日本質量分析学会 第66回質量分析総合討論会

Program

Oral Sessions

Day 4, May 18(Fri.) 10:00-10:15 Room B (Seiun 1)

Identification of internalized plasma membrane proteins in human brain capillary endothelial cells.

(Kumamoto Univ.)
oMariko Oishi

The blood-brain barrier (BBB) mainly consists of brain capillary endothelial cells. The BBB restricts the transport of therapeutic drugs, so efficient receptor-mediated transcytosis (RMT) across the BBB is an important strategy for drug delivery to the central nervous system. Thus, the purpose of the present study is to identify novel cell surface proteins selectively expressed and internalized into human brain capillary endothelial cells in vitro for identification of novel RTM molecules at the BBB. To find BBB selective RMT molecules, we performed a cell surface biotinylation-based internalization method on BBB model cells and other peripheral blood vessel model cell. Silver staining showed that biotinylated proteins were detected in the cell surface and internalized fraction. Using SWATH-MS analysis, we identified proteins in the cell surface fractions. Among these, we estimated the biotinylated proteins after excluding the cytosolic proteins annotated via Gene Ontology analysis. Furthermore, BBB-selective internalized proteins were extracted using our original criteria. Considering the expression levels of these proteins in the isolated hCMEC/D3 plasma membrane fraction using SWATH-MS, 20 proteins were speculated to be candidate molecules for RMT at the BBB. Our present study provides novel information regarding the development of a BBB permeable, brain-targeted drug delivery system.