Development and assessment of new risk assessment methods

Title of Research:
17_S05-01

Development of Novel Analytical System of Biological Responses toChemicals by Deep Phenotyping Method
Principal Investigator:
Hiroyuki Kusuhara (Professor, University of Tokyo, Graduate School of Pharmaceutical Sciences,
Department of Molecular Pharmacokinetics)
7-3-1 Hongo, Bunkyo, Tokyo, 113-0033, Japan(Tel) +81-3-5841-4770, (E-mail) kusuhara@mol.f.u-tokyo.ac.jp
Collaborators:
Tadahaya Mizuno (Assistant Professor, University of Tokyo, Graduate School of PharmaceuticalSciences, Department of Molecular Pharmacokinetics)7-3-1 Hongo, Bunkyo, Tokyo, 113-0033, Japan(Tel) +81-3-5841-4771, (E-mail) tadahaya@mol.f.u-tokyo.ac.jp
Setsuo Kinoshita (Contract Research Student, University of Tokyo, Graduate School of
Pharmaceutical Sciences, Department of Molecular Pharmacokinetics)
7-3-1 Hongo, Bunkyo, Tokyo, 113-0033, Japan
Summary of Research:
Today, organisms are always exposed to chemicals in every situation to some extent. Wide varietyof chemicals exist around us and the chemicals affect organisms not one by one but
simultaneously like a complex. The fact prompts us to care “multiple exposure”. To achieve theappropriate and correct prediction of multiple exposure of chemicals, it is necessary tocomprehend the effects of chemicals even if those are hidden and difficult to be detected. In thisstudy, we tackle with the theme by omic approaches with innovative techniques in both dataacquisition and data analysis method.
This study consists of two pillars: (1) a novel acquisition method of proteome profile data by
2-dimensional electrophoresis (2DE) and (2) a new analysis method of profile data. In the 5th term,
we carried out fine-tuning of these two.
As for 2DE, we have investigated intra-day deviation of profile data because it is indispensablefor the construction of database. HeLa cells treated with or without bortezomib were lysed and thespecimens were subjected to 2DE. The difference between days were smaller than that between
treatments (bortezomib and DMSO as a control). We have written the detailed protocol of our 2DE
method to minimize the dependency of the data on investigators.
As for data analysis, we have summarized the results so far and it was accepted by ScientificReports. By this method, it is expected that profile data of samples subjected to multiple exposures
can be separated in unsupervised manner, and it can be estimated what kind of action iscombined. In this term, we searched for data subjected to combined exposure from Gene
expression omnibus (GEO; https://www.ncbi.nlm.nih.gov/gds), and analyzed the data analysis with
OLSA. Six combined exposure data were found (ID: GSE10466, GSE16816, GSE17589,GSE24065, GSE35230, GSE59882). Of these, combined exposure data of GSE16816,GSE24065, and GSE59882 were expressed in a linear combination of vectors obtained by OLSA.
Timeline:
March 1, 2018 – February 28, 2019
Topics:2018 LRI annual meeting, “Development of Novel Analytical System of Biological Responses toChemicals by Deep Phenotyping Method”
Publications:Akademi) 第 ２ 回 日 韓 若 手 薬 剤 学 研 究 者 ワ ー ク シ ョ ッ プ , Seul, 2018/7/9-11; GPEN2018,

Development and assessment of new risk assessment methods
Singapore, 2018/9/25-29; 33rd JSSX, Ishikawa, 2018/10/1-5; CBI 2018, Tokyo, 2018/10/9-10/11;
第 40 回生体膜と薬物の相互作用シンポジウム, 2018/10/18-19, Miyagi; THE 13th SUGIYAMALABORATORY (RIKEN) OPEN SYMPOSIUM, 2019/2/22, YokohamaJournal) Mizuno T, Sci Rep, 2019