Replacement of animal models for tumour biology with a multifunctional microfluidic-based approach
Lead Research Organisation:
University of Hull
Department Name: Postgraduate Medical Institute
Abstract
The aim of the project is to design and optimise a novel microfluidic device in which small biopsies of human tumour tissue can be kept alive, treated with drugs and the effects tested downstream by appropriate analysis techniques. This human system will be able to replace the generalised testing of new drugs and combinations of drugs by the pharmaceutical industry, on animal models that only poorly replicate the physiology and metabolism of human beings. In addition it will be possible to test biopsies of both normal and diseased pieces of any tissue type so that both specific therapeutic responses, as well as non-specific side-effects can easily be assessed.
There is growing evidence that patients with many cancers, particularly ovarian and pancreas suffer with an increased occurrence of blood clotting events; these clots can be lethal. There is also evidence that when patients undergo chemotherapy and the tumour cells are destroyed the release of cell debris further increases the risk of clotting. In the project we propose to design a device that allows analysis of these clotting pathways, using a human ovarian tumour model. Three analysis modules will be integrated into a device on which a sample of tissue is incubated, allowing different types of biological response to be monitored, including changes in the expressed genes, cell surface components and secreted products. These distinct types of analysis module demonstrate the widespread applicability and potential of this new platform technology. Furthermore, ovarian cancer has a relatively high mortality with survival rates not improving substantially over the past two decades.
Not only will the new technology replace a large number of conventional animal tests, of limited proven value, but the capability of undertaking multiple tests on a tumour sample, that is an extremely close representation of the mass from which it came, after the tissue has been subjected to a drug or other test condition, means that a large amount of information can be generated on any individual tumour. Such data will be extremely useful for the clinician when obtained at the time of diagnosis. The intended treatment strategy can be tailored to the patient, and where no treatment appears to give any therapeutic effect, this can be discussed with the patient opening the opportunity for palliative care to be the preferred choice, whilst maintaining the best quality of life.
There is growing evidence that patients with many cancers, particularly ovarian and pancreas suffer with an increased occurrence of blood clotting events; these clots can be lethal. There is also evidence that when patients undergo chemotherapy and the tumour cells are destroyed the release of cell debris further increases the risk of clotting. In the project we propose to design a device that allows analysis of these clotting pathways, using a human ovarian tumour model. Three analysis modules will be integrated into a device on which a sample of tissue is incubated, allowing different types of biological response to be monitored, including changes in the expressed genes, cell surface components and secreted products. These distinct types of analysis module demonstrate the widespread applicability and potential of this new platform technology. Furthermore, ovarian cancer has a relatively high mortality with survival rates not improving substantially over the past two decades.
Not only will the new technology replace a large number of conventional animal tests, of limited proven value, but the capability of undertaking multiple tests on a tumour sample, that is an extremely close representation of the mass from which it came, after the tissue has been subjected to a drug or other test condition, means that a large amount of information can be generated on any individual tumour. Such data will be extremely useful for the clinician when obtained at the time of diagnosis. The intended treatment strategy can be tailored to the patient, and where no treatment appears to give any therapeutic effect, this can be discussed with the patient opening the opportunity for palliative care to be the preferred choice, whilst maintaining the best quality of life.
Technical Summary
The aim of the project is to design and optimise a microfluidic device in which small biopsies of human tissue (malignant and normal) can be maintained in a biomimetic-like environment. It is proposed that these devices will be used both by pharmaceutical companies to reduce and replace drug screening and by clinicians as tools for personalising a therapeutic strategy. It is planned to investigate the pro-coagulant changes that occur in many cancers as the process is a major clinical problem and provides an ideal setting to demonstrate the widespread applicability and potential of the microfluidics platform.
To investigate the reasons why a venous thromboembolism is commonly associated with tumours, tissue biopsies (2-3m3) will be maintained in a small chamber on the chip (approximately 20 ?Yl in volume) for up to 8 days with media, or media plus drugs at clinically-relevant doses, perfusing the tissue. A major benefit of the microfluidic approach is that not only can drug combinations be tested, but also the most effective sequence of administering drugs can be rationally determined. This type of analysis is not possible in animal models as the one end point is almost always death. Additionally, normal tissue will be tested in the devices allowing side effects such as non-specific toxicity to be identified.
It is proposed to establish three distinct analysis modules measuring different facets of the clotting process: coagulation, using viscosity and direct fluorescence imaging; MP release, by dual colour flow cytometry; and gene transcription using hybridisation and fluorescence detection. The modules can be used in parallel due to a switch mechanism on the chip and on multiple occasions, thus a response can be monitored over time. These integrated devices will provide data on the responses that have far greater relevance to the in vivo setting than any animal model. A key aim of the proposal is to demonstrate the robustness of the microfluidic device and compatibility of the data generated on-chip with conventional cell assays.
Having established the operating parameters of the device it is proposed to test a cohort of ovarian tumour samples using a standardised chemotherapy regimen (paclitaxel plus carboplatin) alone, and in combination with a currently used VTE treatment. The results of this microfluidic trial will be correlated with the clinical outcome. Finally, a small group of tumours will be used to establish the optimal drug concentrations and dosing sequence for each tumour, i.e. a personalised treatment.
To investigate the reasons why a venous thromboembolism is commonly associated with tumours, tissue biopsies (2-3m3) will be maintained in a small chamber on the chip (approximately 20 ?Yl in volume) for up to 8 days with media, or media plus drugs at clinically-relevant doses, perfusing the tissue. A major benefit of the microfluidic approach is that not only can drug combinations be tested, but also the most effective sequence of administering drugs can be rationally determined. This type of analysis is not possible in animal models as the one end point is almost always death. Additionally, normal tissue will be tested in the devices allowing side effects such as non-specific toxicity to be identified.
It is proposed to establish three distinct analysis modules measuring different facets of the clotting process: coagulation, using viscosity and direct fluorescence imaging; MP release, by dual colour flow cytometry; and gene transcription using hybridisation and fluorescence detection. The modules can be used in parallel due to a switch mechanism on the chip and on multiple occasions, thus a response can be monitored over time. These integrated devices will provide data on the responses that have far greater relevance to the in vivo setting than any animal model. A key aim of the proposal is to demonstrate the robustness of the microfluidic device and compatibility of the data generated on-chip with conventional cell assays.
Having established the operating parameters of the device it is proposed to test a cohort of ovarian tumour samples using a standardised chemotherapy regimen (paclitaxel plus carboplatin) alone, and in combination with a currently used VTE treatment. The results of this microfluidic trial will be correlated with the clinical outcome. Finally, a small group of tumours will be used to establish the optimal drug concentrations and dosing sequence for each tumour, i.e. a personalised treatment.
