2020 Fiscal Year Final Research Report

Development and application of disease-specific protein targeting system

Research Project

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Project/Area Number	19K22564
Research Category	Grant-in-Aid for Challenging Research (Exploratory)
Allocation Type	Multi-year Fund
Review Section	Medium-sized Section 50:Oncology and related fields
Research Institution	Ehime University
Principal Investigator	Higashiyama Shigeki 愛媛大学, プロテオサイエンスセンター, 教授 (60202272)
Co-Investigator(Kenkyū-buntansha)	小川敦司愛媛大学, プロテオサイエンスセンター, 准教授 (30442940) 福田信治愛媛大学, プロテオサイエンスセンター, 講師 (70398238)
Project Period (FY)	2019-06-28 – 2021-03-31
Keywords	DNA aptamer / protein targeting / SPOP / CUL3 / CBF1
Outline of Final Research Achievements	Mutated proteins are promising therapeutic targets in various diseases including cancers. Bi-functional protein-targeting chimeric molecules PROTACs which cross-link ubiquitin (Ub) E3 ligases and non-substrate proteins have been developed. However, successfully developed PROTACs are quite limited, because they consist of chemical compounds. On the other hand, it has been shown that single strand DNA aptamers are able to recognize specifically even single amino-acid mutated proteins. In this study, we tried to develop disease specific protein targeting modulators based on CUL3-SPOP and DNA aptamers. We successfully identified a SPOP-binding chemical compound #14607 and isolated 15 kinds of DNA aptamers for CBF1 (a target protein) with high affinity (Kd: 10-300 nM), and developed CBF1 targeting modulators, #14607-CBF1-DNA aptamers. We tested them in an in vitro Ub-lation assay system. Unfortunately, we have not detected CBF1 Ub-lation in this system yet.
Free Research Field	生化学、細胞生物学
Academic Significance and Societal Importance of the Research Achievements	遺伝子解析技術の進歩は、様々な疾患特異的遺伝子変異を明らかにしてきているが、治療目的の個体内で遺伝子修復は極めて遠い道のりである。では、「遺伝子変異によって産生される変異タンパク質を標的とする治療薬開発は可能か?」というと、個々の変異タンパク質に対して個別に創薬を進める必要があることから、極めて困難な道筋である。しかし、個々の変異タンパク質に対する創薬に、一つの共通技術プラットホームができれば、この戦略は飛躍的に前進する。本研究の成果は、その共通技術プラットホームの創出を可能にするものであり、従来法より作成時間と労力を大幅に短縮でき、その社会的意義は極めて大きい。