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Genetics of Congenital Heart Diseases

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Genetics of Congenital Heart Diseases PLEASE TYPE THE UNIVERSITY OF NEW SOUTH WALES Thesis/Dissertation Sheet Surname or Family name: Moradi Marjaneh First name: Mahdi Other name/s: Abbreviation for degree as given in the University calendar: PhD School: St Vincent's Clinical School Faculty: Medicine Title: Genetics of Congenital Heart Diseases Abstract 350 words maximum: (PLEASE TYPE) Development of the cardiac atrial septum involves complex morphogenetic processes including programmed cell growth and death. Secundum atrial s eptal d efect ( ASDII) an d p atent f oramen o vale ( PFO) ar e co mmon at rial s eptal an omalies as sociated with n umerous p athologies including s troke. D ata from studies i n hum ans a nd mouse s uggest t hat P FO a nd A SDII e xist i n a n a natomical c ontinuum of septal dysmorphogenesis with a common genetic basis. Analysis of quantitative trait loci (QTL) and genome technology form a powerful approach to understand genetic complexity underpinning common disease. A previous study o f inbred mice mapped QTL for quantitative anatomical atrial s eptal p arameters correlating with PFO, including flap valve length (FVL) and foramen ovale width (FOW). Here, we explore an advanced intercross line (AIL) for confirmation and fine mapping of t hese Q TL. An A IL be tween pa rental s trains QSi5 a nd 129T2/SvEms, s howing e xtreme va lues f or F VL a nd PFO, w as established ov er 1 4 g enerations. L inkage a nalysis us ing 141 s ingle nuc leotide p olymorphism m arkers f ocused on 6 s ignificant a nd on e suggestive QTL regions for FVL or FOW found previously, and we also sought QTL for heart weight (HW) normalized to body weight (BW). Virtually all QTL were confirmed and refined and many new QTL were discovered, with analysis of PFO as a binary trait providing strong support. QTL were not explained by HW/BW differences between parental strains. The overlap between FVL and FOW QTL was striking, indicating many QTL affect processes relevant to both septum primum and septum secundum. This study provides a high-resolution picture of genetic c omplexity unde rpinning a trial s eptal va riation i n t he m ouse a nd pr edicts i nvolvement of pot entially hundr eds of r isk variants i n humans. Subsequently, high throughput sequencing of the whole genomes of the parental lines and analysis of HapMap datasets as a validation method, in addition to applying a multi-step filtering criteria resulted in identification of gene variants underlying QTL, and candidates for involvement in phenotypic variation based on the predicted impact of sequence changes. Mutations in multiple members of the evolutionarily conserved cardiac transcription factor network, including GATA4, cause or predispose to ASDII and PFO. Here, we assessed whether the most prevalent variant of the GATA4 gene, S377G, was significantly associated with PFO or ASD. Ou r a nalysis o f world i ndigenous populations s howed t hat G ATA4 S 377G was l argely Caucasian-specific, and s o s ubjects were restricted to those of Caucasian descent. To select for patients with larger PFO, we limited our analysis to those with cryptogenic stroke in which PFO was a subsequent finding. In an initial study of Australian subjects, a weak association between GATA4 S377G and PFO/Stroke was observed. However, in a follow up study of German Caucasians no association was found with either PFO or ASD. Analysis of combined Australian and German data confirmed the lack of association. Thus, the common GATA4 variant S377G is likely to be relatively benign in terms of its participation in CHD and PFO/Stroke. Declaration relating to disposition of project thesis/dissertation I hereby grant to the University of New South Wales or its agents the right to archive and to make available my thesis or dissertation in whole or in part in the University libraries in all forms of media, now or here after known, subject to the provisions of the Copyright Act 1968. I retain all property rights, such as patent rights. I also retain the right to use in future works (such as articles or books) all or part of this thesis or dissertation. I also authorise University Microfilms to use the 350 word abstract of my thesis in Dissertation Abstracts International (this is applicable to doctoral theses only). …………………………………………………………… ……………………………………..……………… ……….……………………...…….… Signature Witness Date The University recognises that there may be exceptional circumstances requiring restrictions on copying or conditions on use. Requests for restriction for a period of up to 2 years must be made in writing. Requests for a longer period of restriction may be considered in exceptional circumstances and require the approval of the Dean of Graduate Research. FOR OFFICE USE ONLY Date of completion of requirements for Award: THIS SHEET IS TO BE GLUED TO THE INSIDE FRONT COVER OF THE THESIS COPYRIGHT STATEMENT ‘I hereby grant the University of New South Wales or its agents the right to archive and to make available my thesis or dissertation in whole or part in the University libraries in all forms of media, now or here after known, subject to the provisions of the Copyright Act 1968. I retain all proprietary rights, such as patent rights. I also retain the right to use in future works (such as articles or books) all or part of this thesis or dissertation. I also authorise University Microfilms to use the 350 word abstract of my thesis in Dissertation Abstract International (this is applicable to doctoral theses only). I have either used no substantial portions of copyright material in my thesis or I have obtained permission to use copyright material; where permission has not been granted I have applied/will apply for a partial restriction of the digital copy of my thesis or dissertation.' Signed ……………………………………………........................... Date ……………………………………………........................... AUTHENTICITY STATEMENT ‘I certify that the Library deposit digital copy is a direct equivalent of the final officially approved version of my thesis. No emendation of content has occurred and if there are any minor variations in formatting, they are the result of the conversion to digital format.’ Signed ……………………………………………........................... Date ……………………………………………........................... ORIGINALITY STATEMENT ‘I hereby declare that this submission is my own work and to the best of my knowledge it contains no materials previously published or written by another person, or substantial proportions of material which have been accepted for the award of any other degree or diploma at UNSW or any other educational institution, except where due acknowledgement is made in the thesis. Any contribution made to the research by others, with whom I have worked at UNSW or elsewhere, is explicitly acknowledged in the thesis. I also declare that the intellectual content of this thesis is the product of my own work, except to the extent that assistance from others in the project's design and conception or in style, presentation and linguistic expression is acknowledged.’ Signed …………………………………………….............. Date …………………………………………….............. Genetics of Congenital Heart Diseases MAHDI MORADI MARJANEH A thesis in fulfilment of the requirements for the degree of Doctor of Philosophy August, 2012 St Vincent's Clinical School University of New South Wales Dedication To my dear wife and parents. With my deepest love, gratitude, and respect. i Acknowledgment During t he c ourse of P hD, I ha ve be come i ndebted t o m any pe ople f or t heir g enerous support and help of many kinds. First and foremost I am grateful to my family in particular, my parents for their unconditional love, never-ending support, and encouragement during all of m y e ducational e ndeavours. I met my w ife, M adinah Y ate, w hen I w as do ing t he l ast semester of PhD. Since then my l ife has changed so much f or t he better. She has been a great emotional support and inspiration for me. Academically, my greatest debts are, of course, to my supervisor, Prof. Richard Harvey, and co-supervisor, A/Prof. Edwin Kirk, for all their guidance, support, help, and encouragement. I am eternally obliged to them for everything I learnt from them. I am grateful to all people in the Victor Chang Cardiac Research Institute (VCCRI) and, of course a bove a ll, members of H arvey l ab, w here I di d m y P hD, f or t heir he lp in v arious ways and friendship over the years. Especially, I would like to thank Dr. Naisana Asli for teaching me many lab techniques and helping me with any technical questions that I have had, Dr. Mirana Ramialison and Mrs. Tram Doan for teaching me and helping me with the bioinformatics techniques, and Dr. Gonzalo del Monte Nieto for teaching me t o dissect mouse hearts, and for providing ongoing help with the dissection of mouse atrial septums. I would l ike t o a ddress s pecial t hanks t o P rof. S ally D unwoodie, a m ember of m y P hD committee, for her precious feedbacks in the PhD reviews. There have been many others at VCCRI who h elped m e i n v arious w ays. I a m e specially g rateful to M r. A mirsalar Rashidianfar, D r. A shley Waar denberg, Ms. B ernice S tewart, P rof. B ob G raham, Mr . Brendan Lee, A/Prof. Catherine Suter, Ms. Dasha Syal, Dr. David Humphreys, Ms. Karen Brennan, Mr. Khai Do, Dr. Michael Swanton, Dr. Munira Xaymardan, Mr. Pardeep Dhiman, Mr. P aul Y oung, D r.
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