Functional analysis of HSD 10 disease

• Project/Area Number: 17K16245
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: Pediatrics
• Research Institution: Gifu University
• Principal Investigator: Sasai Hideo 岐阜大学, 医学部附属病院, 助教 (20509781)
• Project Period (FY): 2017-04-01 – 2019-03-31
• Project Status: Completed (Fiscal Year 2018)
• Budget Amount: ¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000); Fiscal Year 2018: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000); Fiscal Year 2017: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
• Keywords: HSD10病 / 先天代謝異常症 / ケトン体

Outline of Final Research Achievements

HSD10 disease is a rare X-linked recessive disorders caused by a mutation in the HSD17B10 gene. HSD10 is a multifunctional protein which has three functions. Clinical severity of this disorder is various ranging neonatal severe form to atypical form. The HSD17B10 gene was incorporated into pET28a and the vectors with wild-type cDNA or with each mutant cDNA (A154T, A157V, R226Q) were used to produce high-purity recombinant protein in E. coli expression system. The enzyme activity of 2M3HBD in the isoleucine metabolism system was measured spectrophotometrically. The enzyme activity of 2M3HBD in the mutant protein was significantly decreased compared to wild-type. Also, as compared with A154T and A157V, the enzyme activity was lower in R226Q. In this experiment, there is a certain correlation between the decrease in activity of 2M3HBD and the clinical severity in the isoleucine metabolism system. We are planning to study the further functional analysis.

Academic Significance and Societal Importance of the Research Achievements

申請者らはこれまで、ケトン体代謝異常症の遺伝子解析を行ってきた。そして、国内初のHSD10病の幼児例を含む本症3症例を明らかにしており、国内での解析は申請者らが中心であり独創的であるといえる。HSD10病の詳しい病態や長期予後は分かっておらず、本研究の結果は国内外問わず学術的にも臨床的にも大きな意味があるものと考える。

Research Products

• [Journal Article] Recent advances in understanding beta-ketothiolase (mitochondrial acetoacetyl-CoA thiolase, T2) deficiency (2019)
  • Author(s): Fukao T, Sasai H, Aoyama Y, Otsuka H, Ago Y, Matsumoto H, Abdelkreem E.
  • Journal Title: Journal of Human Genetics Volume: 64 Issue: 2 Pages: 99-111
  • DOI: 10.1038/s10038-018-0524-x
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Intronic antisense Alu elements have a negative splicing effect on the inclusion of adjacent downstream exons (2018)
  • Author(s): Nakama M, Otsuka H, Ago Y, Sasai H, Abdelkreem E, Aoyama Y, Fukao T.
  • Journal Title: Gene Volume: 664 Pages: 84-89
  • DOI: 10.1016/j.gene.2018.04.064
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Presentation] 代謝性疾患マススクリーニング診療における遺伝子検査の重要性(意義) AMED深尾班の遺伝子パネルの現状 (2018)
  • Author(s): 笹井 英雄, 深尾 敏幸
  • Organizer: 日本マス・スクリーニング学会学術集会 (第45回)
• [Presentation] Gene panel study for target metabolic diseases in newborn mass screening (2018)
  • Author(s): Sasai H, Ago Y, Otsuka H, Hosokawa J, Fujiki R, Ohara O, Nakajima Y, Ito T, Hara K, Kobayashi M, Tajima G, Sakamoto O, Matsumoto S, Nakamura K, Hamazaki T, Kobayashi H, Hasegawa Y, Fukao T
  • Organizer: Annual symposium of the society for the study of inborn errors of metabolism 2018
  • Int'l Joint Research
• [Presentation] Functional analysis of mutant recombinant HSDl7Bl0 proteins using an E. Coli expression system (2018)
  • Author(s): Sasai H, Ohnishi H, Akagawa S, Akiba K, Hasegawa Y, Kobayashi M, Otsuka H, Aoyama Y, Ago Y, Fukao T
  • Organizer: Annual symposium of the society for the study of inborn errors of metabolism 2018
  • Int'l Joint Research
• [Presentation] Gene panel study for target metabolic diseases of newborn mass screening in Japan (2018)
  • Author(s): 笹井英雄、吾郷耕彦、松本英樹、大塚博樹、細川淳一、藤木亮次、小原收、中島葉子、伊藤哲哉、小林弘典、長谷川有紀、原圭一、小林正久、但馬剛、坂本修、城戸淳、松本志郎、中村公俊、濱崎孝史、深尾敏幸
  • Organizer: 日本先天代謝異常学会総会 (第60回)
• [Presentation] リコンビナントHSD17B10タンパクを用いたHSD10病の病態解析 (2017)
  • Author(s): 笹井英雄、大西秀典、赤川翔平、秋葉和壽、長谷川行洋、小林正久、大塚博樹、青山友佳、深尾敏幸
  • Organizer: 第59回 日本先天代謝異常学会学術集会