In vitro modeling of blood brain barrier dysfunction on a chip to elucidate the pathogenesis of cerebral malaria

Brain microvascular endothelial cells (BMECs) which line the vascular network of the central nervous system (CNS) in conjunction with perivascular cells form a specialized barrier termed the blood-brain barrier (BBB) that regulates the dynamic traffic of select molecules into and out of the CNS. Studies of BBB dysfunction have been hampered by an inability to perform direct testing in patients and a lack of in vitro models. Analysis of available putative BMECs from pluripotent stem cell sources reveals that they do not harbor bona fide brain EC molecular signatures. Using a transcription factor-based reprogramming strategy, it was demonstrated that durable and functional true endothelial cells with BBB traits may be derived. These verified BMECs will be used in the development of isogenic BBB/on-a-chip devices. Cortical brain organoids derived from patient specific induced pluripotent cells (IPSC) will be vascularized by matched IPSC derived bona fide BMECs in a microfluidic device.

Red blood cells infected with the Plasmodium falciparum parasite adhere to the brain microvasculature causing dysfunction. It is hypothesized that the parasite primes the brain endothelial cells for adherence through their “education” by small extracellular vesicles termed exosomes. This is an established paradigm in cancer biology in which malignant cells educate metastatic “microniches” to prime for and enhance tumor survival. The exosomes mediate local and systemic intercellular communication through the horizontal transfer of information in their proteomes. Recently a critical prognosis biomarker for brain metastasis in breast cancer through studies of exosomal cellular uptake and proteomic analysis was identified. The proteomes of exosomes from children with cerebral malaria will be analyzed and infuse them into our innovative microfluidic-based BBB model. How exosomes trigger BBB dysfunction and subsequently affect IPS-derived brain organoids will be monitored in real time from patients affected by CM. These studies will provide new insights into disease pathogenesis and potentially identify new prognosis biomarkers.

Project details

Basic Science
Product Development

Sep 2021 — Aug 2023

$1.68M

United States