日本質量分析学会 第66回質量分析総合討論会

Program

Poster Presentations

Day 4, May 18(Fri.)  Poster

Protein Biomarker Discovery in Extracellular Vesicles isolated from Plasma for Colorectal Cancer

(1NIBIOHN, 2Kyoto Univ., 3QBIC, 4Jpn. Fdn. Cancer Res.)
oKeiko Kasahara1,2, Ryohei Narumi1, Yoshihiro Shimizu3, Keiko Masuda3, Yuichi Abe1, Jun Adachi1, Satoshi Nagayama4, Yoshiharu Sakai2, Takeshi Tomonaga1

The recent development of mass-spectrometry-based technologies has been providing unprecedented insights in biomarker discovery. However, it is still regarded as extremely challenging to provide high-reproducibility in plasma proteome analysis for a small amount of protein. Isolating extracellular vesicles (EVs) from plasma is one of ideal methods to overcome this problem in the point of reducing abundant proteins. We attempted to gradually narrow down biomarker candidates (BCs). We isolated EVs from plasma in colorectal cancer (CRC) patients (n=59) and healthy controls (n=59) by using ultracentrifugation. LC-MS/MS was used to provide an overview of plasma EVs protein profile (Dataset 1). Primary BCs were selected by three datasets: PubMed literature search for proteins that had been reported to be functionally correlated to the pathogenesis of CRC (Dataset 2); ExoCarta database research for proteins that had been reported their expression in EVs (Dataset 3). LC-SRM/MS was used to obtain absolute quantification of primary BCs for selection of secondary BCs. MS-based Quantification By isotope-labeled Cell-free products (MS-QBiC) 1) provided high-throughput preparation of internal standards. As validation phase, we used isolated EVs from plasma in CRC patients (n=50) and healthy controls (n=50) to evaluate the performance of secondary BCs. Eventually, novel biomarker were identified.