In Vitro Organ Imaging Device (IV-OID) with integrated Biosensing and Real-Time Imaging Capability: Proof-of-Principle using a Human Placental Model
Lead Research Organisation:
University of Liverpool
Department Name: Clinical Infection, Microbiol & Immun
Abstract
Bringing a new drug or a vaccine to the market is a complex task for both researchers and drug manufacturers. Recent studies reported that the costs of developing a drug had nearly doubled every decade between 1950 and 2010. Today's drug or vaccine candidates are more likely to fail in clinical trials compared to 40 years ago. Different reasons have been put forth explaining the high failure rate, one of them being that preclinical models cannot fully recapitulate the human physiology. Hence, the need for more complex and predictive in vitro models which can better mimic the human response to drug or vaccine is essential.
Here, we propose to develop a novel technological platform i..e, an In Vitro Organ Imaging Device (IV-OID) based on the use of hollow fibre technology and imaging-compatible materials. The platform aims to address the following objectives:
(a) Creation of a human tissue surrogate platform that researchers can use e.g., to test drugs, in lieu of animals or humans.
(c) Engineering of an organ-like model systems as a means of providing insight into the mechanisms underlying infectious diseases.
(b) High-resolution visualisation of how pathogens behave and lead to diseases using state-of-the art microscopes.
Our technological platform will be validated through the set up of a human placental model and the evaluation of its barrier functions in relation to microbial pathogens such as Toxoplasma Gondii. If successful, the IV-OID will be made available to the wider academic and industrial communities through our industrial project partner, Flocel Inc.
Here, we propose to develop a novel technological platform i..e, an In Vitro Organ Imaging Device (IV-OID) based on the use of hollow fibre technology and imaging-compatible materials. The platform aims to address the following objectives:
(a) Creation of a human tissue surrogate platform that researchers can use e.g., to test drugs, in lieu of animals or humans.
(c) Engineering of an organ-like model systems as a means of providing insight into the mechanisms underlying infectious diseases.
(b) High-resolution visualisation of how pathogens behave and lead to diseases using state-of-the art microscopes.
Our technological platform will be validated through the set up of a human placental model and the evaluation of its barrier functions in relation to microbial pathogens such as Toxoplasma Gondii. If successful, the IV-OID will be made available to the wider academic and industrial communities through our industrial project partner, Flocel Inc.
Technical Summary
The most widely used in vitro models for human tissue-blood barriers are based on static setups such as the Transwell system, which poorly mimic physiological conditions, hence have limited translational significance. While efforts have been made towards the development of more predictive models, including that of a human trophoblast organoid, key limitations resides in (1) their inability to recreate the dynamic flow exchange existing at the placental barrier, or (2) their inability to replicate the mature developmental structure of the placenta, and, (3) the complexity of setting up the models in a research lab.
Owing to the ethical issues associated with the study of human pregnancy and the caveat inherent to current experimental models, the understanding of the mechanisms governing placenta-blood barrier functions remains limited. The human placenta is an intricate organ made up of cellular and vascular networks that act in concert to promote fetal growth and viability. Trophoblasts represent the most outer layer of the placenta whereby chemical and gas feto-maternal exchanges are regulated. Here we propose to develop an unprecedented trophoblast-based microfluidic dynamic model, that mimics the feto-maternal interface. We will build on the use of hollow fibre technology to develop an artificial capillary system that will provide a three-dimensional, quasi-physiological environment where (fetal) endothelial cells will be luminally exposed to physiological levels of flow and co-cultured with abluminal trophoblasts to form a functional placental barrier. Our platform will be unique in that it will also offer real-time multi-photon imaging capability.
Our easy-to-set up 3D-dynamic flow model may be used in a variety of research applications encompassing elucidation of vertical disease transmission mechanisms e.g., TORCH pathogens, maternal antibody placental transfer, pharmacodynamics of antibiotics and drugs.
Owing to the ethical issues associated with the study of human pregnancy and the caveat inherent to current experimental models, the understanding of the mechanisms governing placenta-blood barrier functions remains limited. The human placenta is an intricate organ made up of cellular and vascular networks that act in concert to promote fetal growth and viability. Trophoblasts represent the most outer layer of the placenta whereby chemical and gas feto-maternal exchanges are regulated. Here we propose to develop an unprecedented trophoblast-based microfluidic dynamic model, that mimics the feto-maternal interface. We will build on the use of hollow fibre technology to develop an artificial capillary system that will provide a three-dimensional, quasi-physiological environment where (fetal) endothelial cells will be luminally exposed to physiological levels of flow and co-cultured with abluminal trophoblasts to form a functional placental barrier. Our platform will be unique in that it will also offer real-time multi-photon imaging capability.
Our easy-to-set up 3D-dynamic flow model may be used in a variety of research applications encompassing elucidation of vertical disease transmission mechanisms e.g., TORCH pathogens, maternal antibody placental transfer, pharmacodynamics of antibiotics and drugs.
Planned Impact
Staff
Staff working on the research project will develop skills that will be transferrable to other sectors of the employment market including communication and interpersonal skills, organization, record keeping and statistical analysis skills. The proposal will involve a period of training within the premises of our industrial partner Flocel Inc. The post-doctoral scientist employed on the project will have the opportunity to travel there, an invaluable opportunity to develop new skills and enhance future career prospects. The research staff would also develop laboratory skills in a number of key areas as the proposal is largely multi-disciplinary i.e., fluidics, fluorescence imaging and infectious disease.
Academics
This proposal will develop new and improved collaborations both within the UK (UoLiverpool with Liverpool Womens hospital and UoManchester) and abroad (Liverpool with As, Norway, Portugal and USA). These collaborations will be across multidisciplinary research fields and will develop knowledge and skill sets for all those involved.
Within the University of Liverpool, establishing a link between microbiologists at the Institute of Infection and Global Health and clinicians at the Liverpool Women's Hospital will broaden knowledge for all parties involved. The novel combination of human in vitro cell culture, clinical studies and in vivo infection models will open up collaboration opportunities in the UK and abroad and data generated from the proposal will be used to secure further funding for the University through research grant applications.
The primary focus of the project is imaging, microbiology and immunology, and so academics in these areas will be the primary beneficiaries. Although the proposal will use Toxoplasma gondii a model pathogen, the approaches used and the interactions uncovered are likely to be of direct relevance to those working on other TORCH pathogens e.g., cytomegalovirus.
Clinicians, Healthcare Professionals and Industry
Data from this project will be of direct relevance to clinical researchers working on the preventive control of congenital disease. In the longer term, based on the findings of our project, academic researchers, industry and clinicians will benefit from adopting a novel in vitro tool towards more translational research.
The Public
Within the duration of the grant we would benefit the public through promoting wider appreciation of the use of 3D in vitro model platforms. This would be achieved through publication of research findings in open access journals and with accompanying press releases in local and national press. Furthermore, talks, seminars and activity days organised at schools and in the community (libraries, museums etc.) would be arranged through the outreach organising committee within the Institute of Infection and Global Health. The proposal focuses on the creation of a model that will allow significant versatility in its applications e.g., effect of drug administration during pregnancy. These applications are likely to be of particular interest to the public and so will be the focus of much of our outreach and media work.
3Rs guiding principles.
Animal experimentations are still heavily required to investigate the causes of diseases, predict the activity and toxicity of drugs on human organs, or develop new vaccination approaches. While animal testing raises important ethical concerns, it is largely recognised today that it is in the best interest of everyone to replace and reduce animal testing as much as possible. 3D cell culture platforms such as ours promise to be a feasible alternative. Our model device has the potential to be streamlined in the drug or vaccine discovery pipeline hence accelerate drug discovery while reducing and replacing animal testing.
Further details of the above and other impact activities can be found in the attached 'Pathways to Impact' file.
Staff working on the research project will develop skills that will be transferrable to other sectors of the employment market including communication and interpersonal skills, organization, record keeping and statistical analysis skills. The proposal will involve a period of training within the premises of our industrial partner Flocel Inc. The post-doctoral scientist employed on the project will have the opportunity to travel there, an invaluable opportunity to develop new skills and enhance future career prospects. The research staff would also develop laboratory skills in a number of key areas as the proposal is largely multi-disciplinary i.e., fluidics, fluorescence imaging and infectious disease.
Academics
This proposal will develop new and improved collaborations both within the UK (UoLiverpool with Liverpool Womens hospital and UoManchester) and abroad (Liverpool with As, Norway, Portugal and USA). These collaborations will be across multidisciplinary research fields and will develop knowledge and skill sets for all those involved.
Within the University of Liverpool, establishing a link between microbiologists at the Institute of Infection and Global Health and clinicians at the Liverpool Women's Hospital will broaden knowledge for all parties involved. The novel combination of human in vitro cell culture, clinical studies and in vivo infection models will open up collaboration opportunities in the UK and abroad and data generated from the proposal will be used to secure further funding for the University through research grant applications.
The primary focus of the project is imaging, microbiology and immunology, and so academics in these areas will be the primary beneficiaries. Although the proposal will use Toxoplasma gondii a model pathogen, the approaches used and the interactions uncovered are likely to be of direct relevance to those working on other TORCH pathogens e.g., cytomegalovirus.
Clinicians, Healthcare Professionals and Industry
Data from this project will be of direct relevance to clinical researchers working on the preventive control of congenital disease. In the longer term, based on the findings of our project, academic researchers, industry and clinicians will benefit from adopting a novel in vitro tool towards more translational research.
The Public
Within the duration of the grant we would benefit the public through promoting wider appreciation of the use of 3D in vitro model platforms. This would be achieved through publication of research findings in open access journals and with accompanying press releases in local and national press. Furthermore, talks, seminars and activity days organised at schools and in the community (libraries, museums etc.) would be arranged through the outreach organising committee within the Institute of Infection and Global Health. The proposal focuses on the creation of a model that will allow significant versatility in its applications e.g., effect of drug administration during pregnancy. These applications are likely to be of particular interest to the public and so will be the focus of much of our outreach and media work.
3Rs guiding principles.
Animal experimentations are still heavily required to investigate the causes of diseases, predict the activity and toxicity of drugs on human organs, or develop new vaccination approaches. While animal testing raises important ethical concerns, it is largely recognised today that it is in the best interest of everyone to replace and reduce animal testing as much as possible. 3D cell culture platforms such as ours promise to be a feasible alternative. Our model device has the potential to be streamlined in the drug or vaccine discovery pipeline hence accelerate drug discovery while reducing and replacing animal testing.
Further details of the above and other impact activities can be found in the attached 'Pathways to Impact' file.
