Method establishment of next generation clinical diagnosis based on highly precise pathological specimen proteomics

2020 Fiscal Year Final Research Report

• Project/Area Number: 19K22590
• Research Category: Grant-in-Aid for Challenging Research (Exploratory)
• Allocation Type: Multi-year Fund
• Review Section: Medium-sized Section 52:General internal medicine and related fields
• Research Institution: Tohoku University
• Principal Investigator: Uchida Yasuo 東北大学, 薬学研究科, 講師 (70583590)
• Project Period (FY): 2019-06-28 – 2021-03-31
• Keywords: 臨床診断 / 病理学 / ホルマリン固定パラフィン包埋（FFPE） / Advanced SWATH法 / プロテオミクス / PCT / 熱 / PTS

Outline of Final Research Achievements

The purpose of this study was to establish quantitative proteomic method able to accurately quantify pathological changes in the protein expression levels using FFPE samples. After Pressure Cycling Technology (PCT)-assisted processing of FFPE liver samples followed by SWATH-MS-based comprehensive quantification, the peak areas of 88.4% of peptides agreed with those from matched fresh samples within a 1.5-fold range. The present method using phase-transfer surfactant (PTS) buffer at 95C showed better agreement of peptide peak areas between FFPE and fresh samples. When our method using PCT and PTS buffer at 95C was applied to a bile duct ligation (BDL) disease model, the BDL/control expression ratios for 80.0% of peptides agreed within 1.2-fold range between FFPE and fresh samples. This heat-compatible FFPE-PCT-SWATH proteomics technology using PTS is suitable for quantitative studies of pathological molecular mechanisms and biomarker discovery utilizing widely available FFPE samples.

Free Research Field

臨床診断学

Academic Significance and Societal Importance of the Research Achievements

臨床診断において、病変組織におけるタンパク質の“定量”は欠かせない技術である。しかし、日常的な病理診断に利用されているFFPE組織に対して有用な定量法は確立されておらず、極めて挑戦的な課題とされてきた。本研究では、FFPE検体で利用可能な、高精度かつ網羅的なタンパク質定量技術を開発し、世界で初めてFFPE組織を使って病態分子機構を定量的に解析できることを実証した。従って、本研究は、FFPE組織の利用価値を飛躍的に高めた点で、学術面だけでなく、臨床的・社会的に重要な意義をもつものと考えらえる。