DOCSLIB.ORG

The Identification of Tumour Antigens Recognized by Patients with Duke's B

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
The Identification of Tumour Antigens Recognized by Patients with Duke's B Title: The identification of tumour antigens recognized by patients with Duke’s B (Stage II) reactive colorectal cancers using SEREX Name: Viktoriya Bogdanova Boncheva This is a digitised version of a dissertation submitted to the University of Bedfordshire. It is available to view only. This item is subject to copyright. The identification of tumour antigens recognized by patients with Duke’s B (Stage II) reactive colorectal cancers using SEREX by Viktoriya Bogdanova Boncheva A thesis submitted to the Faculty of Creative Arts, Technologies & Science, University of Bedfordshire, in fulfilment of the requirements for the degree of Master of Philosophy. Submitted December 2013 ABSTRACT Colorectal cancer (CRC) is one of the most commonly diagnosed cancers in both men and women, posing a serious demographic and economic burden worldwide. In the UK, CRC affects one in every twenty people and it is often detected once well-established and after it has spread beyond the bowel (Stage IIA-C and Stage IIIA-C). A diagnosis at such advanced stages is associated with poor treatment response and survival. However, studies have identified two sub-groups of post-treatment CRC patients – those with good outcome (reactive disease) and those with poor outcome (non- reactive disease). Evidence indicates the presence of an effective immune response differentiates between those patients who respond well to treatment and those who do not. To investigate these underlying mechanisms we used the serological analysis of cDNA recombinant libraries (SEREX) technique to determine which antigens are recognised by patients in each group. Immunoscreening a healthy donor testes cDNA library with sera from three patients with Duke’s B reactive disease led to the identification of five antigens. These were (1) the immunoglobulin heavy constant gamma 3 (G3m marker), IGHG3 gene, located on chromosome 14 at 14q32.33, which encodes IgG3, and was recognised by sera from patients CC005 and CC014; (2) the immunoglobulin heavy constant gamma 2, IGHG2 gene, located on chromosome 14 at 14q32.33 and recognised by CC014 sera; while (3) CYB5R3, (4) RPL37A and (5) SLC34A2 were recognised by CC005, CC014 and CC014 sera respectively. CYB5R3 is a NADH-cytochrome b5 reductase 3 protein which has been shown to be upregulated in lung tissue with a RAS mutation in mice. Ribosomal protein L37a (RPL37A) has previously been shown to be upregulated in astrocytomas and to have a general association with lifetime glioblastoma survival and overall glioblastoma survival. Solute carrier family 34 member 2 (SLC34A2) encodes a protein which acts as a pH-sensitive sodium-dependent phosphate II transporter. SLC34A2 has been shown to be upregulated in breast and ovarian cancers and it is suggested that SLC34A2 is involved in the process of carcinogenesis, making it an attractive target in therapeutic strategies and also as a diagnostic biomarker. Although other antigens were found, and their sequences identified, all were unknown and not found in the databases. RT-PCR analysis of the Duke’s B colon cancer cell line SW480 showed consistent expression of BCP-20. Although, expression of SSX2, NY-ESO-1, TSP50, HAGE and RAGE were detected, the data was not easily reproductible. Further optimisation of the PCR conditions and primer pairs would be necessary to confirm these findings. We hope in the future we can disern the role of these antigens in the inflammatory immune responses associated with reactive Dukes’ B colon cancer which would help us better understand the mechanisms which underlie effective anti-tumour responses post-surgery. It may also be that RPL37A is a biomarker for patient survival in colorectal cancer and this would be worthy of further investigation. III DEDICATIONS In memory of my beloved grandmother who always believed in me and inspired me to pursue my dreams no matter how impossible they seemed. To Dr Barbara Guinn who is a true inspiration as a scientist, a role model and a friend. With a never-ending enthusiasm, positive energy and support she turned this research project into an unforgettable journey. Thank you for every moment. To my best friends and colleagues Emma and Kay who stood by me at all times. Thank you for always encouraging me to keep going when science and faith would fail. To Simon who reminded me that it is possible to have it all. To all my friends and colleagues from the University of Bedfordshire. Thank you for your support and friendship throughout my research degree. IV QUOTE “Around here, however, we don’t look backwards for very long. We keep moving forward, opening up new doors and doing new things… and curiosity keeps leading us down new paths.” - Walt Disney V PUBLICATIONS TO DATE Peer-reviewed publications 1. Hofmann, S., Khan, G., Boncheva, V., Greiner, J. & Guinn, B.A. (2014) Vaccination against myeloid leukaemias using newly defined antigens. In Press, Cancer Immunology Immunotherapy. (Ed. by Rees, R.) Oxford University Press. 2. Liberante, F.G., Pellagatti, A., Boncheva, V., Bowen, D.T., Mills, K.I., Boultwood, J. & Guinn, B.A. (2013). High and low, but not intermediate, PRAME expression levels are poor prognostic markers in myelodysplastic syndrome at disease presentation. British Journal of Haematology, 162, 282-284. 3. Boncheva, V., Bonney, S.A., Brooks, S.E., Tangney, M., O’Sullivan, G., Mirnezami, A. & Guinn, B.A. (2013) New targets for the immunotherapy of colon cancer – does reactive disease hold the answer? Cancer Gene Therapy, 20, 157-168. Conference presentations - Abstracts 1. Bonney, S.A., Brooks, S.E., Boncheva, V., Sigurdardottir, D., Tangney, M., Li, D., Patel, S., Pulford, K., Banham, A.H., Rammensee, H.G., O’Sullivan, G., Mirnezami, A. & Guinn, B.A. (2012) The identification of tumour-specific T cell populations recognized by colon cancer patients using the pMHC array. Orl presentation at the International Society for the Cell and Gene Therapy of Cancer annual conference, Singapore. 2. Khan, G., Boncheva, V., Mirnezami, A. & Guinn, B.A. (2013) Characterisation of SSX2 and SSX2IP expression in colon cancer. Poster presentation at the National Cancer Research Institute Annual Meeting. VI LIST OF CONTENTS ABSTRACT ........................................................................................................................................... II DEDICATIONS .................................................................................................................................... IV QUOTE .................................................................................................................................................. V PUBLICATIONS TO DATE ................................................................................................................ VI LIST OF CONTENTS .......................................................................................................................... VII LIST OF FIGURES ............................................................................................................................... IX LIST OF TABLES ................................................................................................................................. X ACKNOWLEDGEMENTS .................................................................................................................. XI ABBREVIATIONS .............................................................................................................................. XII CHAPTER 1 Introduction ............................................................................................................. 13 1.1 The Global Challenge ................................................................................................... 13 1.2 Colorectal Cancer (CRC).............................................................................................. 14 1.3 The Search for Tumour Antigens Specific for CRC .................................................... 18 1.4 Tumour profiling in patients with CRC ........................................................................ 26 1.5 Potential biomarkers for discerning reactive from non-reactive disease ...................... 31 1.6 Potential biomarkers for survival prognosis ................................................................. 32 1.7 Potential biomarkers for discerning aggressive from non-aggressive disease............. 33 1.8 Potential immunotherapeutic targets ........................................................................... 34 1.9 Non-reactive disease – overcoming the underlying issues ........................................... 34 1.10 Infiltrating T lymphocytes ............................................................................................ 36 1.11 Intrinsically disordered proteins (IDPs) ...................................................................... 39 1.12 Study Aims ................................................................................................................... 42 CHAPTER 2 Materials and Methods .......................................................................................... 43 2.1 First round of primary immunoscreening of a testes cDNA library (SEREX) ............. 43 2.2 Determination of serum reactivity and testis cDNA library efficiency ........................ 49 2.3 Construction of a new testis cDNA library................................................................... 53 2.4 Second round of primary immunoscreening with SEREX – optimised protocol ......... 56 2.5 Secondary
Load more

Recommended publications
  • Identifizierung Und Charakterisierung Von T-Zell-Definierten Antigenen
    Identifizierung und Charakterisierung von Zielantigenen alloreaktiver zytotoxischer T-Zellen mittels cDNA-Bank-Expressionsklonierung in akuten myeloischen Leukämien Dissertation zur Erlangung des Grades Doktor der Naturwissenschaften am Fachbereich Biologie der Johannes Gutenberg-Universität Mainz Sabine Domning Mainz, 2012 Fachbereich Biologie der Johannes Gutenberg-Universität Mainz Dekan: 1.Berichterstatter: 2.Berichterstatter: Tag der mündlichen Prüfung: ZUSAMMENFASSUNG Zusammenfassung Allogene hämatopoetische Stammzelltransplantationen (HSZTs) werden insbesondere zur Behandlung von Patienten mit Hochrisiko-Leukämien durchgeführt. Dabei bewirken T- Zellreaktionen gegen Minorhistokompatibilitätsantigene (mHAgs) sowohl den therapeutisch erwünschten graft-versus-leukemia (GvL)-Effekt als auch die schädigende graft-versus-host (GvH)- Erkrankung. Für die Identifizierung neuer mHAgs mittels des T-Zell-basierten cDNA- Expressionsscreenings waren leukämiereaktive T-Zellpopulationen durch Stimulation naïver CD8+- T-Lymphozyten gesunder HLA-Klasse I-identischer Buffy Coat-Spender mit Leukämiezellen von Patienten mit akuter myeloischer Leukämie (AML) generiert worden (Albrecht et al., Cancer Immunol. Immunother. 60:235, 2011). Im Rahmen der vorliegenden Arbeit wurde mit diesen im AML-Modell des Patienten MZ529 das mHAg CYBA-72Y identifiziert. Es resultiert aus einem bekannten Einzelnukleotidpolymorphismus (rs4673: CYBA-242T/C) des Gens CYBA (kodierend für Cytochrom b-245 α-Polypeptid; syn.: p22phox), der zu einem Austausch von Tyrosin (Y) zu Histidin (H) an Aminosäureposition 72 führt. Das mHAg wurde von T-Lymphozyten sowohl in Assoziation mit HLA-B*15:01 als auch mit HLA-B*15:07 erkannt. Eine allogene T-Zellantwort gegen CYBA-72Y wurde in einem weiteren AML-Modell (MZ987) beobachtet, die ebenso wie in dem AML-Modell MZ529 polyklonal war. Insgesamt konnte bei drei von fünf getesteten HLA-B*15:01-positiven Buffy Coat-Spendern, die homozygot für CYBA-72H (H/H) waren, eine CYBA-72Y-spezifische T- Zellantwort generiert werden.
    [Show full text]
  • Découverte De Nouveaux Transcrits De Fusion Dans Des Tumeurs
    Découverte de nouveaux transcrits de fusion dans des tumeurs pédiatriques en rechute et caractérisation fonctionnelle d’un nouvel oncogène LMO3-BORCS5 Celia Dupain Jourda To cite this version: Celia Dupain Jourda. Découverte de nouveaux transcrits de fusion dans des tumeurs pédiatriques en rechute et caractérisation fonctionnelle d’un nouvel oncogène LMO3-BORCS5. Cancer. Université Paris Saclay (COmUE), 2018. Français. NNT : 2018SACLS310. tel-02426230 HAL Id: tel-02426230 https://tel.archives-ouvertes.fr/tel-02426230 Submitted on 2 Jan 2020 HAL is a multi-disciplinary open access L’archive ouverte pluridisciplinaire HAL, est archive for the deposit and dissemination of sci- destinée au dépôt et à la diffusion de documents entific research documents, whether they are pub- scientifiques de niveau recherche, publiés ou non, lished or not. The documents may come from émanant des établissements d’enseignement et de teaching and research institutions in France or recherche français ou étrangers, des laboratoires abroad, or from public or private research centers. publics ou privés. Découverte de nouveaux transcrits de fusion dans des tumeurs pédiatriques en rechute et caractérisation fonctionnelle d’un nouvel oncogène LMO3-BORCS5 NNT: 2018SACLS310 NNT: Thèse de doctorat de l'Université Paris-Saclay préparée à l’Université Paris-Sud École doctorale n°568 : Signalisations et réseaux intégratifs en biologie (Biosigne) Spécialité de doctorat: Aspects moléculaires et cellulaires de la biologie Thèse présentée et soutenue à Villejuif, le 16 octobre
    [Show full text]
  • Transcriptional Expression of Zics As an Independent Indicator of Survival in Gliomas Zhaocheng Han, Jingnan Jia, Yangting Lv, Rongyanqi Wang & Kegang Cao*
    www.nature.com/scientificreports OPEN Transcriptional expression of ZICs as an independent indicator of survival in gliomas Zhaocheng Han, Jingnan Jia, Yangting Lv, Rongyanqi Wang & Kegang Cao* The functional signifcance of the zinc-fnger of the cerebellum (ZIC) gene family in gliomas remains to be elucidated. Clinical data from patients with gliomas, containing expression levels of ZIC genes, were extracted from CCLE, GEPIA2 and The Human Protein Atlas (HPA). Univariate survival analysis adjusted by Cox regression via OncoLnc was used to determine the prognostic signifcance of ZIC expression. We used cBioPortal to explore the correlation between gene mutations and overall survival (OS). ZIC expression was found to be related to immune cell infltration in gliomas via TIMER analysis. GO term and KEGG pathway enrichment analyzes were performed with Metascape. PPI networks were constructed using STRING. The expression levels of ZIC1/3/4/5 in gliomas were signifcantly diferent from those in normal samples. High expression levels of ZIC1/5 were associated with poor OS in brain low-grade glioma (LGG) patients, while low ZIC3 expression combined was related to favorable OS in glioblastoma multiforme (GBM). ZIC alterations were associated with poor prognosis in LGG patients and related to favorable prognosis in GBM patients. We observed that the expression of ZICs was related to immune cell infltration in glioma patients. ZICs were enriched in several pathways and biological processes involving Neuroactive ligand-receptor interaction (hsa04080). The PPI network revealed that some proteins coexpressed with ZICs played a role in the pathogenesis of gliomas. Diferences in the expression levels of ZIC genes could provide a signifcant marker for predicting prognosis in gliomas.
    [Show full text]
  • Genome-Wide Analysis of Cancer/Testis Gene Expression
    Genome-wide analysis of cancer/testis gene expression Oliver Hofmanna,b,1, Otavia L. Caballeroc, Brian J. Stevensond,e, Yao-Tseng Chenf, Tzeela Cohenc, Ramon Chuac, Christopher A. Maherb, Sumir Panjib, Ulf Schaeferb, Adele Krugerb, Minna Lehvaslaihob, Piero Carnincig,h, Yoshihide Hayashizakig,h, C. Victor Jongeneeld,e, Andrew J. G. Simpsonc, Lloyd J. Oldc,1, and Winston Hidea,b aDepartment of Biostatistics, Harvard School of Public Health, 655 Huntington Avenue, SPH2, 4th Floor, Boston, MA 02115; bSouth African National Bioinformatics Institute, University of the Western Cape, Private Bag X17, Bellville 7535, South Africa; cLudwig Institute for Cancer Research, New York Branch at Memorial Sloan-Kettering Cancer Center, 1275 York Avenue, New York, NY 10021; dLudwig Institute for Cancer Research, Lausanne Branch, 1015 Lausanne, Switzerland; eSwiss Institute of Bioinformatics, 1015 Lausanne, Switzerland; fWeill Medical College of Cornell University, 1300 York Avenue, New York, NY 10021; gGenome Exploration Research Group (Genome Network Project Core Group), RIKEN Genomic Sciences Center (GSC), RIKEN Yokohama Institute, 1-7-22 Suehiro-cho, Tsurumi-ku, Yokohama, Kanagawa, 230-0045, Japan; and hGenome Science Laboratory, Discovery Research Institute, RIKEN Wako Institute, 2-1 Hirosawa, Wako, Saitama, 3510198, Japan Contributed by Lloyd J. Old, October 28, 2008 (sent for review June 6, 2008) Cancer/Testis (CT) genes, normally expressed in germ line cells but expression profile information frequently limited to the original also activated in a wide range of cancer types, often encode defining articles. In some cases, e.g., ACRBP, the original antigens that are immunogenic in cancer patients, and present CT-restricted expression in normal tissues could not be con- potential for use as biomarkers and targets for immunotherapy.
    [Show full text]
  • Characterisation of the Expression of Tumour Antigens and Biomarkers in Myeloid Leukaemia and Ovarian Cancer
    Title : Characterisation of the expression of tumour antigens and biomarkers in myeloid leukaemia and ovarian cancer Name : Ghazala Naz Khan This is a digitised version of a dissertation submitted to the University of Bedfordshire. It is available to view only. This item is subject to copyright. Characterisation of the Expression of Tumour Antigens and Biomarkers in Myeloid Leukaemia and Ovarian Cancer By Ghazala Naz Khan PhD A thesis submitted to the Faculty of Creative Arts, Technologies and Science, University of Bedfordshire in fulfilment of the requirements for the degree of Doctor of Philosophy December 2016 Abstract Acute myeloid leukaemia (AML) and ovarian cancer (OVC) are two difficult to treat cancers. AML is often treatable however minimal residual disease (MRD) endures such that many patients who achieve remission eventually relapse and succumb to the disease. OVC affects approximately 7000 women in the U.K. every year. It can occur at any age but is most common after menopause. Diagnosis at an early stage of disease greatly improves the chances of survival however, patients tend to be diagnosed in the later stages of disease when treatment is often less effective. Immunotherapy has the potential to reduce MRD and delay or prevent relapse. In order for immunotherapy to work, tumour antigens need to be identified and characterised so they can be effectively targeted. Personalised treatments require the identification of biomarkers, for disease detection and confirmation, as well as to provide an indication of best treatment and the prediction of survival. PASD1 has been found to be frequently expressed in haematological malignancies and I wanted to determine if there was a correlation between the presence of antigen-specific T cells in the periphery of patients with AML and PASD1 protein expression in the leukaemic cells.
    [Show full text]
  • Number 9 September 2013
    Atlas of Genetics and Cytogenetics in Oncology and Haematology OPEN ACCESS JOURNAL INIST -CNRS Volume 17 - Number 9 September 2013 The PDF version of the Atlas of Genetics and Cytogenetics in Oncology and Haematology is a reissue of the original articles published in collaboration with the Institute for Scientific and Technical Information (INstitut de l’Information Scientifique et Technique - INIST) of the French National Center for Scientific Research (CNRS) on its electronic publishing platform I-Revues. Online and PDF versions of the Atlas of Genetics and Cytogenetics in Oncology and Haematology are hosted by INIST-CNRS. Atlas of Genetics and Cytogenetics in Oncology and Haematology OPEN ACCESS JOURNAL INIST -CNRS Scope The Atlas of Genetics and Cytogenetics in Oncology and Haematology is a peer reviewed on-line journal in open access, devoted to genes, cytogenetics, and clinical entities in cancer, and cancer-prone diseases. It presents structured review articles ("cards") on genes, leukaemias, solid tumours, cancer-prone diseases, more traditional review articles on these and also on surrounding topics ("deep insights"), case reports in hematology, and educational items in the various related topics for students in Medicine and in Sciences. Editorial correspondance Jean-Loup Huret Genetics, Department of Medical Information, University Hospital F-86021 Poitiers, France tel +33 5 49 44 45 46 or +33 5 49 45 47 67 [email protected] or [email protected] Staff Mohammad Ahmad, Mélanie Arsaban, Marie-Christine Jacquemot-Perbal, Vanessa Le Berre, Anne Malo, Carol Moreau, Catherine Morel-Pair, Laurent Rassinoux, Alain Zasadzinski. Philippe Dessen is the Database Director, and Alain Bernheim the Chairman of the on-line version (Gustave Roussy Institute – Villejuif – France).
    [Show full text]
  • Genetics of Migration Timing in Bar-Tailed Godwits
    Copyright is owned by the Author of the thesis. Permission is given for a copy to be downloaded by an individual for the purpose of research and private study only. The thesis may not be reproduced elsewhere without the permission of the Author. Genetics of migration timing in bar-tailed godwits A thesis presented in partial fulfilment of the requirements for the degree of Doctor of Philosophy in Zoology at Massey University, Manawatū, New Zealand. Ángela María Parody Merino 2018 Dedicado a mis padres; vosotros me disteis alas para volar. Preface In this thesis, I used molecular and genomic techniques to give insight on the genetic elements behind migratory timing behaviour in a very suitable natural population bird system: The New Zealand godwit. This thesis is structured as a series of related and connected manuscripts with exception of the introduction, which provides the terminology, background and founding of my thesis: Each chapter is a stand-alone piece of work, therefore, there will be some unavoidable repetition between them and the introduction. This thesis contains three main chapters (see listed below), which are intended to be published. • Chapter 2: Parody-Merino A.M., Battley P.F., Verkuil Y. I., Conklin J.R., Prosdocimi F., Potter, M.A., Fidler A.E. Population genetic structure of New Zealand-overwintering bar-tailed godwits (Limosa lapponica baueri) in relation to breeding geography and migration timing. • Chapter 3: Parody-Merino A.M., Battley P.F., Conklin J.R., Fidler A.E. No evidence for an association between Clock gene allelic variation and migration timing in a long-distance migratory shorebird (Limosa lapponica baueri).
    [Show full text]
  • Characterization of X-Linked SNP Genotypic Variation in Globally Distributed Human Populations Genome Biology 2010, 11:R10
    Casto et al. Genome Biology 2010, 11:R10 http://genomebiology.com/2010/11/1/R10 RESEARCH Open Access CharacterizationResearch of X-Linked SNP genotypic variation in globally distributed human populations Amanda M Casto*1, Jun Z Li2, Devin Absher3, Richard Myers3, Sohini Ramachandran4 and Marcus W Feldman5 HumanAnhumanulation analysis structurepopulationsX-linked of X-linked variationand provides de geneticmographic insights variation patterns. into in pop- Abstract Background: The transmission pattern of the human X chromosome reduces its population size relative to the autosomes, subjects it to disproportionate influence by female demography, and leaves X-linked mutations exposed to selection in males. As a result, the analysis of X-linked genomic variation can provide insights into the influence of demography and selection on the human genome. Here we characterize the genomic variation represented by 16,297 X-linked SNPs genotyped in the CEPH human genome diversity project samples. Results: We found that X chromosomes tend to be more differentiated between human populations than autosomes, with several notable exceptions. Comparisons between genetically distant populations also showed an excess of X- linked SNPs with large allele frequency differences. Combining information about these SNPs with results from tests designed to detect selective sweeps, we identified two regions that were clear outliers from the rest of the X chromosome for haplotype structure and allele frequency distribution. We were also able to more precisely define the geographical extent of some previously described X-linked selective sweeps. Conclusions: The relationship between male and female demographic histories is likely to be complex as evidence supporting different conclusions can be found in the same dataset.
    [Show full text]
  • PASD1: a Promising Target for The
    Khan et al., J Genet Syndr Gene Ther 2013, 4:9 Genetic Syndromes & Gene Therapy http://dx.doi.org/10.4172/2157-7412.1000186 Review Article Open Access PASD1: A Promising Target for the Immunotherapy of Haematological Malignancies Ghazala Khan1, Frances Denniss2, Ken I Mills3, Karen Pulford4 and Barbara-ann Guinn1,2* 1Department of Life Sciences, University of Bedfordshire, Park Square, Luton, LU1 3JU, UK 2Department of Haematological Medicine, The Rayne Institute, King’s College London School of Medicine SE5 9NU, UK 3Centre for Cancer Research and Cell Biology, Queens University Belfast, Belfast, BT9 7BL, UK 4Nuffield Division of Clinical Laboratory Sciences, University of Oxford, John Radcliffe Hospital, Oxford, OX3 9DU, UK Abstract In general, there is a lack of good immunotherapy targets within the spectrum of haematological malignancies. However haematopoietic stem cell transplants and continuing antigen discovery have allowed further insight into how further improvements in outcomes for patients might be achieved. Most patients with haematological malignancies can be treated with conventional therapies such as radio- and chemotherapy and will attain first remission. However the removal of residual diseased cells is essential to prevent relapse and its associated high mortality. PASD1 is one of the most tissue restricted cancer-testis (CT) antigens with expression limited to primary spermatagonia in healthy tissue. However, characterisation of PASD1 expression in cancers has been predominantly focussed on haematological malignancies where the inappropriate expression of PASD1 was first identified. PASD1 has one of the highest frequencies of expression of all CT antigens in acute myeloid leukaemia, with some suggestion of its role as a biomarker in diffuse large B-cell lymphoma.
    [Show full text]
  • Effect of 5-Ht on Cell Behaviour
    Differential expression of C/T antigens and 5- HT receptors in Normotensive and Pre- eclamptic placentae, and their role in trophoblast invasion. A Thesis Submitted by Anushuya Priyadarshini Tamang To The Nottingham Trent University In requirement for the degree of Doctor of Philosophy April 2015 Copyright statement This work is the intellectual property of the author, and may also be owned by the research sponsor(s) and/or Nottingham Trent University. You may copy up to 5% of this work for private study, or personal, non-commercial research. Any re-use of the information contained within this document should be fully referenced, quoting the author, title, university, degree level and pagination. Queries or requests for any other use, or if a more substantial copy is required, should be directed in the first instance to the author. i Acknowledgements First of all I would like to thank my Director of Study, Dr.Shiva Sivasubramaniam for believing in me and giving me an opportunity to work on this project. I am grateful for your constant support, encouragement and some minor stress. I would also like to thank my supervisors Dr Morgan Geoffrey Mathieu and Professor Robert Rees for their valuable insight into this project. Next, I would like to thank my technical support team, Mike, Jackey and Dan, for always helping me with a smile and turning a blind eye to everything that went missing from their lab. (You know where to find them.) I am indebted to Gordan, for helping me through the painfully long hours of microscope settings and never complaining about it.
    [Show full text]
  • UNIVERSITY of CALIFORNIA SANTA CRUZ the CELLULAR CONSEQUENCES of PASD1 EXPRESSION in HUMAN CANCER a Thesis Submitted in Partial
    UNIVERSITY OF CALIFORNIA SANTA CRUZ THE CELLULAR CONSEQUENCES OF PASD1 EXPRESSION IN HUMAN CANCER A thesis submitted in partial satisfaction of the requirements for the degree of MASTER OF ARTS in MOLECULAR, CELL, AND DEVELOPMENTAL BIOLOGY by Ashley M. Kern March 2016 The Thesis of Ashley M. Kern is approved: ____________________________________ Carrie Partch, Ph.D., chair ____________________________________ Seth Rubin, Ph.D. ____________________________________ Grant Hartzog, Ph.D. _______________________________ Tyrus Miller Vice-Provost and Dean of Graduate Studies ii Table of Contents Title Page I Table of Contents iii Abstract v Acknowledgements vi Figure/Experimental Contributions vii Introduction 1 Results 23 Discussion 39 Experimental Procedures 46 Figures 3- 45 Figure 1 3 Figure 2 6 Figure 3 12 Figure 4 14 Figure 5 15 Figure 6 22 Figure 7 27 Figure 8 30 Figure 9 36 Figure 10 38 Figure 11 42 iii Figure 12 45 References 48 iv ABSTRACT The Cellular Consequences of PASD1 Expression in Human Cancer Ashley M. Kern There are nearly 1,000 human proteins that are expressed only in the germline. Cancer /Testis antigens (CT antigens) represent a subset of germline-specific proteins that become reactivated in somatic cells that have undergone oncogenic transformation. Human PAS Domain containing protein 1 (PASD1) is an X-linked CT Antigen. In 2015, the Partch Laboratory established that PASD1 prevents the core circadian transcription factor, CLOCK:BMAL1, from rhythmically transcribing its target genes to suppress circadian rhythms after it becomes upregulated in cancer cells. In addition to this striking phenotype, PASD1 also promotes mitotic defects that could favor tumor progression. These phenotypes may go hand in hand; over 43% of the mammalian genome, including many cell cycle genes, is under circadian control.
    [Show full text]
  • Avaliação Do Nível De Expressão De MAGE A1 E BORIS E Do Perfil De Metilação Em Carcinoma De Células Escamosas Bucal
    UNIVERSIDADE FEDERAL DE MINAS GERAIS PROGRAMA DE PÓS-GRADUAÇÃO EM EM CIÊNCIAS BIOLÓGICAS: FARMACOLOGIA, BIOQUÍMICA E MOLECULAR Avaliação do nível de expressão de MAGE A1 e BORIS e do perfil de metilação em carcinoma de células escamosas bucal CLÁUDIA MARIA PEREIRA Belo Horizonte 2011 Avaliação do nível de expressão de MAGE A1 e BORIS e do perfil de metilação em carcinoma de células escamosas bucal CLÁUDIA MARIA PEREIRA Tese apresentada ao Programa de Pós- Graduação em Ciências Biológicas: Farmacologia, Bioquímica e Molecular da Universidade Federal de Minas Gerais como requisito parcial para a obtenção do título de Doutora em Farmacologia, Bioquímica e Molecular Orientador: Prof. Dr. Ricardo Santiago Gomez Belo Horizonte Minas Gerais 2011 “Cuidado com o que você pede e deseja; Você pode conseguir” Autor Desconhecido DEDICATÓRIA Dedico a minha dissertação primeiramente a Deus e à Nossa Senhora, que estiveram sempre ao meu lado, me direcionando e protegendo durante toda esta jornada. Dedico também aos meus pais Eure e Célia, pelo apoio, amor e carinho. Dedico aos meus irmãos (Antônio, Fernando, Carlos, Roberto e Caio), aos meus sobrinhos (Ludmila, Angelina, Matheus, Mariana, Pedro, Marcos e Giovanna) e às minhas cunhadas (Carla, Inês, Mônica e Glaura), que mesmo distantes, estiveram sempre presentes me apoiando e me incentivando. Dedico ao meu orientador Prof. Dr. Ricardo Santiago Gomez por confiar e acreditar em mim durante estes anos. AGRADECIMENTOS Agradeço ao meu orientador Dr. Ricardo Santiago Gomez por ter me acompanhado desde o mestrado, por ter me dado a oportunidade de fazer parte de sua equipe em meu doutorado e por permitir meu crescimento profissional e pessoal.
    [Show full text]