Executive Summary : | This project aims to design effective peptide molecules for drug delivery into the brain, as drug delivery is challenging due to the blood-brain barrier (BBB). In the case of meningitis, Escherichia coli K1 (E. coli K1) can cross the BBB and enter the brain compartment without affecting the integrity of the BBB. The project proposes a different approach to solve the brain drug delivery problem by designing peptide molecules from OmpA that activate protein kinase C (PKC)-α and rearrange the actin cytoskeleton framework of BMEC, increasing the permeability of the BBB. The project consists of four parts: designing and synthesizing OmpA-derived peptide molecules, investigating the peptide-Ecgp96 receptor interaction and uptake of peptide molecules into BMEC, testing the effect of these peptide molecules on the actin cytoskeleton of BMEC and activation of PKC-α, and checking the permeability enhancement due to peptide treatment across a model BBB made with a BMEC monolayer. OmpA binds with the Ecgp96 receptor on the BMEC's surface through its amino acids of four extracellular loops. The peptide molecules will be designed based on the involvement of amino acids of OmpA with Ecgp96. OmpA-derived peptide molecules will be synthesized and labeled with 5(6) carboxyfluorescein (FAM) molecules to study the cellular uptake of peptides in BMEC. A cellular uptake experiment will identify the best peptide sequence targeting the Ecgp96 receptor and entering the BMEC. Finally, the permeability of the peptide treated and control BMEC monolayer will be checked using horseradish peroxidase (HRP) transport across the confluent monolayer of BMEC in a transwell system. |