Determination of the molecular mechanisms underlying heart development based on genome-wide analysis for congenital heart disease

2018 Fiscal Year Final Research Report

• Project/Area Number: 16K10059
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Pediatrics
• Research Institution: The University of Tokyo
• Principal Investigator: Inuzuka Ryo 東京大学, 医学部附属病院, 講師 (00597560)
• Co-Investigator(Kenkyū-buntansha): 平田 陽一郎 東京大学, 医学部附属病院, 講師 (40447397)
• Research Collaborator: Nakagama Yu ; Nakanishi Toshio ; Takeuchi Jun
• Project Period (FY): 2016-04-01 – 2019-03-31
• Keywords: 先天性心疾患 / RAS/MAPK症候群関連心臓病 / 心臓発生 / ゼブラフィッシュ

Outline of Final Research Achievements

Based on genome-wide analysis and bioinformatic analysis, LZTR1 was selected as a candidate gene for a pedigree with conotruncal anomaly and hypertrophic cardiomyopathy. We performed in-vivo functional analysis for LZTR1 based on zebra fish model using the CRISPR-Cas9 system. A chimeric F0 mutant harboring a 7bp deletion germline allele was identified and backcrossed to wild-type fish to obtain F1 heterozygotes (lztr1del/+). F1 heterozygotes were crossed to generate F2 homozygous mutants (lztr1del/del), which recapitulated hypertrophic cardiomyopathy. Further, immunohistochemical analysis using anti-phospho-ERK antibody indicated active RAS/MAPK signaling, the deleterious signature of NS-associated hypertrophic cardiomyopathy.

Free Research Field

小児循環器

Academic Significance and Societal Importance of the Research Achievements

本研究では、RAS/MAPK症候群関連心臓病におけるLZTR1の重要性が明らかになった。本研究で作出された、疾患モデルゼブラフィッシュは今後RAS/MAPK症候群関連心臓病の病態生理の解明に役立つことが期待される。特に、ゼブラフィッシュから得られる単離心筋細胞は、哺乳動物のそれと異なり、中長期培養、薬剤刺激、RNA阻害実験に適しており、RAS/MAPK症候群関連心臓病の細胞レベルの病態生理を解析する上で、最適なプラットフォームを提供してくれると考えられる。