The role of BCL3 in high risk plasma cell myeloma

Multiple myeloma (MM), a chronic and incurable plasma cell malignancy, accounts for approximately 2% of all cancer deaths worldwide. The median age of disease onset is approximately 65 years, with an average life expectancy of approximately 7-8 years from the time of diagnosis. However, about 20% of patients at diagnosis have a median survival of <2 years. These individuals are referred to as “ultra high-risk” MM patients. While most patients with MM respond to therapy, “ultra high-risk” MM patients respond poorly to therapeutic regimens, including the novel agents, thus highlighting the need to understand the underlying molecular mechanisms that of this disease subgroup. High BCL3 expression, an NF-κB co-activator, has been purported to be a feature of proliferative MM; often associated with a poor prognosis.

Previous observations made by our laboratory using an independent patient cohort recapitulate these findings, demonstrating that patients with elevated BCL3 expression have inferior progression free survival (PFS) and overall survival (OS). Despite this striking clinical observation, the biological consequences of BCL3 expression in MM remain poorly understood. For this reason, we are interested in (i) determining whether high levels of Bcl-3 could be used as a biomarker to identify ultra high-risk MM; (ii) establishing the mechanism of Bcl-3 upregulation in MM, and (iii) understanding the functional consequence of Bcl-3 upregulation in MM. Research of this nature is feasible in human MM cell lines, as we can alter levels of BCL3 across various cell lines and observe the impact this has on MM cell proliferation and survival. We will utilise the CRISPR/Cas9 methodology to edit the BCL3 gene in myeloma cell lines in an inducible manner. To complement these experiments, we will also overexpress Bcl-3 in cell lines that express low levels of Bcl-3 and assess cell growth and proliferation.

Collaborators

Dr Pasquale Fedele
Dr Michael Low
Dr Raffi Gugasyan

Understanding the Biology of the Myelodysplastic Syndromes

Myelodysplastic syndromes (MDS) are a heterogenous group of clonal haematopoietic stem cell malignancies causing low blood counts with a propensity to transform to acute leukaemia. This blood cancer occurs primarily in the elderly, and the incidence of this disease will continue to increase with our ageing population. The only potentially curative therapy for MDS is an allogeneic haematopoietic stem cell transplant, which is not a feasible option in the majority of these elderly patients with other precluding medical comorbidities. The mainstay of treatment for the patients with lower risk disease, who make up almost 75% of MDS patients, is supportive management with blood transfusions which carry a risk of iron overload.

Consistent with a number of observational studies, our lab has also found haematopoietic improvement in patients with low risk disease treated for iron overload with the iron chelator Deferasirox (DFX).

We have preliminary data from our laboratory demonstrating that iron chelation blocked cell growth and proliferation of myeloid cell lines with mitochondrial dysfunction and apoptosis of malignant clones. We hypothesise that this iron chelator has a role in autophagy, a cellular recycling process that contributes to the pathogenesis of this disease.

We have looked at the effect of iron modulation on autophagy in myeloid cell lines. Utilising the CRISPR/Cas9 genome editing to knockout essential autophagy genes in various cell lines, we will be looking at the effect of this cellular process in the propagation of the malignant clone as well as erythroid differentiation, as MDS are characterised by ineffective erythropoiesis.  We are also looking at iron modulation in a mouse model of MDS and its effect on halting progression of disease.

With increasing knowledge of immune dysfunction in MDS and its impact on transformation of disease from low risk to high risk, we are also looking at exploring immunomodulatory mechanisms as potential disease modifying agents.

Collaborators

Dr Chris Slape
A/Professor Paul Ekert
A/Professor Ron Firestein
Prof Stephen Opat

Title: Role of TIMPs in Ovarian Cancer

Eighty per cent of ovarian cancer patients die within the first five years of diagnosis due to recurrence associated with chemoresistance.  The proposed study will investigate the role of tissue inhibitors of matrixmetalloproteinases [TIMPs] which regulate extracellular matrix [ECM] remodelling important for metastasis and affect the survival of ovarian tumour cells before and after chemotherapy treatment.

In normal tissues the main role of TIMPs is to inhibit metalloproteinases (such as MMPs, ADAMs and ADAMTS).  However in ovarian cancer, before or after chemotherapy, the balance between TIMPs and matrixmetalloproteinases is changed when compared to normal tissues.

We hypothesize that alterations in TIMP signaling are associated with ovarian tumour progression and promote chemoresistance and recurrence in ovarian cancer.  The aim of this project is to do a comprehensive analysis of genes and proteins associated with TIMP signalling in primary tumours, chemonaive and recurrent tumours and tumour cells present in ascites fluid obtained from patients with ovarian cancer.  This project will also genetically manipulate ovarian cancer cell lines and use in vitro functional assays and mouse xenograft models to better understand the role of TIMP signalling in ovarian cancer progression.

Collaborators: 

Dr Harry Georgiou (University of Melbourne)

Transfusion-dependent thalassemia and bone disease

Thalassemia is a disorder of haemoglobin synthesis due to mutations in the globin chains (α or β). In its more severe form it requires treatment with chronic transfusion therapy which leads to iron overload with cardiac, liver, endocrine and bone complications. Monash Health is the state referral centre for the management of transfusion-dependent thalassemia. Our recent work in conjunction with Professor Don Bowden (Head of Thalassemia Services) has described for the first time, a case of reversible osteomalacia secondary to Fanconi’s syndrome in the setting of an iron chelator, with ongoing studies examining mechanisms of bone loss in this population. We have also recently reported a high prevalence of kidney stones in transfusion-dependent thalassaemia and an association between reduced bone density, kidney stones and fractures.

We are exploring the kidney / bone axis as it is now becoming apparent that a number of clinical conditions impact these organs. Whilst these organs have been considered in isolation in the past, the interdependence of both organs is paramount in the skeletal integrity.

Collaborators:

Professor Don Bowden
Professor Peter Kerr

Using a three-dimensional in vitro mouse ovarian culture model to create an artificial ovary

test_tubes1Many young female cancer patients undergo chemo- or radiation therapy, which introduces a risk of infertility. The in vitro growth and maturation of the ovarian follicle, which is the basic functional unit of the ovary, provides the potential to preserve fertility by growing follicles in vitro and transplanting them back into the patient. Growth and maturation of secondary follicles into large pre-ovulatory follicles, is essential to gaining insights into human folliculogenesis. Thus, the three-dimensional alginate in vitro follicle culture system is an excellent model to study the role of growth factors, metabolites and physical environment of the ovary.

Fertility preservation in female cancer patients

ovaryIrreversible damage to the ovary is a devastating side effect of many anti-cancer treatments, often leaving cancer survivors unable to have their own children and facing premature menopause. We are investigating new strategies to protect the ovary from radiation and chemotherapy induced damage in order to preserve fertility and ovarian function in these patients.

Anti-cancer treatments, including both radiation and chemotherapy, are associated with significant immediate and long-term health problems for cancer survivors, including serious adverse effects on reproductive function and ovarian hormone production. In particular, these treatments can damage the DNA of eggs and induce their death, leading to premature ovarian failure and infertility. Currently, no options exist to protect the ovary from damage and preserve fertility of young women being treated for cancer.

Our team is investigating new strategies to protect the ovary and preserve fertility following radiation and chemotherapy through the inhibition of egg death. Using gene targeted mouse models, we have found that elimination of the potent cell killers PUMA and NOXA, protects eggs from radiation-induced death. We have also shown that the surviving eggs can repair their damaged DNA and are of sufficient quality to produce healthy offspring.

These studies provide strong support for the concept that fertility may be safely and effectively preserved during anti-cancer treatment by inhibiting genes required for egg death, such as PUMA and NOXA. We are now working with our collaborators from the Walter & Eliza Hall Institute and the Royal Women’s Hospital to confirm our findings in the human ovary and to develop novel, clinically relevant, fertoprotective adjuvant therapies.

Collaborators:

  • Monash University, Melbourne
  • Walter & Eliza Hall Institute, Melbourne
  • Melbourne IVF
  • University of Edinburgh

Egg supply, the fertile lifespan, and age at menopause

ovaryWomen are born with a limited supply of eggs (oocytes) in their ovaries and are unable to make new eggs after birth. Because of this, the number and health of eggs established within the ovary early in life influence the length of time for which a female will be fertile, her age at menopause, and the health of her offspring. We are investigating the mechanisms that control egg supply and health during ovarian development and throughout reproductive life.

A striking characteristic of normal ovarian development is the extensive, but unexplained, death of the embryonic precursors of eggs (germ cells), leaving a relatively small number of eggs stored in the ovary at birth to sustain fertility and ovarian function throughout life. Why are such a large number of germ cells generated during embryonic development then destroyed? Are these destroyed germ cells eliminated because they are of low quality? What are the genes and proteins that regulate germ cell death?

Answering these questions and understanding the mechanisms that determine how many eggs are established and maintained in the ovary is essential, as abnormal regulation of death pathways leading to reduced egg number may compromise female fertility and  result in pre-mature menopause. Our work is therefore highly relevant to female fertility and health, as premature menopause not only reduces a woman’s chance of having children, but also puts her at early risk of post-menopausal problems such as osteoporosis and heart disease.

Our laboratory, together with colleagues from the Walter & Eliza Hall Institute and the University of Edinburgh, is investigating the genes and proteins involved in determining whether a germ cell will live or die. We have discovered that a family of cell death proteins, known as BH3-only proteins, play critical roles in the initiation of germ cell death and act to limit the size of the germ cell pool before birth. We are now determining if it is possible to prevent germ cell death and increase the number of eggs available in adult life to prolong fertility and delay menopause by inhibiting or eliminating BH3-only proteins.

Collaborators:

  • Monash University, Melbourne
  • Walter & Eliza Hall Institute, Melbourne

Fetal growth restriction

Using pre-clinical models of fetal growth restriction (FGR), the group has sought to better understand fetal adaptations to impaired placentation and thereby develop better diagnostics and therapies. Dr Suzie Miller, head of the Neurodevelopment and Neuroprotection group, is driving much of the fundamental (experimental) work in this area. Insights gained from the experimental studies have informed the design of clinical trials of melatonin as a neuro- and cardio-protectant, reducing the risks of brain injury and cardiovascular impairment. Follow-up studies of the children are being led by Professor Rosemary Horne and Associate Professor Michael Fahey. A world first trial was recently completed using melatonin in pregnant women to prevent cerebral palsy.

Collaborators:

Professor Rosemary Horne – NHMRC Senior Research Fellow, Department of Paediatrics
Associate Professor Michael Fahey – Paedatric Neurologist, Department of Paediatrics
Dr Stephanie Yiallourou – Senior Postdoctoral Research Fellow, Department of Paediatrics

Preeclampsia

Preeclampsia remains one of the leading causes of maternal and perinatal mortality and morbidity worldwide. It is now the major cause of iatrogenic preterm birth in Australia. There have been no therapeutic advances in the management of preeclampsia for almost 50 years. The Group continues to explore fundamental mechanisms of maternal endothelial injury in preeclampsia with a view to the development of more effective therapies. The research involves in vitro studies, small animal models of preeclampsia, and clinical trials. A world first clinical trial of melatonin as an adjuvant therapy for preeclampsia is underway. The Group is also leading work on the role(s) of activin and other TGF family members in the pathogenesis of preeclampsia. Dr Bryan Leaw recently joined the group to establish a panel of pre-clinical models for therapeutic testing.

Collaborators:

Professor Milton Hearn– Director, Centre for Bioprocess Technology, Department of Biochemistry and Molecular Biology
Associate Professor Grant Drummond – NHMRC Senior Research Fellow, Department of Pharmacology, Monash University
Associate Professor Chris Sobey – Department of Pharmacology, Monash University

Amnion cell therapies

As the world’s leading research group on amnion epithelial cells in regenerative medicine, the Maternal and Perinatal Research Group has led the application of amnion cells as a therapy for preterm lung disease. The team will commence a world first clinical trial in 2014. The amnion cell research is a large component of the Cell Therapy and Regenerative Medicine Theme within the Centre, reflected by the multiple collaborations between groups within that theme including Dr Rebecca Lim’s Amnion Cell Biology group and Professor Graham Jenkin’s Cell Therapy group).The Group also collaborates closely with Associate Professor Tim Moss’ Fetal and Neonatal Health group on large animal models of lung injury and the application of amnion cells. The Group is now extending the research application of amnion cells to other conditions of the extreme preterm infant including necrotizing enterocolitis, using animal models to direct the design of future clinical trials.

Collaborators:

Professor Bill Sievert – Clinician Scientist, Director of Gastrointestinal and Liver Unit, Monash Health and Monash University
Associate Professor Tim Moss – NHMRC Senior Research Fellow, Department of Obstetrics and Gynaecology, Monash University
Associate Professor Chris Sobey – NHMRC Senior Research Fellow, Department of Pharmacology, Monash University
Professor Claude Bernard – Monash Immunology and Stem Cell Labs, Monash University
Professor Daniel Chambers – Thoracic and Transplant Physician, School of Medicine, University of Queensland