DOCSLIB.ORG

P21-ACTIVATED KINASE: a NOVEL THERAPUETIC TARGET of CELECOXIB and THYROID CANCER

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read 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. Unexpectedly, OSU inhibited cell motility in NPA cells at doses below its IC50 for PDK-1 and below those ii sufficient to reduce AKT phosphorylation, suggesting that that the inhibition of migration by OSU might be due to another mechanism. p21 activated kinases (PAK) are master regulators of cell motility and EMT that are regulated both by PDK1 and other signaling cascades. Phospho-PAK levels were reduced with 1 μM of OSU in motile NPA cells and WRO cells. PAK-dependent phosphorylation of vimentin, a key regulator of thyroid cell EMT, was decreased at similar doses, consistent with reduced activity in these cells. Subsequent in vitro kinase assays demonstrated that OSU competitively inhibits ATP binding to PAK. Finally, overexpression of constitutively active-PAK1 rescued the anti- migratory effects of OSU. These data demonstrate that PAK is a novel target of and that OSU might be a therapeutic option for tumors with PAK-dependent invasion and motility. Because of these findings, and because prior work by our group suggested that signaling through PAK might be dysregulated in thyroid cancer invasion, we undertook a study to determine if PAK expression or PAK activity were altered in thyroid cancer. Protein and total RNA was isolated from ten PTC tissue samples and from the normal tissue in the opposite lobes for Western blot and quantitative RT-PCR of all PAK isoforms. Of the six isoforms, PAK2 and PAK4 mRNA and protein expression levels were increased in the majority of PTCs. In addition, the levels of phosphorylated PAK were higher in 8 of 10 PTCs. Finally, molecular inhibition of PAK using cDNAs designed to inhibition PAKs1, 2, and 3 confirmed that thyroid cancer cells display PAK- dependent cell migration. Taken together, these data support a role for PAK activity in thyroid cancer and suggest it might be an appropriate therapeutic target for this disease in patients. iii The most common mutation of PTC is BRAF V600E. Expression of this mutant BRAF induces thyroid cell invasion and is associated with a more aggressive form of PTC clinically. PAK is known to functionally phosphorylate C-RAF, a closely related RAF family protein member, at serine 338. Nine potential PAK phosphorylation sites were found in the B-RAF protein sequence. One site aligned to serine 338 in C-RAF, suggesting this site might represent a PAK phosphorylation site on BRAF. Subsequent experiments demonstrated that B-RAF and PAK co-immunoprecipitate, suggesting they interact directly. This interaction was identified both in overexpression and endogenous systems. Further studies are planned to clarify if the BRAF-PAK interactions are functional and to identify specific phosphorylation sites Histone deacetylase inhibitors (HDACi) are a class of agents with broad cellular effects. HDAC42 is a novel HDACi that has been shown to be cytotoxic in a number of cell systems. This agent has also been shown to regulate expression and function of signaling molecules, including AKT. HDAC42 inhibited thyroid cancer cell line proliferation, and was cytotoxic at 48 hours. Distinct from OSU, this compound did not appear to inhibit migration. Treatment with HDAC42 increased acetyl-histone 4 at 24 hours and the level of AKT expression and activation were inhibited. However, while levels of PAK protein were reduced after 48 hours of exposure; levels of phospho-PAK at 24 hours were unaltered and increased at 48 hours. Based on these results, we hypothesize that the lack of PAK inhibition may be responsible for the inability of HDAC42 to block migration, suggesting that the combination of PAK and HDAC inhibitors could be a novel treatment for metastatic cancer. iv In summary, these results are the first to demonstrate that PAK is a novel target of OSU, and its inhibition is at least partially responsible for in vitro effects on migration. They are also the first to determine that PTCs are characterized by overexpression of PAKs 2 and -4, which PAK activity is frequently enhanced in PTC and is functionally involved in PTC cell biology in vitro. The additional finding that BRAF and PAK physically interact will lead to new experiments determining if there is an important role for interactions between these signaling molecules in PTC biology. Finally, the finding that HDAC42 increases phospho-PAK levels suggest that combination therapy with this OSU and this compound may be a rationale approach to thyroid cancer therapy. These results suggest that PAK may represent a novel therapeutic target for thyroid cancer, and that compounds based on OSU may be optimized to inhibit this pathway. v Dedicated to my mother and father and family vi ACKNOWLEDGMENTS • I would like to acknowledge Dr. Ringel for his guidance, immense understanding and support, and providing me the ability to work in his lab. • I would also like to acknowledge Dr. Chen for his guidance and support and originally giving the opportunity to study signal transduction and get a better prospective in a drug discovery lab. • Dr. Jiuxiang Zhu, whose help with planning experiments and analyzing data and leading me in the beginning. • Colleagues and friends: Dr. Sam Kulp, Dr. Motoyasu Saji, Dr. Allan Espinosa, Marcy Geurra, Dr. Motoo Shinohara, Dr. Jun Yea Chung, Yu-Chieh Wang, Ho- Pi Lin, Ya-ting Wang, for your help with experiments, analyzing data, and making lab an enjoyable experience. • Yunlong Zhang for his help with the computer modeling. • Susanna Pearce for helping edit and revising my writings. vii VITA 1998 – 2002 B.S Biochemistry Otterbein College, Westerville Ohio 2002 – 2006 M.S. Biochemistry The Ohio State University, Columbus, Ohio 2002 – Present Graduate Teaching and Research Associate College of Chemistry, The Ohio State University College of Pharmacy, The Ohio State University College of Medicine, The Ohio State University PUBLICATIONS 1. Porchia LM, Guerra M, Wang YC, Zhang Y, Espinosa AV, Shinohara M, Kulp SK, Kirschner LS, Saji M, Chen CS, Ringel MD. “OSU03012, A Celecoxib Derivative, Directly Targets p21 Activated Kinase.” Mol Pharmacol. 2007 Aug 2; [Epub] 2. Espinosa AV, Porchia L and Ringel MD. “Targeting BRAF in thyroid cancer.” British Journal of Cancer. 2007 Jan 15:96(1): 16-20. FIELDS OF STUDY Major Field: Biochemistry viii TABLE OF CONTENTS Page Abstract................................................................................................................................ii Dedication...........................................................................................................................iii Acknowledgments.............................................................................................................vii Vita...................................................................................................................................viii List of Tables.....................................................................................................................xii List of Figure....................................................................................................................xiii Abbreviations.....................................................................................................................xv Chapter 1 – Introduction......................................................................................................1 1.1 Normal Thyroid Function..................................................................................2 1.2 Thyroid Cancer .................................................................................................3 1.2.1 Oncogenic Causes of Thyroid Cancer................................................5 1.2.1.1 RET/PTC and B-RAF Alteration Lead to the Development
Load more

Recommended publications
  • A Computational Approach for Defining a Signature of Β-Cell Golgi Stress in Diabetes Mellitus
    Page 1 of 781 Diabetes A Computational Approach for Defining a Signature of β-Cell Golgi Stress in Diabetes Mellitus Robert N. Bone1,6,7, Olufunmilola Oyebamiji2, Sayali Talware2, Sharmila Selvaraj2, Preethi Krishnan3,6, Farooq Syed1,6,7, Huanmei Wu2, Carmella Evans-Molina 1,3,4,5,6,7,8* Departments of 1Pediatrics, 3Medicine, 4Anatomy, Cell Biology & Physiology, 5Biochemistry & Molecular Biology, the 6Center for Diabetes & Metabolic Diseases, and the 7Herman B. Wells Center for Pediatric Research, Indiana University School of Medicine, Indianapolis, IN 46202; 2Department of BioHealth Informatics, Indiana University-Purdue University Indianapolis, Indianapolis, IN, 46202; 8Roudebush VA Medical Center, Indianapolis, IN 46202. *Corresponding Author(s): Carmella Evans-Molina, MD, PhD ([email protected]) Indiana University School of Medicine, 635 Barnhill Drive, MS 2031A, Indianapolis, IN 46202, Telephone: (317) 274-4145, Fax (317) 274-4107 Running Title: Golgi Stress Response in Diabetes Word Count: 4358 Number of Figures: 6 Keywords: Golgi apparatus stress, Islets, β cell, Type 1 diabetes, Type 2 diabetes 1 Diabetes Publish Ahead of Print, published online August 20, 2020 Diabetes Page 2 of 781 ABSTRACT The Golgi apparatus (GA) is an important site of insulin processing and granule maturation, but whether GA organelle dysfunction and GA stress are present in the diabetic β-cell has not been tested. We utilized an informatics-based approach to develop a transcriptional signature of β-cell GA stress using existing RNA sequencing and microarray datasets generated using human islets from donors with diabetes and islets where type 1(T1D) and type 2 diabetes (T2D) had been modeled ex vivo. To narrow our results to GA-specific genes, we applied a filter set of 1,030 genes accepted as GA associated.
    [Show full text]
  • 4-6 Weeks Old Female C57BL/6 Mice Obtained from Jackson Labs Were Used for Cell Isolation
    Methods Mice: 4-6 weeks old female C57BL/6 mice obtained from Jackson labs were used for cell isolation. Female Foxp3-IRES-GFP reporter mice (1), backcrossed to B6/C57 background for 10 generations, were used for the isolation of naïve CD4 and naïve CD8 cells for the RNAseq experiments. The mice were housed in pathogen-free animal facility in the La Jolla Institute for Allergy and Immunology and were used according to protocols approved by the Institutional Animal Care and use Committee. Preparation of cells: Subsets of thymocytes were isolated by cell sorting as previously described (2), after cell surface staining using CD4 (GK1.5), CD8 (53-6.7), CD3ε (145- 2C11), CD24 (M1/69) (all from Biolegend). DP cells: CD4+CD8 int/hi; CD4 SP cells: CD4CD3 hi, CD24 int/lo; CD8 SP cells: CD8 int/hi CD4 CD3 hi, CD24 int/lo (Fig S2). Peripheral subsets were isolated after pooling spleen and lymph nodes. T cells were enriched by negative isolation using Dynabeads (Dynabeads untouched mouse T cells, 11413D, Invitrogen). After surface staining for CD4 (GK1.5), CD8 (53-6.7), CD62L (MEL-14), CD25 (PC61) and CD44 (IM7), naïve CD4+CD62L hiCD25-CD44lo and naïve CD8+CD62L hiCD25-CD44lo were obtained by sorting (BD FACS Aria). Additionally, for the RNAseq experiments, CD4 and CD8 naïve cells were isolated by sorting T cells from the Foxp3- IRES-GFP mice: CD4+CD62LhiCD25–CD44lo GFP(FOXP3)– and CD8+CD62LhiCD25– CD44lo GFP(FOXP3)– (antibodies were from Biolegend). In some cases, naïve CD4 cells were cultured in vitro under Th1 or Th2 polarizing conditions (3, 4).
    [Show full text]
  • 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]
  • Effect of STAT3 Inhibition on the Metabolic Switch in a Highly STAT3-Activated Lymphoma Cell Line
    CANCER GENOMICS & PROTEOMICS 12 : 133-142 (2015) Effect of STAT3 Inhibition on the Metabolic Switch in a Highly STAT3-activated Lymphoma Cell Line YASUTO AKIYAMA 1* , AKIRA IIZUKA 1* , AKIKO KUME 1, MASARU KOMIYAMA 1, KENICHI URAKAMI 2, TADASHI ASHIZAWA 1, HARUO MIYATA 1, MAHO OMIYA 1, MASATOSHI KUSUHARA 3 and KEN YAMAGUCHI 4 1Immunotherapy Division, 2Cancer Diagnostics Division, 3Regional Resources Division, Shizuoka Cancer Center Research Institute, Sunto-gun, Shizuoka, Japan; 4Office of the President, Shizuoka Cancer Center Hospital, Sunto-gun, Shizuoka, Japan Abstract. Background: Signal transducer and activator of enzymes including fructose-bisphosphate aldolase A transcription (STAT)3 is involved in a metabolic shift in (ALDOA) as a metabolic marker candidate for STAT3- cancer cells, the Warburg effect through its pro-oncogenic targeting therapy using STAT3-specific shRNA gene activity. To develop efficient STAT3 inhibitors against cancer transduction. In particular, latexin expression was up- cells, novel proteomic and metabolic target molecules need regulated in four STAT3-activated cancer cell lines including to be explored using multi-omics approaches in the context of SCC-3 transduced with STAT3-specific shRNA. The up- STAT3 gene inhibition-mediated tumor growth suppression. regulation of latexin was identified in SCC-3 tumors Materials and Methods: We found that short hairpin transplanted to nude mice after treatment with STAT3 (sh)RNA-mediated STAT3 inhibition suppressed tumor inhibitor. Conclusion: Our results suggest that STAT3 growth in a highly STAT3-activated lymphoma cell line, inactivation reverses the glycolytic shift by down-regulating SCC-3 cells, and we investigated the effect of STAT3 key enzymes and that it induces up-regulation of latexin as a inhibition on metabolic switching using 2-dimensional tumor-suppressor molecule, which partially results in cancer differential gel electrophoresis and capillary electrophoresis- cell apoptosis and tumor growth suppression.
    [Show full text]
  • Ubiquitin and Ubiquitin-Like Proteins Are Essential Regulators of DNA Damage Bypass
    cancers Review Ubiquitin and Ubiquitin-Like Proteins Are Essential Regulators of DNA Damage Bypass Nicole A. Wilkinson y, Katherine S. Mnuskin y, Nicholas W. Ashton * and Roger Woodgate * Laboratory of Genomic Integrity, National Institute of Child Health and Human Development, National Institutes of Health, 9800 Medical Center Drive, Rockville, MD 20850, USA; [email protected] (N.A.W.); [email protected] (K.S.M.) * Correspondence: [email protected] (N.W.A.); [email protected] (R.W.); Tel.: +1-301-435-1115 (N.W.A.); +1-301-435-0740 (R.W.) Co-first authors. y Received: 29 August 2020; Accepted: 29 September 2020; Published: 2 October 2020 Simple Summary: Ubiquitin and ubiquitin-like proteins are conjugated to many other proteins within the cell, to regulate their stability, localization, and activity. These modifications are essential for normal cellular function and the disruption of these processes contributes to numerous cancer types. In this review, we discuss how ubiquitin and ubiquitin-like proteins regulate the specialized replication pathways of DNA damage bypass, as well as how the disruption of these processes can contribute to cancer development. We also discuss how cancer cell survival relies on DNA damage bypass, and how targeting the regulation of these pathways by ubiquitin and ubiquitin-like proteins might be an effective strategy in anti-cancer therapies. Abstract: Many endogenous and exogenous factors can induce genomic instability in human cells, in the form of DNA damage and mutations, that predispose them to cancer development. Normal cells rely on DNA damage bypass pathways such as translesion synthesis (TLS) and template switching (TS) to replicate past lesions that might otherwise result in prolonged replication stress and lethal double-strand breaks (DSBs).
    [Show full text]
  • (12) Patent Application Publication (10) Pub. No.: US 2003/0082511 A1 Brown Et Al
    US 20030082511A1 (19) United States (12) Patent Application Publication (10) Pub. No.: US 2003/0082511 A1 Brown et al. (43) Pub. Date: May 1, 2003 (54) IDENTIFICATION OF MODULATORY Publication Classification MOLECULES USING INDUCIBLE PROMOTERS (51) Int. Cl." ............................... C12O 1/00; C12O 1/68 (52) U.S. Cl. ..................................................... 435/4; 435/6 (76) Inventors: Steven J. Brown, San Diego, CA (US); Damien J. Dunnington, San Diego, CA (US); Imran Clark, San Diego, CA (57) ABSTRACT (US) Correspondence Address: Methods for identifying an ion channel modulator, a target David B. Waller & Associates membrane receptor modulator molecule, and other modula 5677 Oberlin Drive tory molecules are disclosed, as well as cells and vectors for Suit 214 use in those methods. A polynucleotide encoding target is San Diego, CA 92121 (US) provided in a cell under control of an inducible promoter, and candidate modulatory molecules are contacted with the (21) Appl. No.: 09/965,201 cell after induction of the promoter to ascertain whether a change in a measurable physiological parameter occurs as a (22) Filed: Sep. 25, 2001 result of the candidate modulatory molecule. Patent Application Publication May 1, 2003 Sheet 1 of 8 US 2003/0082511 A1 KCNC1 cDNA F.G. 1 Patent Application Publication May 1, 2003 Sheet 2 of 8 US 2003/0082511 A1 49 - -9 G C EH H EH N t R M h so as se W M M MP N FIG.2 Patent Application Publication May 1, 2003 Sheet 3 of 8 US 2003/0082511 A1 FG. 3 Patent Application Publication May 1, 2003 Sheet 4 of 8 US 2003/0082511 A1 KCNC1 ITREXCHO KC 150 mM KC 2000000 so 100 mM induced Uninduced Steady state O 100 200 300 400 500 600 700 Time (seconds) FIG.
    [Show full text]
  • Involvement of Inhibitor Kappa B Kinase 2 (IKK2) in the Regulation of Vascular Tone
    Laboratory Investigation (2018) 98:1311–1319 https://doi.org/10.1038/s41374-018-0061-4 ARTICLE Involvement of inhibitor kappa B kinase 2 (IKK2) in the regulation of vascular tone 1 1 1 1 Youngin Kwon ● Soo-Kyoung Choi ● Seonhee Byeon ● Young-Ho Lee Received: 6 November 2017 / Revised: 22 March 2018 / Accepted: 23 March 2018 / Published online: 21 May 2018 © United States & Canadian Academy of Pathology 2018 Abstract Inhibitor kappa B kinase 2 (IKK2) plays an essential role in the activation of nuclear factor kappa B (NF-kB). Recently, it has been suggested that IKK2 acts as a myosin light chain kinase (MLCK) and contributes to vasoconstriction in mouse aorta. However, the underlying mechanisms are still unknown. Therefore, we investigated whether IKK2 acts as a MLCK or regulates the activity of myosin light chain phosphatase (MLCP). Pressure myograph was used to measure vascular tone in rat mesenteric arteries. Immunofluorescence staining was performed to identify phosphorylation levels of MLC (ser19), MYPT1 (thr853 and thr696) and CPI-17 (thr38). SC-514 (IKK2 inhibitor, 50 μM) induced relaxation in the mesenteric arteries pre-contracted with 70 mM high K+ solution or U-46619 (thromboxane analog, 5 μM). The relaxation induced by SC-514 + 1234567890();,: 1234567890();,: was increased in the arteries pre-contracted with U-46619 compared to arteries pre-contracted with 70 mM high K solution. U-46619-induced contraction was decreased by treatment of SC-514 in the presence of MLCK inhibitor, ML-7 (10 μM). In the absence of intracellular Ca2+, U-46619 still induced contraction, which was decreased by treatment of SC-514.
    [Show 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.
    [Show full text]
  • STAT3-Mediated Metabolic Reprograming in Cellular Transformation and Implications for Drug Resistance
    MINI REVIEW published: 08 June 2015 doi: 10.3389/fonc.2015.00121 STAT3-mediated metabolic reprograming in cellular transformation and implications for drug resistance Valeria Poli* and Annalisa Camporeale* Department of Molecular Biotechnology and Health Sciences, Molecular Biotechnology Center, University of Torino, Torino, Italy Signal transducer and activator of transcription (STAT)3 mediates the signaling down- stream of cytokine and growth factor receptors, regulating the expression of target genes. It is constitutively phosphorylated on tyrosine (Y-P) in many tumors, where its transcrip- tional activity can induce a metabolic switch toward aerobic glycolysis and down-regulate mitochondrial activity, a prominent metabolic feature of most cancer cells, correlating Edited by: Gavin P. McStay, with reduced production of ROS, delayed senescence, and protection from apoptosis. New York Institute of Technology, USA STAT3 can, however, also localize to mitochondria, where its serine-phosphorylated Reviewed by: (S-P) form preserves mitochondrial oxidative phosphorylation and controls the opening Masaaki Murakami, of the mitochondrial permeability transition pore, also promoting survival and resistance Hokkaido University, Japan Yun Dai, to apoptosis in response to specific signals/oncogenes such as RAS. Thus, downstream Virginia Commonwealth University of different signals, both nuclear, Y-P STAT3, and mitochondrial, S-P STAT3, can act by and Massey Cancer Center, USA promoting cell survival and reducing ROS production. Here, we discuss
    [Show full text]
  • UC San Diego UC San Diego Electronic Theses and Dissertations
    UC San Diego UC San Diego Electronic Theses and Dissertations Title Insights from reconstructing cellular networks in transcription, stress, and cancer Permalink https://escholarship.org/uc/item/6s97497m Authors Ke, Eugene Yunghung Ke, Eugene Yunghung Publication Date 2012 Peer reviewed|Thesis/dissertation eScholarship.org Powered by the California Digital Library University of California UNIVERSITY OF CALIFORNIA, SAN DIEGO Insights from Reconstructing Cellular Networks in Transcription, Stress, and Cancer A dissertation submitted in the partial satisfaction of the requirements for the degree Doctor of Philosophy in Bioinformatics and Systems Biology by Eugene Yunghung Ke Committee in charge: Professor Shankar Subramaniam, Chair Professor Inder Verma, Co-Chair Professor Web Cavenee Professor Alexander Hoffmann Professor Bing Ren 2012 The Dissertation of Eugene Yunghung Ke is approved, and it is acceptable in quality and form for the publication on microfilm and electronically ________________________________________________________________ ________________________________________________________________ ________________________________________________________________ ________________________________________________________________ Co-Chair ________________________________________________________________ Chair University of California, San Diego 2012 iii DEDICATION To my parents, Victor and Tai-Lee Ke iv EPIGRAPH [T]here are known knowns; there are things we know we know. We also know there are known unknowns; that is to say we know there
    [Show full text]
  • Microrna Regulation and Human Protein Kinase Genes
    MICRORNA REGULATION AND HUMAN PROTEIN KINASE GENES REQUIRED FOR INFLUENZA VIRUS REPLICATION by LAUREN ELIZABETH ANDERSEN (Under the Direction of Ralph A. Tripp) ABSTRACT Human protein kinases (HPKs) have profound effects on cellular responses. To better understand the role of HPKs and the signaling networks that influence influenza replication, a siRNA screen of 720 HPKs was performed. From the screen, 17 “hit” HPKs (NPR2, MAP3K1, DYRK3, EPHA6, TPK1, PDK2, EXOSC10, NEK8, PLK4, SGK3, NEK3, PANK4, ITPKB, CDC2L5, CALM2, PKN3, and HK2) were validated as important for A/WSN/33 influenza virus replication, and 6 HPKs (CDC2L5, HK2, NEK3, PANK4, PLK4 and SGK3) identified as important for A/New Caledonia/20/99 influenza virus replication. Meta-analysis of the hit HPK genes identified important for influenza virus replication showed a level of overlap, most notably with the p53/DNA damage pathway. In addition, microRNAs (miRNAs) predicted to target the validated HPK genes were determined based on miRNA seed site predictions from computational analysis and then validated using a panel of miRNA agonists and antagonists. The results identify miRNA regulation of hit HPK genes identified, specifically miR-148a by targeting CDC2L5 and miR-181b by targeting SGK3, and suggest these miRNAs also have a role in regulating influenza virus replication. Together these data advance our understanding of miRNA regulation of genes critical for virus replication and are important for development novel influenza intervention strategies. INDEX WORDS: Influenza virus,
    [Show full text]
  • BIOGRAPHICAL SKETCH NAME: Mitchell, Beverly
    OMB No. 0925-0001/0002 (Rev. 08/12 Approved Through 8/31/2015) BIOGRAPHICAL SKETCH Provide the following information for the Senior/key personnel and other significant contributors. Follow this format for each person. DO NOT EXCEED FIVE PAGES. NAME: Mitchell, Beverly S. eRA COMMONS USER NAME (credential, e.g., agency login): BMITCHELL POSITION TITLE: George E. Becker Professor of Medicine Director, Stanford Cancer Institute EDUCATION/TRAINING DEGREE Completion (if FIELD OF STUDY INSTITUTION AND LOCATION Date applicable) Smith College, Northampton, MA A.B. 1965 Biochemistry Harvard Medical School, Boston, MA M.D. 1969 Medicine University of Washington, Seattle, WA Resident 1969-72 Internal Medicine University of Zurich, Zurich, Switzerland Fellow 1973-75 Metabolism University of Michigan, Ann Arbor, MI Fellow 1975-77 Hematology/Oncology A. Personal Statement I have had a career-long research interest in the role of purine metabolic pathways in the regulation of cell growth and differentiation, with particular emphasis on applications to the treatment of acute leukemias. More recently, we have focused on the role of specific pathogenic mutations, such as those in the C- terminus of nucleophosmin, in generating responses to agents that induce reactive oxygen species, as well as on the epigenetic events and signaling pathways that regulate ribosomal RNA synthesis in leukemic cells. I have also led a LLS SCOR focused on finding new treatments for MDS. With this background, I am delighted to service as an internal advisor for the NCI Leukemia
    [Show full text]