14TSB_NAT A novel integrated BBB-brain model for comprehensive drug permeability and toxicity testing
Lead Research Organisation:
Queen Mary University of London
Department Name: School of Engineering & Materials Scienc
Abstract
The blood-brain barrier (BBB) is a highly organised microvasculature system composed of different cell types including brain endothelial cells, pericytes and astrocytes. Disruption of the BBB results in various brain diseases including stroke and Alzheimer's disease. Understanding of BBB integrity and maintenance is, therefore, essential to developing measures for preventing its disruption and also for effective drug delivery to the brain. Currently, most studies on BBB integrity and damage are performed on in vivo models and the current in vitro models do not reflect the cellular interactions that occur in in vivo, as different BBB cell types are cultured and studied in different types of media. The present project aims to develop a three dimensional (3D) in vitro BBB construct within the Kirkstall Quasi-Vivo culture system, which allows multiple cell types to be cultured in inter-connected chambers. Real time tracking of solutes across the BBB will be undertaken using a novel combination of laser and electrochemical biosensor based interrogation. Once developed, this in vitro model will be more accessible than an in vivo animal model for investigation of drug transport across the BBB
Technical Summary
The blood-brain barrier (BBB) is a diffusion resistant barrier with complex permeability properties mediated by both the cellular and associated extracellular biopolymer matrix. The former comprises an integrated complex of endothelial cells, pericytes and astrocytes. Baseline barrier properties and their changes in disease states serves as both a criterion of what is 'normal' and as a basis for quantitaive understanding of structure/function relationships. The Kirkstall Quasi-Vivo culture system secures a two compartment model for culture of BBB cell types and, specifically, allows correlation of cell populations with permeability. Importantly, the project will use continuous tracking electrochemical (as well as optical) sensors for monitoring selected organics/metabolites and will give quantitative diffusion coefficient data in Fickian terms rather than as a semi-quantitative index. The eventual integration of sensors and cultured cells will serve as a platform for future bioreactors for endothelial and other barrier tissue constructs. By selection of diffusants with different molecular properties (charge, polarity, size), it will be possible to establish different types of barrier function, and in addition its relation to structural changes in the BBB (ageing/disruption), including the relative balance of transcellular and paracellular transport. The 3D construct provided by Leeds University and the controlled flow system of Kirkstall will be key to meaningful data generation, especially since flow control will enable an assessment shear force effects at the endothelial surface, and thus its direct impact on cell properties as well the indirect effect on solute transport from the bulk solution. The integrated system will provide a basis for future analysis of transport asymmetry, hysterisis, saturation effects and competitive transport. A further aspect of the work will be the design of protein support layers as a basement membrane equivalent.
Planned Impact
This project supports the government agenda of the 3Rs for animal welfare and will offer both DEFRA and MHRA reliable tests to implement their animal free policies. We will inform these policy developers of options for in vitro tests to lead the requirements of new medical technologies and products at international level.
The overall society will benefit from the economic gains, as well as from the medical advances to treat neurobiological diseases, including new drugs developed using animal free platforms. Society will also gain from workforce capacity building for the training of highly qualified interdisciplinary specialised professionals.
Public perception of science and technology will also be enhanced by tangible medical benefits to society, and by involving public and patient groups to provide feedback.
The overall society will benefit from the economic gains, as well as from the medical advances to treat neurobiological diseases, including new drugs developed using animal free platforms. Society will also gain from workforce capacity building for the training of highly qualified interdisciplinary specialised professionals.
Public perception of science and technology will also be enhanced by tangible medical benefits to society, and by involving public and patient groups to provide feedback.
People |
ORCID iD |
Pankaj Vadgama (Principal Investigator) |
Publications
Adatia K
(2017)
An electrochemical study of microporous track-etched membrane permeability and the effect of surface protein layers.
in Colloids and surfaces. B, Biointerfaces
Description | Diffusion through a protein barrier is retarded and the pore area of membranes does not correlate simply or directly with their diffusive permeability. So for cell culture such membranes should be assessed for their barrier properties and reliance on structure is not predictive of performance on diffusive resistance. Work by a medical student linked to the project led to the finding of highly reduced diffusion in micropores - this breaks with assumptions and indicates a change in water structure. The work will be the basis of a grant application to EPSRC. |
Exploitation Route | The use of sensors for bioreactor monitoring generally and the use of software that simplifies the assessment of biopolymer and packaging material permeability to solutes and gases. |
Sectors | Agriculture Food and Drink Healthcare Manufacturing including Industrial Biotechology Pharmaceuticals and Medical Biotechnology |
Description | Queen Mary, University of London PhD Studentship |
Amount | £56,000 (GBP) |
Organisation | Queen Mary University of London |
Sector | Academic/University |
Country | United Kingdom |
Start | 08/2016 |
End | 08/2019 |
Title | Miniature biosensors |
Description | Electrochemical miniature sensors devised for metabolite tracking across a cell/epithelial barrier layer. Simple measurement of mass transport through biopolymer and other barriers using electrochemical sensors. |
Type Of Material | Technology assay or reagent |
Provided To Others? | No |
Impact | Setting up of an SME for possible development;the SME is Camstech Ltd and has funding from CERN for innovation development of fsensors. |
Description | Membrane materials for implantable biofuel cells |
Organisation | University of Grenoble |
Country | France |
Sector | Academic/University |
PI Contribution | Design of experiments for biofuel cells. External supervisuon of PhD student and PDRA |
Collaborator Contribution | Provision of laboratory resources and facilities to test out ideas and joint supervision of junior researchers. Also, funding for visiting Professorship for 4 months |
Impact | One paper submitted to a journal. One paper in preparation. Patent under consideration. One review at revision stage. One review in preparation. |
Start Year | 2017 |
Description | Advances in Cell and Tissue Culture Pisa |
Form Of Engagement Activity | A talk or presentation |
Part Of Official Scheme? | No |
Geographic Reach | International |
Primary Audience | Professional Practitioners |
Results and Impact | Presentation of work as poster showing slowed drug transport through a protein layer. Title: A modified polymer membrane support for an in vitro blood brain barrier system. Showed a cell biology audience the way electrochemistry might be used. |
Year(s) Of Engagement Activity | 2015 |
Description | Intercalated BSc Symposium at Queen Mary University of London |
Form Of Engagement Activity | A talk or presentation |
Part Of Official Scheme? | No |
Geographic Reach | Local |
Primary Audience | Undergraduate students |
Results and Impact | 55 students + staff attended a series of intercalated BSc students presentations. The work was presented as "Anomalous, Low Diffusion Through Micropores: Implications for Mass Transport Control" by a student working within the project. Led to questions on the experimental protocol used. |
Year(s) Of Engagement Activity | 2015 |