DOCSLIB.ORG

Functions of Drosophila Pak (P21-Activated Kinase) in Morphogenesis: a Mechanistic Model Based on Cellular, Molecular, and Genetic Studies

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Functions of Drosophila Pak (P21-Activated Kinase) in Morphogenesis: a Mechanistic Model Based on Cellular, Molecular, and Genetic Studies Functions of Drosophila Pak (p21-activated kinase) in Morphogenesis: A Mechanistic Model based on Cellular, Molecular, and Genetic Studies Item Type text; Electronic Dissertation Authors Lewis, Sara Ann Publisher The University of Arizona. Rights Copyright © is held by the author. Digital access to this material is made possible by the University Libraries, University of Arizona. Further transmission, reproduction or presentation (such as public display or performance) of protected items is prohibited except with permission of the author. Download date 07/10/2021 10:07:41 Link to Item http://hdl.handle.net/10150/594389 FUNCTIONS OF DROSOPHILA PAK (P21-ACTIVATED KINASE) IN MORPHOGENESIS: A MECHANISTIC MODEL BASED ON CELLULAR, MOLECULAR, AND GENETIC STUDIES by Sara Lewis A Dissertation Submitted to the Faculty of the GRADUATE INTERDISCIPLINARY PROGRAM in Partial Fulfillment of the Requirements for the Degree of DOCTOR OF PHILOSOPHY IN NEUROSCIENCE In the Graduate College THE UNIVERSITY OF ARIZONA 2015 THE UNIVERSITY OF ARIZONA GRADUATE COLLEGE As members of the Dissertation Committee, we certify that we have read the dissertation prepared by Sara Lewis, titled “Functions of Drosophila Pak (p21-activated kinase) in morphogenesis: a mechanistic model based on cellular, molecular and genetic studies” and recommend that it be accepted as fulfilling the dissertation requirement for the Degree of Doctor of Philosophy. _______________________________________________________________________ Date: November 18, 2015 Dr. Linda Restifo _______________________________________________________________________ Date: November 18, 2015 Dr. Jean Wilson _______________________________________________________________________ Date: November 18, 2015 Dr. Konrad Zinsmaier _______________________________________________________________________ Date: November 18, 2015 Dr. Lynne Oland Final approval and acceptance of this dissertation is contingent upon the candidate’s submission of the final copies of the dissertation to the Graduate College. I hereby certify that I have read this dissertation prepared under my direction and recommend that it be accepted as fulfilling the dissertation requirement. ________________________________________________ Date: November 18, 2015 Dissertation Director: Dr. Alan Nighorn 2 Statement by Author This dissertation has been submitted in partial fulfillment of the requirements for an advanced degree at the University of Arizona and is deposited in the University Library to be made available to borrowers under rules of the Library. Brief quotations from this dissertation are allowable without special permission, provided that an accurate acknowledgement of the source is made. Requests for permission for extended quotation from or reproduction of this manuscript in whole or in part may be granted by the head of the major department or the Dean of the Graduate College when in his or her judgment the proposed use of the material is in the interests of scholarship. In all other instances, however, permission must be obtained from the author. ________________________________________________ SIGNED: Sara Lewis 3 Acknowledgements I am grateful to numerous undergraduates and other students who assisted with this research, both directly and indirectly, over the years: Monica Chaung, Adrian Ostertag, Aubrey Downey, Allegra Cohen, Cori Schwartzlow, Cecilia Brown, Rachel Doser, and Grace Samtani. I especially appreciate the hard work of Lauren O’Neill and Josh Paree in immunostaining, morphological analysis, NeuronMetrics, and fly rearing. Many thanks to Judith Tello for years of support as my peer and friend. Thank you to Jennifer Tifft and William Lewis for editing help on this thesis. I would like to express my appreciation to Anna Burns for tissue culture training and Patty Jansma for outstanding microscopy assistance and knowledge. I am forever indebted to the expertise and mentoring of Dave Andrew and Robert Kraft. Thank you for all of your training, feedback, and support to help me develop as a scientist. I would like to thank the members of my committee: Lynne Oland, Jean Wilson, Konrad Zinsmaier, and Alan Nighorn. I appreciate your advice and the numerous ways you supported my research over the years. I am also grateful to the GIDP in Neuroscience for funding and support and to all of the students and faculty who made graduate school such a rich learning opportunity. Finally, I would like to thank my mentor, Linda Restifo, for constantly challenging me. I am grateful to my husband, William Lewis, for constant support and encouragement. 4 Table of Contents List of Tables and Figures ............................................................................................................................. 7 Abstract ......................................................................................................................................................... 8 List of abbreviations ................................................................................................................................... 10 Chapter I: Introduction ................................................................................................................................ 11 Morphogenesis is the genetically-regulated process of changing tissue shape to form body structures ............................................................................................................................... 11 Intellectual disability, microcephaly, and PAK3 ............................................................................ 13 PAK3 regulates proteins that stabilize the actin cytoskeleton ....................................................... 16 Rodent and Drosophila models of PAK3 ....................................................................................... 19 Drosophila mushroom bodies are higher-order brain structures whose morphology is genetically regulated ..................................................................................................................... 23 Non-muscle myosins have diverse, critical roles in morphology and nervous system physiology ...................................................................................................................................... 25 Statement of problem addressed in dissertation ............................................................................. 27 Chapter II: Selective regulation of appendage and mushroom body morphogenesis by Drosophila Pak (p21-activated kinase) ................................................................................................................................. 31 Abstract .......................................................................................................................................... 31 Introduction .................................................................................................................................... 32 5 Results ............................................................................................................................................ 35 Discussion ...................................................................................................................................... 42 Materials and Methods ................................................................................................................... 48 Figures and Legends ...................................................................................................................... 54 Chapter III: Pak (p21-activated kinase) regulates neurite arbor size and brain structure morphology with apparent roles in branch stability and inhibition of myosin regulatory light chain phosphorylation .......... 68 Abstract .......................................................................................................................................... 68 Introduction .................................................................................................................................... 69 Results ............................................................................................................................................ 72 Discussion ...................................................................................................................................... 79 Materials and Methods ................................................................................................................... 83 Figures and Legends ...................................................................................................................... 89 Chapter IV: Summary and Future directions ............................................................................................ 101 Appendix A: Cofilin (twinstar) and Pak interact to regulate neurite branches, but not total neurite length in vitro ....................................................................................................................................................... 106 References ................................................................................................................................................. 110 6 List of Tables and Figures Table 1: Human mutations in PAK3, protein products and function ......................................................... 29 Table 2: Antibodies used for immunolabeling ........................................................................................... 49 Fig. 1: Pak molecular signaling
Load more

Recommended publications
  • Viewer 4.0 Software [73]
    BMC Genomics BioMed Central Research Open Access Bioinformatic search of plant microtubule-and cell cycle related serine-threonine protein kinases Pavel A Karpov1, Elena S Nadezhdina2,3,AllaIYemets1, Vadym G Matusov1, Alexey Yu Nyporko1,NadezhdaYuShashina3 and Yaroslav B Blume*1 Addresses: 1Institute of Food Biotechnology and Genomics, National Academy of Sciences of Ukraine, 04123 Kyiv, Ukraine, 2Institute of Protein Research, Russian Academy of Sciences, 142290 Pushchino, Moscow Region, Russian Federation and 3AN Belozersky Institute of Physical- Chemical Biology, Moscow State University, Leninsky Gory, 119992 Moscow, Russian Federation E-mail: Pavel A Karpov - [email protected]; Elena S Nadezhdina - [email protected]; Alla I Yemets - [email protected]; Vadym G Matusov - [email protected]; Alexey Yu Nyporko - [email protected]; Nadezhda Yu Shashina - [email protected]; Yaroslav B Blume* - [email protected] *Corresponding author from International Workshop on Computational Systems Biology Approaches to Analysis of Genome Complexity and Regulatory Gene Networks Singapore 20-25 November 2008 Published: 10 February 2010 BMC Genomics 2010, 11(Suppl 1):S14 doi: 10.1186/1471-2164-11-S1-S14 This article is available from: http://www.biomedcentral.com/1471-2164/11/S1/S14 Publication of this supplement was made possible with help from the Bioinformatics Agency for Science, Technology and Research of Singapore and the Institute for Mathematical Sciences at the National University of Singapore. © 2010 Karpov et al; licensee BioMed Central Ltd. This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
    [Show full text]
  • Endocrine System Local Gene Expression
    Copyright 2008 By Nathan G. Salomonis ii Acknowledgments Publication Reprints The text in chapter 2 of this dissertation contains a reprint of materials as it appears in: Salomonis N, Hanspers K, Zambon AC, Vranizan K, Lawlor SC, Dahlquist KD, Doniger SW, Stuart J, Conklin BR, Pico AR. GenMAPP 2: new features and resources for pathway analysis. BMC Bioinformatics. 2007 Jun 24;8:218. The co-authors listed in this publication co-wrote the manuscript (AP and KH) and provided critical feedback (see detailed contributions at the end of chapter 2). The text in chapter 3 of this dissertation contains a reprint of materials as it appears in: Salomonis N, Cotte N, Zambon AC, Pollard KS, Vranizan K, Doniger SW, Dolganov G, Conklin BR. Identifying genetic networks underlying myometrial transition to labor. Genome Biol. 2005;6(2):R12. Epub 2005 Jan 28. The co-authors listed in this publication developed the hierarchical clustering method (KP), co-designed the study (NC, AZ, BC), provided statistical guidance (KV), co- contributed to GenMAPP 2.0 (SD) and performed quantitative mRNA analyses (GD). The text of this dissertation contains a reproduction of a figure from: Yeo G, Holste D, Kreiman G, Burge CB. Variation in alternative splicing across human tissues. Genome Biol. 2004;5(10):R74. Epub 2004 Sep 13. The reproduction was taken without permission (chapter 1), figure 1.3. iii Personal Acknowledgments The achievements of this doctoral degree are to a large degree possible due to the contribution, feedback and support of many individuals. To all of you that helped, I am extremely grateful for your support.
    [Show full text]
  • Developmental Biology 322 (2008) 95–108
    Developmental Biology 322 (2008) 95–108 Contents lists available at ScienceDirect Developmental Biology journal homepage: www.elsevier.com/developmentalbiology Targeted disruption of the Pak5 and Pak6 genes in mice leads to deficits in learning and locomotion Tanya Nekrasova a,1, Michelle L. Jobes b,c,1, Jenhao H. Ting c, George C. Wagner c, Audrey Minden a,⁎ a Susan Lehman Cullman Laboratory for Cancer Research, Department of Chemical Biology, Ernest Mario School of Pharmacy, Rutgers, The State University of New Jersey, 164 Frelinghuysen Road, Piscataway, NJ 08854, USA b Joint Graduate Program in Toxicology, Rutgers, The State University of New Jersey, 152 Frelinghuysen Road, Piscataway, NJ 08854, USA c Department of Psychology, Rutgers, The State University of New Jersey, 152 Frelinghuysen Road, Piscataway, NJ 08854, USA article info abstract Article history: PAK6 is a member of the group B family of PAK serine/threonine kinases, and is highly expressed in the brain. Received for publication 10 September 2007 The group B PAKs, including PAK4, PAK5, and PAK6, were first identified as effector proteins for the Rho Revised 12 June 2008 GTPase Cdc42. They have important roles in filopodia formation, the extension of neurons, and cell survival. Accepted 7 July 2008 Pak4 knockout mice die in utero, and the embryos have several abnormalities, including a defect in the Available online 16 July 2008 development of motor neurons. In contrast, Pak5 knockout mice do not have any noticeable abnormalities. So far nothing is known about the biological function of Pak6. To address this, we have deleted the Pak6 gene in Keywords: Pak5 mice.
    [Show full text]
  • A Case Study Involving Hexagenia Spp
    A Molecular Evolutionary Approach for Targeted-transcriptomics of PCB Exposure: A Case Study Involving Hexagenia spp. by Gina Louise Capretta A Thesis presented to The University of Guelph In partial fulfilment of requirements for the degree of Master of Science in Integrative Biology Guelph, Ontario, Canada © Gina Capretta, DecemBer, 2015 ABSTRACT A MOLECULAR EVOLUTIONARY APPROACH FOR TARGETED- TRANSCRIPTOMICS OF PCB EXPOSURE: A CASE STUDY INVOLVING Hexagenia spp. Gina Louise Capretta Advisor: University of Guelph, 2015 Dr. Mehrdad Hajibabaei This thesis developed a methodology to target candidate xenobiotic- interacting genes conserved across taxa, as well as tested the methodology as a tool to indicate chemical exposure and elucidate possible biological effects. Using PCBs as the chemical class, this thesis identified conserved candidate PCB-interacting genes and designed and tested degenerate primers to amplify those genes in divergent species. Using next-generation sequencing technology, this thesis also investigated the targeted-transcriptome response of H. rigida, a common ecotoxicological test species, following exposure of Hexagenia spp. to PCB-52 in a 96-hour water-only test, in which survivorship and bioaccumulation were also measured. Transcript sequences of target genes were generated for H. rigida and successfully annotated. Significant down-regulation of three genes (HSP90AB1, TUBA1C, and ALDH6A1) elucidated the biological processes that may be disrupted. This research shows the potential for linking molecular events to outcomes at higher levels of biological organization, an approach relevant to environmental risk assessment. Dedication To my family iii Acknowledgements There are many people to whom I owe a debt of gratitude; their contributions helped support and encourage me throughout this journey.
    [Show full text]
  • ONCOGENOMICS Logs, Mushroom Bodies Tiny (MBT) and C45B11.1, the Importance of P21 Activated Kinase (PAK) Family Respectively (Dan Et Al., 2001)
    Oncogene (2002) 21, 3939 ± 3948 ã 2002 Nature Publishing Group All rights reserved 0950 ± 9232/02 $25.00 www.nature.com/onc Cloning and characterization of PAK5, a novel member of mammalian p21-activated kinase-II subfamily that is predominantly expressed in brain Akhilesh Pandey*,1,4, Ippeita Dan*,2,4, Troels Z Kristiansen1, Norinobu M Watanabe2, Jesper Voldby1, Eriko Kajikawa2, Roya Khosravi-Far3, Blagoy Blagoev1 and Matthias Mann*,1 1Center for Experimental Bioinformatics, University of Southern Denmark, Campusvej 55, Odense M, DK-5230, Denmark; 2Kusumi Membrane Organizer Project, ERATO, JST, Department of Biological Science, Graduate School of Science, Nagoya University, Chikusa-ku, Nagoya 464-8602 Japan; 3Department of Pathology, Beth Israel Deaconess Medical Center, Harvard Medical Center, Boston, Massachusetts, MA 02115, USA The p21-activated kinase (PAK) family of protein small GTP-binding proteins, Rac and CDC42, PAK kinases has recently attracted considerable attention as family kinases (PAKs) participate in various facets of an eector of Rho family of small G proteins and as an cellular events. In addition to modulating the organiza- upstream regulator of MAPK signalling pathways during tion of the actin cytoskeleton to control cell morphol- cellular events such as re-arrangement of the cytoskele- ogy and motility, PAKs activate mitogen-activated ton and apoptosis. We have cloned a novel human PAK protein kinase (MAPK) signalling pathways to aect family kinase that has been designated as PAK5. PAK5 gene expression (Sells and Cherno, 1997; Bagrodia contains a CDC42/Rac1 interactive binding (CRIB) and Cerione, 1999). More recently, PAKs have also motif at the N-terminus and a Ste20-like kinase domain been implicated in apoptosis and have been shown to at the C-terminus.
    [Show full text]
  • Genome-Wide Association Studies in the Japanese Population Identify Seven Novel Loci for Type 2 Diabetes
    ARTICLE Received 26 Jun 2015 | Accepted 22 Dec 2015 | Published 28 Jan 2016 DOI: 10.1038/ncomms10531 OPEN Genome-wide association studies in the Japanese population identify seven novel loci for type 2 diabetes Minako Imamura et al.# Genome-wide association studies (GWAS) have identified more than 80 susceptibility loci for type 2 diabetes (T2D), but most of its heritability still remains to be elucidated. In this study, we conducted a meta-analysis of GWAS for T2D in the Japanese population. Combined data from discovery and subsequent validation analyses (23,399 T2D cases and 31,722 controls) identify 7 new loci with genome-wide significance (Po5 Â 10 À 8), rs1116357 near CCDC85A, rs147538848 in FAM60A, rs1575972 near DMRTA1, rs9309245 near ASB3, rs67156297 near ATP8B2, rs7107784 near MIR4686 and rs67839313 near INAFM2. Of these, the association of 4 loci with T2D is replicated in multi-ethnic populations other than Japanese (up to 65,936 T2Ds and 158,030 controls, Po0.007). These results indicate that expansion of single ethnic GWAS is still useful to identify novel susceptibility loci to complex traits not only for ethnicity-specific loci but also for common loci across different ethnicities. Correspondence and requests for materials should be addressed to S.M. (email: [email protected]) or to T.K. (email: [email protected]). #A full list of authors and their affiliations appears at the end of the paper. NATURE COMMUNICATIONS | 7:10531 | DOI: 10.1038/ncomms10531 | www.nature.com/naturecommunications 1 ARTICLE NATURE COMMUNICATIONS | DOI: 10.1038/ncomms10531 o date, more than 80 susceptibility loci for type 2 diabetes association with T2D (Po1 Â 10 À 6).
    [Show full text]
  • Integrated Post-GWAS Analysis Shed New Light on the Disease
    Genetics: Early Online, published on October 17, 2016 as 10.1534/genetics.116.187195 Integrated post-GWAS analysis shed new light on the disease mechanisms of schizophrenia Jhih-Rong Lin1, Ying Cai1, Quanwei Zhang1, Wen Zhang1, Rubén Nogales-Cadenas1, Zhengdong D. Zhang1,§ 1Department of Genetics, Albert Einstein College of Medicine, Bronx, NY 10461, USA §Corresponding author (E-mail: [email protected]) Keywords: schizophrenia; GWAS; disease risk gene prioritization 1 Copyright 2016. ABSTRACT Schizophrenia is a severe mental disorder with a large genetic component. Recent genome-wide association studies (GWAS) have identified many schizophrenia-associated common variants. For most of the reported associations, however, the underlying biological mechanisms are not clear. The critical first step for their elucidation is to identify the most likely disease genes as the source of the association signals. Here, we describe a general computational framework of post- GWAS analysis for complex disease gene prioritization. We identify 132 putative schizophrenia risk genes in 76 risk regions spanning 120 schizophrenia-associated common variants, 78 of which have not been recognized as schizophrenia disease genes by previous GWAS. Even more significantly, 29 of them are outside the risk regions, likely under regulation of transcriptional regulatory elements therein contained. These putative schizophrenia risk genes are transcriptionally active in both brain and the immune system and highly enriched among cellular pathways, consistent with leading pathophysiological hypotheses about the pathogenesis of schizophrenia. With their involvement in distinct biological processes, these putative schizophrenia risk genes with different association strengths show distinctive temporal expression patterns and play specific biological roles during brain development.
    [Show full text]
  • 1 Supplemental Table 1. Comparison Among Q-RT-PCR, ISH-TMA And
    Supplemental Table 1. Comparison among Q-RT-PCR, ISH-TMA and IHC for the detection of ErbB-2 in breast cancers. IHC ISH-TMA Q-RT-PCR Q-RT-PCR range Q-RT-PCR score 0 0 0* 0 0 0 0* 0 0 0 1 0 0 0 1 0 0 0 1 0 0 1 2 0 1 1 2 0 0 0 3 0 0 0 3 0 0 0 3 0 0 0 3 0 0 0 3 x<10 0 1 1 3 0 0 0 4 0 0 0 5 0 0 0 5 0 0 0 5 0 1 1 5 0 0 0 6 0 0 0 8 0 0 0 8 0 1 1 8 0 0 1 8 0 1 1 11 1 1 1 11 1 1 1 11 1 1 0 12 1 1 1 13 10<x<20 1 0 1 13 1 1 1 16 1 1 1 17 1 2 1 17 1 2 2 22 2 2 2 43 2 20<x<100 3 2 74 2 3 2 86 2 3 3 213 3 3 3 259 x>100 3 3 3 362 3 Legend to Supplemental Table 1. This experiment was set up to demonstrate that there is good semi-quantitative correlation between the levels of expression detected by ISH-TMA, IHC and Q-RT-PCR. We compared the three methods on levels of ErbB-2 expression in breast cancer, since ErbB-2 is overexpressed in breast cancers, over a wide range of levels.
    [Show full text]
  • Unraveling the Molecular Mechanism Underlying ALS-Linked Astrocyte
    Unraveling the molecular mechanism underlying ALS- linked astrocyte toxicity for motor neurons Burcin Ikiz Submitted in partial fulfillment of the requirements for the degree of Doctor of Philosophy under the Executive Committee of the Graduate School of Arts and Sciences COLUMBIA UNIVERSITY 2013 © 2013 Burcin Ikiz All rights reserved Abstract Unraveling the molecular mechanism underlying ALS-linked astrocyte toxicity for motor neurons Burcin Ikiz Mutations in superoxide dismutase-1 (SOD1) cause a familial form of amyotrophic lateral sclerosis (ALS), a fatal paralytic disorder. Transgenic mutant SOD1 rodents capture the hallmarks of this disease, which is characterized by a progressive loss of motor neurons. Studies in chimeric and conditional transgenic mutant SOD1 mice indicate that non-neuronal cells, such as astrocytes, play an important role in motor neuron degeneration. Consistent with this non-cell autonomous scenario are the demonstrations that wild-type primary and embryonic stem cell- derived motor neurons selectively degenerate when cultured in the presence of either mutant SOD1-expressing astrocytes or medium conditioned with such mutant astrocytes. The work in this thesis rests on the use of an unbiased genomic strategy that combines RNA-Seq and “reverse gene engineering” algorithms in an attempt to decipher the molecular underpinnings of motor neuron degeneration caused by mutant astrocytes. To allow such analyses, first, mutant SOD1- induced toxicity on purified embryonic stem cell-derived motor neurons was validated and characterized. This was followed by the validation of signaling pathways identified by bioinformatics in purified embryonic stem cell-derived motor neurons, using both pharmacological and genetic techniques, leading to the discovery that nuclear factor kappa B (NF-κB) is instrumental in the demise of motor neurons exposed to mutant astrocytes in vitro.
    [Show full text]
  • Transcriptional Maturation of the Mouse Auditory Forebrain Troy A
    Hackett et al. BMC Genomics (2015) 16:606 DOI 10.1186/s12864-015-1709-8 RESEARCH ARTICLE Open Access Transcriptional maturation of the mouse auditory forebrain Troy A. Hackett1,8*, Yan Guo4, Amanda Clause2, Nicholas J. Hackett3, Krassimira Garbett5, Pan Zhang4, Daniel B. Polley2 and Karoly Mirnics5,6,7,8 Abstract Background: The maturation of the brain involves the coordinated expression of thousands of genes, proteins and regulatory elements over time. In sensory pathways, gene expression profiles are modified by age and sensory experience in a manner that differs between brain regions and cell types. In the auditory system of altricial animals, neuronal activity increases markedly after the opening of the ear canals, initiating events that culminate in the maturation of auditory circuitry in the brain. This window provides a unique opportunity to study how gene expression patterns are modified by the onset of sensory experience through maturity. As a tool for capturing these features, next-generation sequencing of total RNA (RNAseq) has tremendous utility, because the entire transcriptome can be screened to index expression of any gene. To date, whole transcriptome profiles have not been generated for any central auditory structure in any species at any age. In the present study, RNAseq was used to profile two regions of the mouse auditory forebrain (A1, primary auditory cortex; MG, medial geniculate) at key stages of postnatal development (P7, P14, P21, adult) before and after the onset of hearing (~P12). Hierarchical clustering, differential expression, and functional geneset enrichment analyses (GSEA) were used to profile the expression patterns of all genes. Selected genesets related to neurotransmission, developmental plasticity, critical periods and brain structure were highlighted.
    [Show full text]
  • P21-ACTIVATED KINASE: a NOVEL THERAPUETIC TARGET of CELECOXIB and THYROID CANCER
    p21-ACTIVATED KINASE: A NOVEL THERAPUETIC TARGET IN THYROID CANCER DISSERTATION Presented in Partial Fulfillment of the Requirements for the Degree Doctor of Philosophy in the Graduate School of The Ohio State University By Leonardo M. Porchia, M.S. * * * * * The Ohio State University 2007 Dissertation Committee: Professor Matthew D. Ringel, Advisor Approved by Professor Robert Brueggemeier Professor Ching-Shih Chen Professor Lawrence S. Kirschner Advisor Ohio State Biochemistry Program ABSTRACT Follicular cell derived thyroid cancer (i.e. follicular, papillary and anaplastic thyroid cancer) is the most common endocrine malignancy. While patients with diagnosed with early stage disease have an excellent prognosis, patients with invasive or metastatic thyroid cancer have poor survival rates. Because progressive thyroid cancer is unresponsive to chemotherapy, there is a critical need to identify novel therapeutic targets. Genetic alterations that result in enhanced activation of the RAS-RAF-MEK and PI3K-AKT pathways occur in more than 50% of papillary (PTC), follicular (FTC), and anaplastic (ATC) thyroid cancers. However, the key regulators of thyroid cancer invasion and metastases are less certain. Many lines of evidence suggest important roles for PI3K signaling and the process of epithelial-to-mesenchymal transition (EMT) in thyroid cancer progression. Thus, we are working to develop inhibitors of these pathways for thyroid cancer. OSU-03012 (OSU) is a celecoxib derivative that was optimized to inhibit PDK-1, a key signaling kinase in the PI3K cascade. NPA (papillary), WRO (follicular), and ARO (anaplastic) thyroid cancer cell lines were used to study the effects of OSU on thyroid cancer cells in vitro. OSU inhibited proliferation and induced cytotoxicity at doses sufficient to inhibit PDK-1- mediated AKT phosphorylation.
    [Show full text]
  • UNIVERSITY of CALIFORNIA, SAN DIEGO Accurate Prediction Of
    UNIVERSITY OF CALIFORNIA, SAN DIEGO Accurate Prediction of Causative Protein Kinase Polymorphisms in Inherited Disease and Cancer A Dissertation submitted in partial satisfaction of the Requirements for the degree Doctor of Philosophy in Biomedical Sciences by Ali Torkamani Committee in charge: Professor Nicholas Schork, Chair Professor Arshad Desai Professor Gerard Manning Professor Alexandra Newton Professor Susan Taylor Professor Anthony Wynshaw-Boris 2008 Copyright Ali Torkamani, 2008 All rights reserved. The Dissertation of Ali Torkamani is approved and it is acceptable in quality and form for publication on microfilm: Chair University of California, San Diego 2008 iii DEDICATION I dedicate this dissertation to my loving mother, Mitra Moassessi, and father, Naser Torkamani, whose love, support, and encouragement made this work possible. iv EPIGRAPH Dreaming when Dawn's Left Hand was in the Sky I heard a Voice within the Tavern cry, "Awake, my Little ones, and fill the Cup Before Life's Liquor in its Cup be dry." Omar Khayyam v TABLE OF CONTENTS Signature Page………………………………………………………………... iii Dedication…………………………………………………………………...... iv Epigraph……………………………………………………………………..... v Table of Contents……………………………………………………………... vi List of Abbreviations………………………………………………………..... ix List of Figures………………………………………………………………… x List of Tables………………………………………………………………..... xiii Acknowledgements…………………………………………………………… xvi Vita………………………………………………………………………….... xviii Abstract……………………………………………………………………….. xix Introduction…………………………………………………………………...
    [Show full text]