Development of single-cell proteomics system for accelerating drug discovery in sickle cell anemia

2018 Fiscal Year Final Research Report

• Project/Area Number: 17K15042
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: Genome biology
• Research Institution: Kumamoto University
• Principal Investigator: Masuda Takeshi 熊本大学, 大学院生命科学研究部(薬), 助教 (70383940)
• Project Period (FY): 2017-04-01 – 2019-03-31
• Keywords: 1細胞 / プロテオミクス / 油中液滴 / 高感度分析 / 質量分析

Outline of Final Research Achievements

Single cell protein analysis is a key technology for understanding of tissue and cell heterogeneity. Mass spectrometry based proteomics is possible to comprehensively identify and quantify proteins. However, it is hardly possible to perform single cell proteomics due to low sample recovery by adsorption loss during sample preparation. To reduce the contact area of a sample solution with a plastic tube, we have developed a water droplet in oil (WinO) protocol. The number of identified peptides and signal intensity in WinO were increased 1.7-fold than those in In-solution protocol. In addition, the peptide recovery was improved by optimization of protease amount, organic solvent, beads and component of surfactants in water droplet.

Free Research Field

プロテオミクス

Academic Significance and Societal Importance of the Research Achievements

同一細胞群内における分子レベルの不均一性が報告されている。細胞の個性は、幹細胞から多様な細胞種に分化を促すなど、生命活動には欠かすことができない。一方、細胞の個性は腫瘍細胞に抗がん剤耐性を付与しガンの根治を難しくするなど、人類が生きる上でネガティブな一面も持ち合わせている。タンパク質は生命活動の中心的役割を担う。1細胞プロテオミクスを実現できれば、赤芽球分化様式の解明だけでなく、腫瘍細胞集団内の抗がん剤耐性細胞やガン幹細胞など希少細胞集団に効果的な薬剤の開発、ヒト造血幹細胞の増殖法の開発など幅広い分野に利用されることが期待される。