Kunjin Replicons For Gene Therapy And Protein Manufacture
Funder
National Health and Medical Research Council
Funding Amount
$310,000.00
Summary
This grant seeks to provide proof of concept (PoC) for the use of the Kunjin replicon technology for gene therapy and protein production. (A) Protein production. Two Kunjin replicon constructs expressing green fluorescent protein (GFP) and secreted alkaline phosphatase (SEAP) are to be constructed and protein production monitored using FACS and SEAP bioactivity reporter kit (Roche), respectively. Protein production and biological activity of the proteins will be monitored in transient transfecti ....This grant seeks to provide proof of concept (PoC) for the use of the Kunjin replicon technology for gene therapy and protein production. (A) Protein production. Two Kunjin replicon constructs expressing green fluorescent protein (GFP) and secreted alkaline phosphatase (SEAP) are to be constructed and protein production monitored using FACS and SEAP bioactivity reporter kit (Roche), respectively. Protein production and biological activity of the proteins will be monitored in transient transfections and over an extended time period. Several cell lines, culture conditions and Kunjin replicon vector modifications will be tested. Arrangements have also been made to send the constructs to Roche, GSK, Eli Lilly, and Exelixis for side by side comparisons of this system with existing proprietary protein production echnologies. (B) Gene therapy. Two PoC gene therapy systems are proposed to be used for evaluation of Kunjin replicon vectors. (i) Tumours expressing granulocyte macrophage colony stimulating factor (GMCSF) by transfection cause the generation of anti-tumour CD8 T cells and subsequent tumour rejection. Current approaches include adoptive transfer of adeno-GM-CSF transfected tumour cells, a costly and laborious process resulting in only transient expression (Can. Imm. Immunother 2001 50:373). We intend to inject Kunjin replicon virus like particles into growing s.c. B16 melanomas and expect to see a high infection rate, a sustained high-level expression of GMCSF, and rejection of the tumour. In contrast to Kunjin, nearly all humans have antibody responses to adenovirus, and very high titres of adenovirus are required to obtain high infection and GM-CSF expression. Both factors limit adenovirus use in vivo. (ii) Transplant rejection can be inhibited by expression in the graft of CTLA4-Fc a reagent that blocks T cell co-stimulation enhancing allo-graft acceptance (Transplantation 2000 69:1806). High-level expression for over 100 days is expected to correlate with optimal graft acceptance. Our ability to use Kunjin to express beta galactosidase for several months in vivo without inflammation illustrates the potential for this approach (CIB ref 15). Initially we intend to use P815 cells injected i.p. into C57BL-6, where they are usually rejected within a few days. In contrast, P815 cells with Kunjin replicon-mediated CTLA4-Fc expression should survive for an extended period. Graft survival is easily monitored using FACS and anti-H-2d antibodies.Read moreRead less
Performance And Safety Testing Of The BioQ Cardiac Assist System In A Chronic Ovine Heart Failure Animal Model
Funder
National Health and Medical Research Council
Funding Amount
$142,800.00
Summary
This proposal will test a novel cardiac assist system in safety and performance studies using a chronic sheep heart failure model. This device has been tested in cardiovascular simulators and in an acute animal model showing attractive proof-of-concept data. Specifically, the device increased left coronary artery blood flow and reduced aortic pulse and mean pressures using our novel self-powered fully implantable stand alone device, a potential therapy treatment for heart failure.
The Effect Of Stress/strain And Fatigue Fracture Sites On Durability Of Modular Aortic Endografts And Arterial Walls
Funder
National Health and Medical Research Council
Funding Amount
$300,919.00
Summary
Aneurysmal disease is an age related phenomenon. The mean life expectancy of western populations has doubled in 100 years because of the reduction in deaths from preventable and treatable diseases, and prolongation of life with chronic and incurable diseases. The older community (>65 years) continues to be active and productive contrary to prior predictions. Aneurysmal disease has emerged as a result of the changing pattern of diseases in the community. As with many other diseases, prophylaxi ....Aneurysmal disease is an age related phenomenon. The mean life expectancy of western populations has doubled in 100 years because of the reduction in deaths from preventable and treatable diseases, and prolongation of life with chronic and incurable diseases. The older community (>65 years) continues to be active and productive contrary to prior predictions. Aneurysmal disease has emerged as a result of the changing pattern of diseases in the community. As with many other diseases, prophylaxis against aneurysmal disease is the most effective approach since 80% of those that rupture will result in death. Endoluminal grafting provides a much less invasive procedure and provides an attractive and elegant alternative to open surgery. The danger is that structural strengths will be compromised with failures due to lack of strength and inadequate device durability. The most practical endografts are those that are built up from modules but vulnerable sites affected by fatiguing and disruptive forces are being identified, in particular where a tube divides into two outflow channels – the bifurcation - and joins. To improve and protect the current device, and enable sound engineering for future devices, we need to know the nature, magnitude and location of these forces. The existing collaboration with medical specialist, Cook Aust., the Advanced Fluid Dynamics Laboratory (CSIRO, Melb.) and University of WA combines a multidisciplinary team working towards the design and durability of the next generation of endoluminal devices and percutaneous insertions. While Australia is at the forefront of development, its current place in the market can only be maintained by further product development. Optimising and improving endoluminal grafts will reduce the suffering of major surgery while prolonging a higher quality of life with a much less invasive procedure whose long-term effectiveness, and thereby acceptance is dependent upon reliable durability.Read moreRead less
Mechanical Mobility Of The Thorax For Continuous Determination Of Lung Gas Volume
Funder
National Health and Medical Research Council
Funding Amount
$165,000.00
Summary
Percussion is a valuable clinical method for physical examination of parts of the body. A sharp tap (impulsive force) is applied to the body wall and the sound radiated in response is observed. This sound may be dull (over liver) or stony dull (pleural effusion), or resonant (over normal lung) or hyper-resonant (over bowel). While the variation in radiated sound is not fully understood, it is apparent that the presence of gas, which is highly compliant, increases mobility of the overlying tissue ....Percussion is a valuable clinical method for physical examination of parts of the body. A sharp tap (impulsive force) is applied to the body wall and the sound radiated in response is observed. This sound may be dull (over liver) or stony dull (pleural effusion), or resonant (over normal lung) or hyper-resonant (over bowel). While the variation in radiated sound is not fully understood, it is apparent that the presence of gas, which is highly compliant, increases mobility of the overlying tissue and allows it to resonate; where the sub-tissue is largely fluid, tissue mobility is low and the percussive sound is dull. Percussion is useful for examining the adult chest and lung, but cannot for example be applied in infant intensive care as only limited impulsive force can be used, and the adult finger, which is both a coupling device and sounding board, is too large. As well, percussion requires skill and quiet conditions. Accordingly, we developed a device to measure mobility of the chest and other tissues in real time. The VibroPulse applies a known low-level force to the body surface and records the resultant velocity induced in the surface. The force is generated by a vibrating mass set in motion by an electromagnetic motor driven by pseudo-random noise. Tissue mobility, defined as velocity-force, is derived simultaneously across the frequency range, providing an easily interpreted quantitative output unaffected by ambient noise. This proposal has two aims we can achieve in 1 year: (1) to continue evaluating VibroPulse sensitivity to tissue composition, using symmetrical percussive sites on the human chest and abdomen that are dull on one side and resonant on the other, and the chest of anaesthetised animals with experimentally induced pneumothorax and lung collapse, two life-threatening conditions for which percussion is a key diagnostic method, and (2) to engineer a small device from our bulky prototype that is suitable for clinical use, in infants and adults.Read moreRead less
Development And Evaluation Of Novel Fetal Haemoglobin Inducers For The Therapy Of Beta-thalassaemia
Funder
National Health and Medical Research Council
Funding Amount
$288,899.00
Summary
The most important haemoglobinopathies from the clinical point of view are the beta-thalassaemias, sickle cell disease (SCD), HbE disease and the interactions between them. These beta-haemoglobinopathies are the result of mutations in the beta-globin gene, causing beta-globin chain synthesis that is abnormal, low or absent leading to life-threatening severe anaemia, and blood transfusion-dependency for life. An alternative approach to the therapy of beta-thalassemia is to reactivate fetal haemog ....The most important haemoglobinopathies from the clinical point of view are the beta-thalassaemias, sickle cell disease (SCD), HbE disease and the interactions between them. These beta-haemoglobinopathies are the result of mutations in the beta-globin gene, causing beta-globin chain synthesis that is abnormal, low or absent leading to life-threatening severe anaemia, and blood transfusion-dependency for life. An alternative approach to the therapy of beta-thalassemia is to reactivate fetal haemoglobin (HbF) synthesis. Some chemical agents have been identified to induce HbF and significantly reduce the need for blood transfusion in some thalassaemia patients, while in SCD patients it can ameliorate the clinical symptoms. Despite a number of clinical trials investigating the potential of HbF-inducing agents, many of these drugs have low efficacy, specificity, and cytotoxicity. There is therefore an urgent need to identify novel pharmacological agents with greater efficacy and reduced toxicity. Without a clear understanding of the underlying mechanism(s) involved in the induction of HbF, it is virtually impossible to focus on any molecular target. A promising approach is the use of chemical libraries in a high-throughput (HTP) screening to identify positive regulators of gene products. Our research group created an assay that has allowed us for the first time to perform a side-by-side comparison of several previously described fetal hemoglobin inducers including 2000 existing pharmaceuticals used by patients unrelated to thalassaemia. The screen identified a distinct group of compounds that induced the gamma-globin promoter in primary and secondary screens. The identification of novel inducers of HbF warrants further investigation as alternative therapies for beta-thalassemia. This project will evaluate novel inducers of HbF in our thalassaemia mouse model and provide early 'proof-of-concept' and enable the initiation of preclinical and clinical studies.Read moreRead less
Development Of Non-surgical Approach To Treating Tricuspid Regurgitation
Funder
National Health and Medical Research Council
Funding Amount
$266,427.00
Summary
Heart failure is a common problem in which the heart enlarges and contracts poorly. In association with enlargement of the heart, the heart valves also begin to fail causing further worsening of quality and length of life. Failure of the tricuspid valve occurs in upto 87% of patients with heart failure and presently the only treatment option is high risk heart surgery. We are developing a way of dealing with tricuspid valve failure that does not require cardiac surgery.
Novel Nanoparticle Composites For Molecular Probes In Diagnostic Imaging
Funder
National Health and Medical Research Council
Funding Amount
$170,716.00
Summary
Isotope labelled protein probes, eg. antibodies, are a valuable imaging tool in investigating patient disease. Their biological specificity is their great strength, however, detection sensitivity often limits their use. A novel nanoparticle developed at ANU can increase this signal by a million-fold in comparison with conventional methods of labelling. This approach suits a range of probes and will accommodate many of the isotopes already used in patient diagnostics and therapy.
Innovations For Better Cardiovascular Prevention In Primary Care
Funder
National Health and Medical Research Council
Funding Amount
$294,285.00
Summary
The goal of this project is to take a new concept for cardiovascular disease management through the proof-of-concept stage and deliver a novel model for cardiovascular disease management. When proven, the application of this new process for service delivery will provide a low-cost way of closing the gap between guideline recommendations and current management of vascular disease in Australian primary care settings.
Development Of A Vaccine For Genital Chlamydial Infection
Funder
National Health and Medical Research Council
Funding Amount
$207,551.00
Summary
Genital Chlamydia infections are the most common sexually transmitted infection in Australia with annual health costs of 90-160 million dollars. Infection rates in 15-29 olds are increasing at 15-20% per year. Antibiotics are currently the treatment of choice, however antibiotic resistance is increasing and most infections are asymptomatic and not treated in the absence of screening programs. This project aims to develop a genital Chlamydia vaccine using a combination of novel antigens.
Minimal Structural Strength Required In Endoluminal Devices For Aortic Aneurysms: Mechanisms Of Rupture When They Fail.
Funder
National Health and Medical Research Council
Funding Amount
$102,794.00
Summary
Aneurysmal disease is an age related phenomenon. The mean life expectancy of western populations has doubled in the last 100 years mainly because of the reduction in deaths from occlusive cardiovascular disease. Aneurysmal disease has emerged as a result of the changing pattern of diseases in the community. The impact of this work would be to improve minimally invasive, effective prophylaxis against aneurysm of the abdominal aorta since 80% of those that rupture will result in death. Endoluminal ....Aneurysmal disease is an age related phenomenon. The mean life expectancy of western populations has doubled in the last 100 years mainly because of the reduction in deaths from occlusive cardiovascular disease. Aneurysmal disease has emerged as a result of the changing pattern of diseases in the community. The impact of this work would be to improve minimally invasive, effective prophylaxis against aneurysm of the abdominal aorta since 80% of those that rupture will result in death. Endoluminal grafting provides a much less invasive procedure and provides an attractive and elegant alternative to open surgery. The general drive in this field is towards miniaturisation. The danger is that structural strengths will be compromised with early failures due to lack of strength; and late failures due to inadequate device durability. This application proposes the development of a collaborative venture with the Department of Vascular Surgery, Royal Perth Hospital, Cook R and D (WA), the Advanced Fluid Dynamics Laboratory (CSIRO, Melbourne) and Centre for Health Services Research (University of WA) to investigate the dynamic flow forces involved in aneurysms of the abdominal aorta. It combines together expertise and facilities to examine the forces acting upon endoluminal stent grafts in pulsatile high pressure environments, the impact on the design, patency, effectiveness and longer-term outcomes. The multidisciplinary team provides a comprehensive approach for this developing area of high technology surgery throughout Australia and internationally. The benefits of this study will directly influence the design and durability of the next generation of endoluminal devices and percutaneous insertions. While Australia is at the forefront of development, its current place in the market can only be maintained by further product development.Read moreRead less