14TSB_NAT Neurotox: A novel integrated BBB-brain model for comprehensive drug permeability and toxicity testing

Lead Research Organisation: University of Leeds
Department Name: School of Medicine

Abstract

The blood-brain barrier (BBB) is composed of different specialised cell types and regulates exchange of substances between the blood and the brain. There are a large number of diseases including stroke, brain trauma and tumours in which the permeability of the BBB is increased. Conversely, many drugs are unable to cross the BBB to reach the brain making the BBB one of the major obstacles in the treatment of the brain diseases. Proper regulation and maintenance of BBB is, therefore, essential for effective drug delivery to the brain to cure brain diseases and preventing its further damage. In this project we aim to develop a three dimensional in vitro BBB model using the Kirkstall Quasi-Vivo system which allows multiple cell types to be cultured in inter-connected culture chambers and a method of assaying the passage of of potential drugs across the BBB by electrochemical and laser bio-sensors. This in vitro model will be an important tool for investigation of different aspects of BBB function and testing potential drugs for their permeability and toxicity and reducing animal usage in this area. Human cell based models should be more accurate and predictive than animals.

Technical Summary

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 bio-sensor 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.

Planned Impact

Please see the application submitted by the TSB partner
 
Description We have developed an in vitro multi-cellular model of by culturing human primary cell types forming the blood brain barrier (BBB) namely, brain primary endothelial cells, pericytes and astrocytes within the Kirkstall Quasi Vivo® 500 system. This system allows multiple cell types to be cultured in inter-connected chambers sharing the same medium by all cells under flow. Optimal medium and flow rates for growing all three cell types were established to define the best conditions for developing an in vitro dynamic BBB model in the future. As a proof of concept the effects of amyloid-beta 25-35 peptide (Aß25-35), a hallmark of Alzheimer's Disease, were explored on BBB individual cell types grown together under the same conditions. We also co-cultured the three human primary main cell types involved in BBB formation to create a human static BBB system.Tightness of thec barrier and permeability are studied by measuring cellular resistance and passage of molecules of different sizes and properties.
Exploitation Route This multi-cellular model will be suitable for studying the role of each BBB cell type separately in CNS disorders as well as new targets for these diseases. The finding can be used by academics for understanding transport mechanisms through the blood brain barrier in health and diseases and also by the pharmaceutical companies for drug delivery and toxicity testing. WE have published the work in Scientific report. .
New sensor development and use of Nano particles for bio-imaging have been facilitated.
Sectors Chemicals,Construction,Digital/Communication/Information Technologies (including Software),Education,Healthcare,Pharmaceuticals and Medical Biotechnology

URL https://www.nature.com/articles/s41598-018-26480-8
 
Description The findings have been used for further grant funding and for publications. The organ flow system model developed in this project has been used in BBSRC-TDRF grant led by Professor Paul Millner in the Faculty of Biological Sciences and an EU Marie Curie fellowship for Dr Padmaje Vasudev. I'm acting as co-I in both these proposals. The findings were also used for conference presentations e.g., Society for Neuroscience, USA, British Society for Cardiovascular Research . The Kirkstall Ltd have used the findings for commercialisation, business plan and setting up meetings with pharmaceutical companies. The findings are being used for further grant funding including EU funding and for industrial partnership. The findings were presented as part of my invited lecture at the National Brain Centre, Maneswar, Hariyana, India and the UK-China Neuroprotection Research Platform Conference" in Bradford, UK. I organised a symposium on Blood brain barrier at the 23rd Biennial Meeting of the European Society NeuroScience, September 1-4, 2019, Milan, Italy Guest Editor, special edition on Cerebrovascular Diseases, Frontiers in Cellular Neuroscience 2018 ( doi: 10.3389/978-2-88963-442-2) New publications on blood brain barrier system and bioimaging have been made,1) https://www.nature.com/articles/s41598-018-26480-8 (DOI:10.1038/s41598-018-26480); 2) https://onlinelibrary.wiley.com/doi/full/10.1002/jbio.201700199 (doi: 10.1002/jbio.201700199); 3) https://www.ncbi.nlm.nih.gov/pubmed/30350344 (DOI: 10.1002/jbio.201800256) The data has also been used in our recently awarded EPSRC grant (starting in 1st April 2020) in which I'm a Co-I.
First Year Of Impact 2017
Sector Chemicals,Construction,Education,Healthcare
Impact Types Societal,Economic

 
Description BBSRC project grant
Amount £500,000 (GBP)
Organisation Biotechnology and Biological Sciences Research Council (BBSRC) 
Sector Public
Country United Kingdom
Start 04/2016 
End 03/2019
 
Title BBB model 
Description Developed a blood brain barrier model under flow 
Type Of Material Model of mechanisms or symptoms - in vitro 
Year Produced 2015 
Provided To Others? Yes  
Impact Developed new grant applications 
 
Description BLDE University, India. 
Form Of Engagement Activity A formal working group, expert panel or dialogue
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact Delivered a talk to 150 undergraduate students
Delivered a seminar to approx. 200 academics/ professionals
Year(s) Of Engagement Activity 2016
 
Description Plenary lecture 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact About 200 delegated attended the conference, people from defense institute in India also were in the audience. Discussion were sparked afterwards on neuro-degenerative diseases
Year(s) Of Engagement Activity 2017