• Abstract and Figures
• Public Full-text

Comparative oncology research in Australia: Establishing the Australian Veterinary Cancer Biobank to validate canine mammary tumours as a model for human breast cancer A thesis submitted in fulfilment of the requirements for the degree of Doctor of Philosophy Kristi Maree Milley B. App. Sci (Laboratory Medicine) Hons. RMIT University School of Health and Biomedical Sciences College of Science Engineering and Health RMIT University February 2018 ii Declaration I certify that except where due acknowledgement has been made, the work is that of the author alone; the work has not been submitted previously, in whole or in part, to qualify for any other academic award; the content of the thesis is the result of work which has been carried out since the official commencement date of the approved research program; any editorial work, paid or unpaid, carried out by a third party is acknowledged; and, ethics procedures and guidelines have been followed. I acknowledge the support I have received for my research through the provision of an Australian Government Research Training Program Scholarship. Kristi Maree Milley 30th January 2018 iii Acknowledgements “A mentor is someone who sees more talent and ability within you, than you see in yourself, and helps bring it out of you” – Bob Proctor My biggest acknowledgement is to my supervisors Associate Professor Janine Danks and Associate Professor Samantha Richardson. Janine ‘recruited’ me into Honours as an undergraduate and spent a full year promoting the virtues of research. She has gone above and beyond her role as my supervisor. She has been my mentor and supporter for the past nine years. She has taught me the value of open collaboration, supporting other women in research and how to recognise the value in my own work. When I could not see my way out of my situation and the darkness around me she patiently supported me, she even found me a job and endlessly encouraged me to continue. I would like to thank my comrades in arms Suzan Sam and Damian D’Souza for their company and conversation through many long days, late nights and weekends. Lastly, during the past eight years, my family has supported me emotionally and financially and asked how my PhD was going no less than 1000 times. So, I would like to thank them for their patience and love. iv Contents Declaration ................................................................................................. iii Acknowledgements.................................................................................... iv Contents .....................................................................................................v List of Figures .......................................................................................... xiii List of Tables ........................................................................................... xix List of Abbreviations ............................................................................... xxii Manuscripts relating to this thesis .......................................................... xxv Conferences relating to this thesis .............................................................1 Research summary: ....................................................................................2 CHAPTER 1: ................................................................................................7 INTRODUCTION ..........................................................................................7 1.Introduction:.............................................................................................9 1.1 Breast cancer .................................................................................. 9 1.2 Molecular subtypes of breast cancer ................................................10 1.2.1 Breast cancer and its classification ............................................10 1.2.2 Molecular subtyping of human breast cancer ..............................11 1.2.3 Mechanism of metastasis ..........................................................16 1.2.4 Molecular subtypes of breast cancer and metastasis ...................18 1.3 Comparative oncology: Animal models for breast cancer ...................20 1.4 Canine mammary tumours ..............................................................24 1.4.1 Classification and grading of CMTs ............................................24 1.4.2 Similarity of canine mammary tumours to human breast cancer ...25 1.4.3 Factors influencing normal mammary tissue and CMT.................29 1.5 Biobanking for breast cancer research .............................................31 1.6 Potential prognostic markers for CMT...............................................35 v 1.6.1 Runx-2 .....................................................................................37 1.6.2 Parathyroid hormone-related protein in breast cancer .................38 1.6.3 GATA-3....................................................................................49 1.7 Aims and scope of this study ...........................................................51 CHAPTER 2: .............................................................................................. 53 Materials and methods .............................................................................. 53 2.Materials and methods ........................................................................... 55 2.1 Biobank design and pilot project.......................................................55 2.2 Participant recruitment ....................................................................56 2.3 Sample collection and registration ....................................................60 2.4 Data entry and storage ....................................................................68 2.5 Sample processing .........................................................................70 2.5.1 Macroscopic description and dissection......................................72 2.5.2 Fixed tissue specimen processing..............................................74 2.6 DogMATIC specimen histopathological diagnosis .............................77 2.6.1 Other specimen histopathological diagnosis ...............................77 2.7 Evaluating sample collection feasibility and sustainability of the AVCBB 77 2.7.1 DogMATIC kit validation ............................................................78 2.7.2 DogMATIC kit success – The view of biobank network clinics ......79 2.7.3 Sample collection rate ...............................................................80 2.7.4 Transit time ..............................................................................80 2.7.5 Testing RNA quality and quantity of DogMATIC samples.............80 vi 2.7.6 Effect of transit time on RNA quality ...........................................81 2.7.7 Follow-up data collection ...........................................................82 2.8 Tissue microarray construction ........................................................83 2.9 Immunohistochemistry ....................................................................85 2.9.1 Subtyping malignant CMTs........................................................85 2.9.2 Prognostic marker study............................................................89 2.10 Mouse osteosarcoma study ..........................................................90 2.10.1 Mouse OS IHC ......................................................................92 2.11 Image scoring and analysis ..........................................................94 2.11.1 Scoring and image analysis ....................................................94 2.12 Statistical analysis ..................................................................... 107 2.12.1 Animal demographics and survival ........................................ 107 2.12.2 IHC analysis ........................................................................ 108 CHAPTER 3: ............................................................................................ 111 Establishing a canine mammary tumour (CMT) Biobank for Comparative Oncology Research................................................................................. 111 3.Establishing a canine mammary tumour biobank for comparative oncology research .................................................................................. 113 3.1 Introduction .................................................................................. 113 3.2 Results - AVCBB pilot study .......................................................... 116 3.2.1 Biobank network ..................................................................... 116 3.2.2 Infrastructure and materials created ......................................... 117 3.2.3 The DogMATIC kit produces a moderate sample size ............... 125 3.2.4 DogMATIC kit survey analysis ................................................. 127 vii 3.2.5 DogMATIC kit provides intact, quality total RNA ........................ 129 3.2.6 DogMATIC transit time ............................................................ 133 3.2.7 Potential effect of transit on RNA concentration ........................ 133 3.2.8 Summary of pilot study collection ............................................. 134 3.2.9 Overall collection period- DogMATIC kits.................................. 136 3.2.10 Summary overall collection period, fixed samples .................. 139 3.2.11 Animal follow-up, data loss ................................................... 141 3.3 Discussion...................................................................................

Load more

  634

Copy Link