Suppression of PHOX2B (+7Ala mutant) expression by antisense nucleic acid

2021 Fiscal Year Final Research Report

• Project/Area Number: 20K16927
• Research Category: Grant-in-Aid for Early-Career Scientists
• Allocation Type: Multi-year Fund
• Review Section: Basic Section 52050:Embryonic medicine and pediatrics-related
• Research Institution: Kobe University
• Principal Investigator: Fukushima Sachiyo 神戸大学, 医学研究科, 医学研究員 (20866032)
• Project Period (FY): 2020-04-01 – 2022-03-31
• Keywords: PHOX2B / CCHS / iPS細胞 / ASO

Outline of Final Research Achievements

(1) ASO screening was performed using mini-genes containing Ex1-Ex3 of PHOX2B from genomic DNA derived from patient samples. Four ASOs that specifically suppress the expression of PHOX2B(+7Ala mutant) mRNA were obtained. (2) Four iPS cell lines were established from patient-derived blood samples and induced to differentiate into PHOX2B-expressing neurons. In the future, the effect of ASO on PHOX2B (+7Ala mutant) mRNA suppression will be evaluated in the differentiated neurons.

Free Research Field

新生児

Academic Significance and Societal Importance of the Research Achievements

本研究では、PHOX2B遺伝子の7アラニン伸長変異mRNAの発現をASOで抑制することでCCHSに対する治療法開発を試みた。ミニ遺伝子を利用して得られたASOはPHOX2B (+7Ala mutant) pre-mRNAに特異的に作用し、スプライシングを抑制することで変異mRNAの産生を抑制することが示唆された。さらに、本研究で樹立したCCHS患者由来iPS細胞は、PHOX2B遺伝子の7アラニン伸長変異を標的とするASOや低分子化合物などのスクリーニングに利用できることからCCHSの新規治療法の開発に役立つ。