A microfluidic platform for modeling drug transport and cell trafficking across the blood-brain barrier

DESCRIPTION provided by applicant The blood brain barrier BBB is a tight barrier formed by microvessels and capillaries that control the passage of nutrients fluids metabolic products and drugs between the blood and the brain Impaired function of the BBB is involved in a number of major pathologies afflicting the brain such as Alzheimerandapos s disease multiple sclerosis Parkinsonandapos s disease brain manifestations of AIDS stroke and cancer Although the neurotherapeutics sector is among the largest and fastest growing markets in the pharmaceutical industry progress is currently impaired by the lack of in vitro assays that reliabl predict in vivo BBB permeability None of the existing models adequately replicates the organotypic microenvironment of the BBB in which brain endothelial cells ECs pericytes PCs and astrocytes ACs are arranged in a characteristic architecture The proposed work utilizes organ on chip technology recently developed by Nortis Inc for creating D tissue microenvironments in disposable microfluidic chips The chip design enables the integration of living lumenally perfused microvasculature making it suitable for studying barrier function Strikingly extensive preliminary data indicate that human brain ECs PCs and ACs have the capacity to self assemble into a BBB like architecture within the Nortis chip This data will be leveraged to further develop and eventually commercialize BBB models of mouse and human The objective of Phase I is to achieve a model that replicates critical BBB functions of the mouse brain The mouse model will be developed and optimized for viability structure and function Expression of tight junction TJ proteins and the transporter P glycoprotein an important functional characteristic of the BBB will be measured Microvessel permeability will be assessed by perfusion with fluorescently labelled molecules Aim The model will then be challenged with the barrier modulating compound lipopolysaccharide LPS and evaluated for associated changes in TJ protein expression molecule permeability and leukocyte transendothelial migration Aim During Phase II the mouse BBB chip will be used to develop and qualify specific BBB assays such as transferrin receptor transporter activity BBB permeability challenge with LPS and stimuli induced leukocyte transmigration Aim Success criteria is an assay robustness of Zandapos Aim of Phase II is to develop a human BBB model The human model will be optimized to recapitulate key structural and functional features of the BBB including TJ formation permeability and transporter activity To demonstrate utility the model will be treated with LPS mannitol and angiotensin II and evaluated for associated changes in BBB structure and function Each of these compounds has clinical relevance but acts by a different mechanism Aim is to qualify specific human BBB assays and establish relevance to clinical data The products developed with support from this grant will significantly enhance progress in basic translational and clinical neuroscience research and will significantly advance therapy for numerous devastating diseases PUBLIC HEALTH RELEVANCE Impaired function of the blood brain barrier contributes to a number of diseases including Alzheimerandapos s disease multiple sclerosis Parkinsonandapos s disease malignancies of the brain and stroke The high failure rate of drugs targeting these disorders highlights the critical need for blood brain barrier models that better predict clinical outcomes Here microfluidic technology is utilized to develop two new in vitro models that replicate a number of key in vivo barrier functions providing an important alternative to current in vitro models and animal testing