ramus6, Fei Huang1, Nolan Priedigkeit1, Estefania Rodriguez1, Michael Donovan7, Faisal Khan7, Tamara Kalir7, Robert Sebra1, Andrew Uzilov1, Rong Chen1, Rileen Sinha1, Richard Halpert8, Jean-Noel Billaud8, Sharon Shacham9, Dilara McCauley9, Yosef Landesman9, Tami Rashal9, Michael Kauffman9, Mansoor R. Mirza9, Morten Mau-Sørensen10, Peter Dottino5, and John A. Martignetti1,5,11 Abstract Purpose: The high fatality-to-case ratio of ovarian cancer is Results: XPO1 RNA overexpression and protein nuclear directly related to platinum resistance. Exportin-1 (XPO1) is a localization were correlated with decreased survival and plat- nuclear exporter that mediates nuclear export of multiple tumor inum resistance in ovarian cancer. Targeted XPO1 inhibition suppressors. We investigated possible clinicopathologic correla- decreased cell viability and synergistically restored platinum tions of XPO1 expression levels and evaluated the efficacy of sensitivity in both immortalized ovarian cancer cells and XPO1 inhibition as a therapeutic strategy in platinum-sensitive PDCL. The XPO1 inhibitor–mediated apoptosis occurred and -resistant ovarian cancer. through both p53-dependent and p53-independent signaling Experimental Design: XPO1 expression levels were analyzed to pathways. Selinexor treatment, alone and in combination with define clinicopathologic correlates using both TCGA/GEO data- platinum, markedly decreased tumor growth and prolonged sets and tissue microarrays (TMA).
    [Show full text]
  • Defining Functional Interactions During Biogenesis of Epithelial Junctions
    ARTICLE Received 11 Dec 2015 | Accepted 13 Oct 2016 | Published 6 Dec 2016 | Updated 5 Jan 2017 DOI: 10.1038/ncomms13542 OPEN Defining functional interactions during biogenesis of epithelial junctions J.C. Erasmus1,*, S. Bruche1,*,w, L. Pizarro1,2,*, N. Maimari1,3,*, T. Poggioli1,w, C. Tomlinson4,J.Lees5, I. Zalivina1,w, A. Wheeler1,w, A. Alberts6, A. Russo2 & V.M.M. Braga1 In spite of extensive recent progress, a comprehensive understanding of how actin cytoskeleton remodelling supports stable junctions remains to be established. Here we design a platform that integrates actin functions with optimized phenotypic clustering and identify new cytoskeletal proteins, their functional hierarchy and pathways that modulate E-cadherin adhesion. Depletion of EEF1A, an actin bundling protein, increases E-cadherin levels at junctions without a corresponding reinforcement of cell–cell contacts. This unexpected result reflects a more dynamic and mobile junctional actin in EEF1A-depleted cells. A partner for EEF1A in cadherin contact maintenance is the formin DIAPH2, which interacts with EEF1A. In contrast, depletion of either the endocytic regulator TRIP10 or the Rho GTPase activator VAV2 reduces E-cadherin levels at junctions. TRIP10 binds to and requires VAV2 function for its junctional localization. Overall, we present new conceptual insights on junction stabilization, which integrate known and novel pathways with impact for epithelial morphogenesis, homeostasis and diseases. 1 National Heart and Lung Institute, Faculty of Medicine, Imperial College London, London SW7 2AZ, UK. 2 Computing Department, Imperial College London, London SW7 2AZ, UK. 3 Bioengineering Department, Faculty of Engineering, Imperial College London, London SW7 2AZ, UK. 4 Department of Surgery & Cancer, Faculty of Medicine, Imperial College London, London SW7 2AZ, UK.
    [Show full text]
  • Circmyo10 Promotes Osteosarcoma Progression by Regulating Mir-370
    Chen et al. Molecular Cancer (2019) 18:150 https://doi.org/10.1186/s12943-019-1076-1 RESEARCH Open Access CircMYO10 promotes osteosarcoma progression by regulating miR-370-3p/ RUVBL1 axis to enhance the transcriptional activity of β-catenin/LEF1 complex via effects on chromatin remodeling Junxin Chen1†, Gang Liu1†, Yizheng Wu1†, Jianjun Ma1†, Hongfei Wu2, Ziang Xie1, Shuai Chen1, Yute Yang1, Shengyu Wang1, Panyang Shen1, Yifan Fang3, Shunwu Fan1, Shuying Shen1* and Xiangqian Fang1* Abstract Background: CircMYO10 is a circular RNA generated by back-splicing of gene MYO10 and is upregulated in osteosarcoma cell lines, but its functional role in osteosarcoma is still unknown. This study aimed to clarify the mechanism of circMYO10 in osteosarcoma. Methods: CircMYO10 expression in 10 paired osteosarcoma and chondroma tissues was assessed by quantitative reverse transcription polymerase chain reaction (PCR). The function of circMYO10/miR-370-3p/RUVBL1 axis was assessed regarding two key characteristics: proliferation and endothelial–mesenchymal transition (EMT). Bioinformatics analysis, western blotting, real-time PCR, fluorescence in situ hybridization, immunoprecipitation, RNA pull-down assays, luciferase reporter assays, chromatin immunoprecipitation, and rescue experiments were used to evaluate the mechanism. Stably transfected MG63 cells were injected via tail vein or subcutaneously into nude mice to assess the role of circMYO10 in vivo. Results: CircMYO10 was significantly upregulated, while miR-370-3p was downregulated, in osteosarcoma cell lines and human osteosarcoma samples. Silencing circMYO10 inhibited cell proliferation and EMT in vivo and in vitro. Mechanistic investigations revealed that miR-370-3p targets RUVBL1 directly, and inhibits the interaction between RUVBL1 and β-catenin/LEF1 complex while circMYO10 showed a contrary effect via the inhibition of miR-370-3p.
    [Show full text]
  • Vascular and Connective Tissue Anomalies Associated with X-Linked Periventricular Heterotopia Due to Mutations in Filamin A
    European Journal of Human Genetics (2013) 21, 494–502 & 2013 Macmillan Publishers Limited All rights reserved 1018-4813/13 www.nature.com/ejhg ARTICLE Vascular and connective tissue anomalies associated with X-linked periventricular heterotopia due to mutations in Filamin A Eyal Reinstein*,1, Sophia Frentz2, Tim Morgan2, Sixto Garcı´a-Min˜au´r3, Richard J Leventer4, George McGillivray5, Mitchel Pariani1, Anthony van der Steen6, Michael Pope6, Muriel Holder-Espinasse7, Richard Scott8,9, Elizabeth M Thompson10, Terry Robertson11, Brian Coppin12, Robert Siegel13, Montserrat Bret Zurita14, Jose I Rodrı´guez15, Carmen Morales15, Yuri Rodrigues15, Joaquı´n Arcas16, Anand Saggar17, Margaret Horton18, Elaine Zackai18, John M Graham1, David L Rimoin1,{ and Stephen P Robertson2 Mutations conferring loss of function at the FLNA (encoding filamin A) locus lead to X-linked periventricular nodular heterotopia (XL-PH), with seizures constituting the most common clinical manifestation of this disorder in female heterozygotes. Vascular dilatation (mainly the aorta), joint hypermobility and variable skin findings are also associated anomalies, with some reports suggesting that this might represents a separate syndrome allelic to XL-PH, termed as Ehlers-Danlos syndrome-periventricular heterotopia variant (EDS-PH). Here, we report a cohort of 11 males and females with both hypomorphic and null mutations in FLNA that manifest a wide spectrum of connective tissue and vascular anomalies. The spectrum of cutaneous defects was broader than previously described and is inconsistent with a specific type of EDS. We also extend the range of vascular anomalies associated with XL-PH to included peripheral arterial dilatation and atresia. Based on these observations, we suggest that there is little molecular or clinical justification for considering EDS-PH as a separate entity from XL-PH, but instead propose that there is a spectrum of vascular and connective tissues anomalies associated with this condition for which all individuals with loss-of-function mutations in FLNA should be evaluated.
    [Show full text]
  • Human Periprostatic Adipose Tissue: Secretome from Patients With
    CANCER GENOMICS & PROTEOMICS 16 : 29-58 (2019) doi:10.21873/cgp.20110 Human Periprostatic Adipose Tissue: Secretome from Patients With Prostate Cancer or Benign Prostate Hyperplasia PAULA ALEJANDRA SACCA 1, OSVALDO NÉSTOR MAZZA 2, CARLOS SCORTICATI 2, GONZALO VITAGLIANO 3, GABRIEL CASAS 4 and JUAN CARLOS CALVO 1,5 1Institute of Biology and Experimental Medicine (IBYME), CONICET, Buenos Aires, Argentina; 2Department of Urology, School of Medicine, University of Buenos Aires, Clínical Hospital “José de San Martín”, Buenos Aires, Argentina; 3Department of Urology, Deutsches Hospital, Buenos Aires, Argentina; 4Department of Pathology, Deutsches Hospital, Buenos Aires, Argentina; 5Department of Biological Chemistry, School of Exact and Natural Sciences, University of Buenos Aires, Buenos Aires, Argentina Abstract. Background/Aim: Periprostatic adipose tissue Prostate cancer (PCa) is the second most common cancer in (PPAT) directs tumour behaviour. Microenvironment secretome men worldwide. While most men have indolent disease, provides information related to its biology. This study was which can be treated properly, the problem consists in performed to identify secreted proteins by PPAT, from both reliably distinguishing between indolent and aggressive prostate cancer and benign prostate hyperplasia (BPH) disease. Evidence shows that the microenvironment affects patients. Patients and Methods: Liquid chromatography-mass tumour behavior. spectrometry-based proteomic analysis was performed in Adipose tissue microenvironment is now known to direct PPAT-conditioned media (CM) from patients with prostate tumour growth, invasion and metastases (1, 2). Adipose cancer (CMs-T) (stage T3: CM-T3, stage T2: CM-T2) or tissue is adjacent to the prostate gland and the site of benign disease (CM-BPH). Results: The highest number and invasion of PCa.
    [Show full text]