Research on defects in ketone body metabolism: Establishment of defective cell lines and expression systems of mutant enzymes .

2018 Fiscal Year Final Research Report

• Project/Area Number: 16K09962
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Pediatrics
• Research Institution: Gifu University
• Principal Investigator: Fukao Toshiyuki 岐阜大学, 大学院医学系研究科, 教授 (70260578)
• Research Collaborator: SASAI Hideo ; AGO Yasuhiko ; OTSUKA Hiroki ; MATSUMOTO Hideki ; MAKAMA Mina
• Project Period (FY): 2016-04-01 – 2019-03-31
• Keywords: 遺伝子 / 遺伝学 / 先天代謝異常症 / ケトン体 / 発現実験

Outline of Final Research Achievements

I have been studying inborn errors of ketone body metabolism. In this study, we tried to establish expression systems of wild-type and mutant cDNAs for HMGCS2 deficiency and HMGCL deficiency of which patients have already been identified in Japan, MCT1 deficiency of which patients have not been identified yet in Japan, and 3HBD deficiency of which patients have never identified in the world. In case of HMGCS2 expression, we failed to get enough expression level for characterization, but we then successfully showed that 4 HMGCS2 mutants reduced enzyme activities significantly using bacterial expression system. In case of 3HBD expression, we established expression system using fetal fibroblast cell line from 3hBDH knock-out mice. We are now trying to establish knock-out cell lines of MCT1 or HMGCL from wild-type fetal fibroblasts.

Free Research Field

小児科学

Academic Significance and Societal Importance of the Research Achievements

HMG-CoA合成酵素欠損症の変異タンパクの発現系の確立をできたことで、日本の症例の確定診断ができ、日本に本症が存在することを論文として報告が可能となった。また日本で症例が同定されていないMCT１欠損症、世界でまだ欠損症の報告がない３HBD欠損症における変異タンパクの評価をできるようにしておくことは今後の迅速な確定診断において重要である。