Professor Doug Brooks 

Professor Doug Brooks is the leader of the Mechanisms in Cell Biology and Disease Research Group at the Sansom Institute for Health Sciences in the School of Pharmacy and Medical Science at the University of South Australia. He has over 25 years experience in medical research and is an NH&MRC funded Senior Research Fellow and Research Professor in Molecular Medicine. His initial research training was in Immunology with a focus on cancer research, involving the immunochemistry of cell surface antigens. For 24 years he worked in the Lysosomal Diseases Research Unit at the Women’s and Children’s Hospital, on a group of genetic diseases called lysosomal storage disorders. The Lysosomal Diseases Research Unit has been responsible for significant health outcomes for this group of disorders, with the development of strategies for early screening, diagnosis and treatment. This research reflects his strong interest in lysosomal cell biology and a desire to develop practical applications in biochemical medicine that benefit patients and the wider community. The Mechanisms in Cell Biology and Diseases Research Group has a series of research themes involving basic medical research on genetic disease, cancer, the early origins of adult health, and infection and immunity. These project areas are heavily aligned with the national research priorities of Promoting and Maintaining Good Health, A Healthy Start to Life, Aging well and Preventative Health Care. The Mechanisms in Cell Biology and Disease Research Group's primary objective is to facilitate technological advances that result in research and health outcomes that directly benefit all Australians.

• Honours, Pharmacy Honours and Doctorate of Philosophy research programs
• Physiology Essentials 100
• Molecular Pathology 300

Bachelor of Science: Flinders University of South Australia, Adelaide, South Australia. 1977.

Honours Bachelor of Science: "Studies of naturally-occurring autoimmune responses in mice." School of Biological Sciences at Flinders University, Adelaide, South Australia. 1978.

Doctorate of Philosophy: "Human B cell differentiation." Department of Clinical Immunology, School of Medicine, Flinders University of South Australia. 1982.

Affiliate Associate Professor: Faculty of Health Sciences, University of Adelaide (Affiliated through the Department of Paediatrics at the Women’s and Children’s Hospital). 2001-2012.

Professor of Molecular Medicine, Sansom Institute, School of Pharmacy and Medical Science, University of South Australia. 2006-2011.

• My research is focused on altered endosome-lysosome biogenesis in immune and mental retardation disorders. These two project areas are linked by commonality in the endosome-lysosome cell biology and related pathogenesis. For example, the current research focus on innate immune antimicrobial peptide secretion involves similar molecular mechanism to neurotransmission. Moreover, many aspects of neuropathology involve inflammation, which is becoming very evident in, for example, lysosomal storage disorder patients. The specific objectives of my research are to: Define the molecular mechanism of innate immune secretion; Devise a strategy to enable the innate immune system to eradicate H. pylori; Determine the critical link between lysosomal storage and altered neurotransmission; Develop therapeutic strategies to address mental retardation. Background: Endosomes are membrane bound compartments within eukaryotic cells and can be formed by cell surface invagination, for example during the endocytosis of macromolecules for delivery to lysosomes for degradation. Lysosomes were first described by De Duve in 1955, as acidic organelles containing an array of degradative hydrolases; a discovery that was linked with the primary dysfunction observed in lysosomal storage disorder patients. Endosomes and lysosomes are contiguous with the cell surface and therefore form a functional interface between the cell and its environment. This critical set of organelles is now recognised as having functions in macromolecular degradation, organelle turnover, cellular recycling, energy pathways, phagocytosis, pathogen killing, antigen presentation and immunity, signalling, cell membrane repair, cell division, intracellular transport, secretion and neurotransmission. Any dysfunction in endosome-lysosome organelles will therefore potentially impact on these important cellular functions. The capacity of this organelle system to respond to environmental change may act as an indicator of cellular function and disease and be altered as a result of the specific disease process. Hypotheses: Altered endosome-lysosome biogenesis is integrally involved in inflammatory and mental retardation disorders. This involvement occurs due to either a primary mutation that alters endosome-lysosome function or as part of a cellular response to another disease process. By understanding the pathophysiology of these important diseases, new diagnostic and therapeutic strategies will be identified. Current research projects include:
• 1. Investigate basic endosome-lysosome cell biology and develop innovative technologies: Generate facilities that build capacity in cell biology. Through competitive ARC LIEF grant funding and UniSA infrastructure support, basic equipment has been acquired to establish three facilities: Biophysical Characterisation (CD spectrometer; calorimeter; and Biacore), Advanced Intravital Imaging (Zeiss 710 META NLO confocal with FLIM, multiphoton, pulsed laser and spectral capacity; Vivascope with FLIM and Horiba Raman capacity), Non-invasive Analysis (2 specialised IRMS). These facilities directly support my research group as well as other SA researchers. These facilities have been achieved collaboratively with Professors John Wallace (UniA), Mike Roberts (UQ & UniSA), Clive Prestidge (UniSA), Ross Butler (UniSA) and colleagues, with a driving force and management responsibility from the Mechanisms in Cell Biology and Disease Research Group.
• 2. Targeted long lived luminescent lanthanide ion probes for live cell imaging and the study of endocytic processes (Collaboration with Dr Sally Plush). Recent advances in cell biology require accurate visualisation of biological processes in live cells at the molecular level (e.g. endocytosis, and protein trafficking). The technology to enable this visualisation of cellular processes has been partially facilitated by progress in imaging optics and instrumentation (e.g. state-of-the-art above). However, the field is limited by molecular probes with the ability to: control the method of internalisation and the targeting towards specific organelles; report on the organelle environment in the cell. Lanthanide ion probes are potentially ideal for non-invasive live cell imaging due to their small size, long emission lifetimes, minimal photobleaching and capacity for modulated emission. Hypothesis: By conjugating specific cellular targeting motifs to luminescent lanthanide ion complexes we will be able to control probe internalisation/localisation, enabling studies on the dynamic processes of sub-cellular traffic and both molecular as well as pathway interaction. Aims: 1. Generate a series of long-lived luminescent lanthanide ion probes that are conjugated to specific cellular targeting motifs. 2. Characterise the physical and photophysical properties of these molecular probes. 3. Evaluate the physiological properties of different probes in relation to both sub-compartment localisation and compartment interaction. This project will develop unique molecular probes with the distinctive photophysical properties (spectral analysis) that enable reporting on mechanisms and pathways of cell internalisation, ligand localisation and compartment interaction. For the first time, the dynamic interaction and function of specific endocytic pathways will be monitored in real time, both in vitro and in vivo.
• 3. Role of 14-3-3 proteins in regulating the innate immune response (NHMRC 631915; Collaboration with Dr Tetyana Shandala). This is the first in vivo study linking 14-3-3 proteins to innate immunity. 14-3-3 is being investigated as a key regulator for two of the most important events in an innate immune response; phagosome maturation leading to bacterial degradation; and exocytosis, which releases antimicrobial peptides (AMPs). Innate immunity is a highly conserved mechanism that eukaryotic organisms use to protect themselves against environmental challenge. Inappropriate function of innate immunity has been implicated in various high profile diseases including cancer, asthma, atherosclerosis and mental retardation disorders. Therefore, a clear understanding of the molecular mechanisms involved in generating an innate immune response, has great significance. Hypothesis: We hypothesise that the family of 14-3-3 proteins is a novel element of innate immunity, whose role is to inter-regulate two key pathways of innate immunity; phagosome maturation and anti-microbial peptide secretion. Aims: 1. Define the role of 14-3-3 in macrophage bacterial killing and phagosome maturation in hemocytes. 2. Characterise the involvement of 14-3-3 in the exocytosis of antimicrobial peptides from hemocytes. 3. Investigate the inter-relationship and cause of defective vesicle traffic in 14-3-3 deficient immune response tissues. Significance: In this project we will demonstrate that 14-3-3 is a key regulator in multiple pathways of innate immunity, elucidating roles in phagosome maturation and secretion. By identifying the molecular mechanism of 14-3-3 action, we will uncover potential points for therapeutic intervention in major inflammatory diseases where innate immunity is inappropriately regulated.
• 4. Helicobacter pylori eradication by the innate immune response (Collaboration with Prof ross Butler and Dr Glenn Borlace). H. pylori is one of the most successful human pathogens, which is estimated to have infected half of the global population. In humans, H. pylori infects the gastric mucosa causing chronic inflammation. As the principal etiological agent for gastric cancer, H. pylori is the underlying cause of 5.5% of all cancer in humans. Despite eliciting a strong innate and adaptive immune response, H. pylori persists in the host. In humans, innate immunity normally provides the critical first line of defence against pathogen challenge and macrophages are a critical component of this system. We have established a primary human macrophage model of infection and demonstrated that residual H. pylori avoid phagocytic killing in this model system for at least 48 hours (DAB 2). We have evidence that this is related to H. pylori’s capacity to alter phagosome maturation, which is the process that normally leads to bacterial degradation. By correcting phagosome maturation we will enable the immune system to clear this pathogenic bacterium and stop the onset of the chronic disease process, turning off the inflammatory cascade before it results in gastritis, ulcers and importantly gastric cancer. Hypotheses: H. pylori alters the phagosome maturation process at both the early endosome and late endosome interaction points impairing the normal process of degradation. H. pylori achieves this by producing a membrane associated protein(s) that directly interact with the phagosome vesicular machinery. Targeted disruption of the specific H. pylori protein(s) involved in this process will correct phagosome maturation and enable efficient bacterial killing and clearance. Aims:1. Define the H. pylori phagosome maturation process in primary human macrophages. 2. Determine the molecular mechanism that H. pylori uses to alter the phagosome maturation process. 3. Develop a strategy to correct the H. pylori phagosome maturation process. Significance: This project will identify the molecular mechanism that H. pylori uses to avoid macrophage killing and by targeted disruption we will devise a strategy that enables the immune system to clear this pathogenic bacterium. This will have direct significance for global health, particularly in indigenous populations where H. pylori is rife. By eradicating this bacterium there will be a direct benefit to human health by avoiding the onset of chronic disease (gastric cancer).
• 5. Role of neuronal architecture and function in lysosomal storage disorder neuropathology (Collaboration with Dr Emma Parkinson-Lawrence, Dr Kim Hemsley and Dr Damien Keating). Intense suffering and early death are common in the lysosomal storage disorders (combined prevalence of 1:5,000 live births) and more than two-thirds of patients are afflicted with progressive neurological dysfunction. The most common lysosomal storage disorder with neuropathology is mucopolysaccharidosis IIIA. There is neither a defined understanding of the basis of this neuropathology, nor an available treatment regimen. In mucopolysaccharidosis IIIA mice the accumulation of abnormal ubiquitin aggregates and multi-vesicular structures have been observed in the cell body and processes of neurons. Hypothesis: The accumulation of storage material in endosome-lysosome organelles disrupts the intracellular architecture of neurons, impacting on vesicular transport and impairing both synaptic function and neurotransmission. Aims: 1. Examine how storage impacts on the morphology and vesicular machinery of primary mucopolysaccharidosis IIIA mouse neurons. 2. Establish whether synaptic vesicle recycling and neurotransmitter release is impaired in mucopolysaccharidosis IIIA dorsal root ganglia (DRG) neurons. 3. Determine whether neuronal synaptic function is restored by enzyme replacement therapy. Significance: This project will investigate the critical link between altered endosome-lysosome function and neuropathology. By defining the impact of lysosomal storage on neuronal cell function, we will gain insight into potential sites for therapeutic intervention. The accumulation of protein aggregates and organelles in the cell body and axons of neurons has been observed in other human neurodegenerative diseases, highlighting the global significance of this research.
• 6. Development of gene delivery systems to access neuronal cells (ARC LP0883400; Collaboration with Dr Revecca Kakavanos, Prof Rob Rush, Prof Jozef Gecz). The uptake and intracellular delivery of macromolecules from the extracellular milieu is critical for the function and survival of all eukaryotic cells, and includes for example, clathrin mediated endocytosis, lipid internalisation through caveolae, macropinocytosis and phagocytosis. These pathways are directly involved in essential cellular functions that include neurotransmission, cholesterol and lipid transport, macromolecular turnover, control of cell proliferation and cell signalling. These functions involve distinct intracellular pathways and traffic to a number of specialist compartments. In this project, we will develop a successful gene transfer system for neuronal cells by comparing the uptake efficacy mediated by different endocytic mechanisms and the capacity to deliver a gene to the nucleus of target cells for expression. Our expertise in endosome-lysosome biology and the development of neuronal targeting systems by our partner company Neubody, will provide an innovative link of technologies and research experience. To evaluate gene delivery we will use Börjeson-Forssman-Lehman syndrome (BFLS) and the associated gene PHF6 as a model system. Hypotheses: Effective gene delivery can be achieved by different endocytic uptake mechanisms. Aims: 1. Determine which endocytic pathway results in efficient nuclear gene delivery and expression in neuronal cells. 2. Evaluate a new delivery system using the PHF6 gene. Significance: This project will result in an increased understanding of gene delivery and will provide a marketable product for cell biology/biotechnology, bringing economic benefit to Australia. The intracellular pathways studied, will have practical application for protein delivery systems, giving the project wider significance. A long term aim of this project is to determine whether we can efficiently deliver a gene to the brain, to enable the treatment of mental retardation disorders and other genetic diseases, which affect ~2.5% of our population.
• 7. Treatment of lysosomal storage disorder patients by drug-enhanced premature stop codon read-through (NHMRC 511321; Collaboration with Dr Makoto Kamei, Dr Maria Fuller, Prof John Hopwood). Lysosomal storage disorders are a group of over 45 different genetic disorders, each involving an enzyme deficiency of one or more lysosomal hydrolases, which results in the accumulation of undegraded substrate(s) in endosome-lysosome organelles. These disorders generally result in severe progressive pathology, which presents patients, families and health care systems with an immense burden. From studies on the molecular genetics of lysosomal storage disorders I have recognised that premature stop codons are a common type of mutation, particularly at the severe end of the clinical spectrum. In the lysosomal storage disorder mucopolysaccharidosis I, over 90% of patients have at least one premature stop codon mutation and homozygous mutations give rise to rapidly progressing pathology and mental retardation. Hypothesis: Drug-enhanced read-through of premature stop codon mutations will increase the amount of enzyme activity in lysosomal storage disorder patients and treat the biochemical disorder. Aims: 1. Define the mechanism for natural and drug-enhanced read-through of different premature stop codon mutations in vitro. 2. Evaluate stop codon read-through as a therapeutic strategy for lysosomal storage disorders. Significance: In this research project we will develop and evaluate a new therapeutic strategy for lysosomal storage disorder patients. This therapy will be directed mainly at patients from the severe end of the clinical spectrum, where treatment options are currently limited and will have direct relevance for the treatment of a wide range of other genetic disorders.

95. Parkinson-Lawrence EJ, and Brooks DA. (2007) Book Chapter: Lysosomal Biogenesis and Disease. In: Lysosomal Storage Disorders (J. Barranger ed). Springer, CRC Press, Boca Raton, Florida USA. http://www.amazon.com/Lysosomal-Storage-Disorders-John-Barranger/dp/ 0387709088/ ref=sr_1_3/105-6727723-575633?ie=UTF8&s=books&qid=1190004350&sr=8-3

96. Brooks DA, Turner C, Muller V, Hopwood JJ, Meikle PJ (2007) Book Chapter: I-cell disease. In: Lysosomal Storage Disorders (J. Barranger ed). Springer, CRC Press, Boca Raton, Florida USA. http://www.amazon.com/Lysosomal-Storage-Disorders-John-Barranger/dp/ 0387709088/ ref=sr_1_3/105-6727723-4575633?ie=UTF8&s=books&qid=1190004350&sr=8-3

97. Parkinson-Lawrence EJ, Muller VJ, Hopwood JJ and Brooks DA. (2007) N-Acetylgalactosamine-6-sulfatase protein detection in MPS IVA patient and normal control samples. Clinica Chimica ACTA, 377: 88-91.

98. Karageorgos L, Brooks DA, Harmatz P, Ketteridge D, Pollard A, Melville EL, Parkinson-Lawrence E, Clements PR, and Hopwood JJ. (2007) Mutational analysis of mucopolysaccharidosis type VI patients undergoing a phase II trial of enzyme replacement therapy. Molecular Genetics and Metabolism, 90: 164-170.

99. Brooks DA. (2007) Getting into the fold. Nature Chemical Biology, 3: 84-85.

100. Shoubridge C, Cloosterman D, Parkinson-Lawrence EJ, Brooks DA and Gecz J. (2007) Molecular pathology of mutations in the ARX homeobox gene. Genomics, 90: 59-71.

101. Karageorgos L, Brooks DA, Pollard A, Melville EL, Hein LK, Clements PR, Ketteridge D, Swiedler SJ, Beck M, Giugliani R, Harmatz P, Wraith JE, Guffon N, Sá Miranda MC, Teles EL, and Hopwood JJ. (2007) Mutational analysis of 105 mucopolysaccharidosis type VI patients. Human Mutation, 28: 897-903.

102. Tarpey PS, Raymond FL, Nguyen LS, Rodriguez J, Hackett A, Shoubridge C, Vandeleur L, Smith R, Edkins S, Stevens C, O’Meara S, Tofts C, Barthorpe S, Buck G, Cole J, Halliday K, Hills K, Jones D, Mironenko T, Perry J, Varian J, West S, Widaa S, Teague J, Dicks E, Butler A, Menzies A, Richardson D, Jenkinson A, Shepherd R, Raine K, Moon J, Luo Y, Parnau J, Baht SS, Gardner A, Corbett M, Brooks DA, Thomas P, Parkinson-Lawrence EJ, Porteous M, Sanderson T, Pearson P, Simensen RJ, Skinner C, Hoganson G, Superneau D, Easton DF, Wooster R, Bobrow M, Turner G, Partington M, Stevenson RE, Futreal PA, Schwartz CE, Srivastava AK, Stratton MR and Gécz J. (2007) Loss of function mutations in UPF3B, a member of the nonsense-mediated mRNA decay surveillance complex, cause mental retardation. Nature Genetics, 39: 1127-1133.

103. Mardones GA, Burgos PV, Brooks DA, Parkinson-Lawrence EJ, Mattera R and Bonifacino JS. (2007) The TGN accessory protein p56 cooperates with the GGA adaptors in the biosynthetic sorting of cathepsin D to lysosomes. Molecular Biology of the Cell, 18: 3486-3501.

104. Borlace GN, Butler RN, and Brooks DA. (2008) Monocyte and macrophage killing of Helicobacter pylori: relationship to bacterial pathogenicity factors. Helicobacter, 13:380-387.

105. Brooks DA and Fuller M. (2009) Lysosomal disorders. Wiley Encyclopedia of Chemical Biology. 4: 1-11.

106. Brooks DA. (2009) The endosomal network. International Journal of Clinical Pharmacology and Therapeutics, 47: Suppl 1: S9-S17.

107. Morrison JL, Wang KCW, Brooks DA, and Botting KJ. (2009) Fetal heart growth: Insulin-like growth factor 1 and sex. Expert Reviews in Obstetrics and Gynecology, 4: 1-5.

108. Keep SJ, Borlace GN, Butler RN, and Brooks DA. (2010) Role of immune serum in the killing of Helicobacter pylori by macrophages. Helicobacter, 15: 177-183.

109. Parkinson-Lawrence EJ, Shandala T, Prodoehl M, Plew R, Borlace GN and Brooks DA. (2010) Lysosomal storage disease – revealing lysosomal function and physiology. Physiology, 25: 102-115.

110. Shandala T, Parkinson-Lawrence EJ and Brooks DA. (2010) Protein cotranslational and posttranslational modification in organelles. Encyclopedia of Life Sciences. Copyright © 2010 John Wiley & Sons, Ltd.

111. Jones HF, Burt E, Dowling K, Davidson G, Brooks DA, and Butler RN. (2010) The effect of age on fructose malabsorption in children presenting with gastrointestinal symptoms. Journal of Pediatric Gastroenterology & Nutrition. In press September 2010.

112. Jones HF, Davidson G, Brooks DA, and Butler RN. (2010)Small bowel bacterial overgrowth in children with suspected carbohydrate malabsorption – an under-diagnosed disorder? Journal of Pediatric Gastroenterology & Nutrition, In press 6th December 2010.

113. Jones HF, Butler RN, and Brooks DA. (2010) Fructose transport and malabsorption in humans. American Journal of Physiology - Gastrointestinal and Liver Physiology, In press 6th December 2010.

114. Borlace GN, Jones HF, Keep SJ, Butler RN, and Brooks DA. (2011) Helicobacter pylori phagosome maturation in primary human macrophages. Gut Pathogens, In press 28th February 2011.

115. Shandala T, Woodcock J M, Ng Y, Biggs L, Skoulakis EMC, Brooks DA, and Lopez AF. (2011) Drosophila 14-3-3å has a critical role in anti-microbial peptide secretion and innate immunity. Journal of Cell Science, In press 15th March 2011.

I am able to provide media comment in the following areas of expertise:

Discipline: Biochemistry, Cell Biology

• Immunochemistry & Cell Biology
• Endosomes & Lysosomes
• Cancer cell biology
• Innate immunity
• Intracellular neurobiology