2018 Fiscal Year Final Research Report

Rapid and easy detection of nephrotoxicity using human cultured cell lines; application of IL-6/IL-8 reporter gene

Research Project

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Project/Area Number	16K08548
Research Category	Grant-in-Aid for Scientific Research (C)
Allocation Type	Multi-year Fund
Section	一般
Research Field	General pharmacology
Research Institution	Tottori University
Principal Investigator	KOKURA Kenji 鳥取大学, 医学部, 助教 (30344039)
Co-Investigator(Kenkyū-buntansha)	大林徹也鳥取大学, 研究推進機構, 准教授 (80348804) 井上敏昭鳥取大学, 医学部, 准教授 (80305573)
Project Period (FY)	2016-10-21 – 2019-03-31
Keywords	腎毒性試験 / レポーター遺伝子 / 転写制御 / ヒト腎臓培養細胞株
Outline of Final Research Achievements	The development of a rapid and easy detection of nephrotoxicity using human cultured cell contributes to the reduction of the number of experimental animals used for the toxicity test and the reduction of the total cost of development of pharmaceuticals. Recently, it has been reported that in kidney-derived primary cultured cells, the expression level of IL-6 / 8 mRNA is increased in response to nephrotoxic substances. In this study, we attempted to establish a screening system using cultured human kidney cell lines into which IL-6 / IL-8 gene reporter has been introduced to detect nephrotoxicity simply and quantitatively. For this purpose, we constructed the IL-6 + IL-8 gene reporter genes, chose appropriate cultured human kidney-derived cells and confirmation of assay conditions, and examined whether the reporter gene responded to nephrotoxic substances. We finally confirmed the expected response using this reporter system with small number of test compounds.
Free Research Field	転写制御
Academic Significance and Societal Importance of the Research Achievements	腎臓は血液から老廃物を濾過後、水分や栄養分を再吸収する機能をもつため、毒性物質の暴露を受けて傷害されやすい。医薬品などの開発における腎毒性試験には、主にラットなどの実験動物が用いられてきた。しかし、実験動物で多くの化合物の腎毒性を明らかすることは費用や時間がかかるうえ、腎毒性試験を通過した医薬品候補化合物の多くが臨床試験での腎傷害により開発断念されているなどの問題がある。そのため医薬品の開発初期段階でも利用しうる、簡便な培養細胞による腎毒性試験法の併用が期待されている。本研究は、この期待に資するものであり、将来的な簡便な腎毒性試験法開発に貢献したと思われる。