The University of Notre Dame Australia ResearchOnline@ND Theses 2018 Investigating the molecular pathology of Dupuytren’s disease Robert Pearce The University of Notre Dame Australia Follow this and additional works at: https://researchonline.nd.edu.au/theses Part of the Medicine and Health Sciences Commons COMMONWEALTH OF AUSTRALIA Copyright Regulations 1969 WARNING The material in this communication may be subject to copyright under the Act. Any further copying or communication of this material by you may be the subject of copyright protection under the Act. Do not remove this notice. Publication Details Pearce, R. (2018). Investigating the molecular pathology of Dupuytren’s disease (Doctor of Philosophy (College of Medicine)). University of Notre Dame Australia. https://researchonline.nd.edu.au/theses/209 This dissertation/thesis is brought to you by ResearchOnline@ND. It has been accepted for inclusion in Theses by an authorized administrator of ResearchOnline@ND. 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Investigating the Molecular Pathology of Dupuytren’s Disease Investigation of the Molecular Pathology of Dupuytren’s Disease Robert Pearce FRACS This thesis is presented for the degree of Doctor of Philosophy School of Medicine University of Notre Dame Australia (Fremantle) 2018 1 Contents Investigation of the Molecular Pathology of Dupuytren’s Disease ............................................................... 1 ABSTRACT .................................................................................................................................................. 7 ACKNOWLEDGEMENTS ........................................................................................................................ 10 CHAPTER 1 .............................................................................................................................................. 11 1.1 An overview of Dupuytren’s disease ................................................................................................ 11 1.2 Current clinical management of Dupuytren’s disease ....................................................................... 13 1.2.1 Prevention .................................................................................................................................. 13 1.3 Surgery to treat Dupuytren’s disease ................................................................................................. 13 1.3.1 Limited or partial fasciectomy.................................................................................................... 14 1.3.2 Dermofasciectomy ..................................................................................................................... 14 1.3.3 Z-Plasty repair ............................................................................................................................ 15 1.3.4 Amputation ................................................................................................................................. 15 1.3.5Percutaneous division of fibrous cords ........................................................................................ 16 1.4 Non-surgical treatment of Dupuytren’s disease ................................................................................ 16 1.4.1 Physical manipulation of Dupuytren’s contracture .................................................................... 16 1.4.2 Irradiation ................................................................................................................................... 17 1.4.3 Extracorporeal shockwave therapy ............................................................................................. 17 1.5 Pharmacological treatments of Dupuytren’s disease ......................................................................... 17 1.5.1 Intralesional steroid injection ..................................................................................................... 18 1.5.2 Intralesional collagenase injection ............................................................................................. 18 1.5.3 Intralesional gamma-interferon .................................................................................................. 18 1.6 Summary ........................................................................................................................................... 19 1.7 The Epidemiology of Dupuytren’s disease ....................................................................................... 20 1.7.1 Epidemiological evidence for a genetic component to Dupuytren’s disease. ............................. 21 1.8 The Molecular and cellular pathology of Dupuytren’s disease ......................................................... 22 1.8.1 The role of myofibroblasts in Dupuytren’s disease .................................................................... 22 1.8.2 The role of the immune system in Dupuytren’s disease ............................................................. 25 1.8.3 A role for vascular and innervation changes in Dupuytren’s disease ......................................... 26 1.9 Evidence for candidate susceptibility genes in Dupuytren’s disease ................................................ 27 1.10 Summary ......................................................................................................................................... 31 Hypothesis ................................................................................................................................................... 32 Aims ............................................................................................................................................................ 32 CHAPTER 2 .............................................................................................................................................. 33 2.1 Introduction ....................................................................................................................................... 33 Investigating the Molecular Pathology of Dupuytren’s Disease 2.2 Methods ............................................................................................................................................. 35 2.2.1 Ethics Approval .......................................................................................................................... 35 2.2.2 Participants ................................................................................................................................. 35 2.2.3 Saliva collection for genomic DNA isolation ............................................................................. 35 2.2.4 DNA isolation protocol .............................................................................................................. 36 2.2.5 DNA quantitation ....................................................................................................................... 36 2.2.6 Whole exome sequencing (WES) ............................................................................................... 36 2.2.7 Linkage analysis ......................................................................................................................... 38 2.2.8 Variant confirmation and prioritization ...................................................................................... 38 2.3 Results ............................................................................................................................................... 38 2.3.1 Genes identified using the SOLiD platform ............................................................................... 38 2.3.2 Genes identified using the Ion Proton sequencing platform ....................................................... 42 2.3.4 Genes identified using the Illumina platform ............................................................................. 50 2.3.5 Summary .................................................................................................................................... 50 2.4 Discussion ......................................................................................................................................... 51 2.4.1 Non-synonymous variants identified by WES in a family with Dupuytren’s disease ................ 51 2.4.2 Genes involved in angiogenesis ................................................................................................. 54 2.4.3 Genes with roles in inflammatory processes .............................................................................. 54 2.4.4 Candidate genes with neural functions ....................................................................................... 55 2.4.5 The absence of genes identified in previous studies ................................................................... 56 2.4.6 The advantages of using WES to identify causative mutations .................................................. 57 2.4.7 Limitations of using the WES approach ..................................................................................... 58 2.5 Summary ........................................................................................................................................... 61 CHAPTER 3 ............................................................................................................................................... 63 3.1 Introduction ......................................................................................................................................