Elucidation of local infection mechanism by genome infrastructure construction of respiratory filamentous fungal infection and application to diagnosis and treatment

2018 Fiscal Year Final Research Report

• Project/Area Number: 16K09954
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Infectious disease medicine
• Research Institution: National Institute of Infectious Diseases
• Principal Investigator: Umeyama Takashi 国立感染症研究所, 真菌部, 主任研究官 (20360696)
• Project Period (FY): 2016-04-01 – 2019-03-31
• Keywords: 真菌症 / 糸状菌 / ゲノム編集 / ゲノム解析 / 薬剤耐性

Outline of Final Research Achievements

Whole genome sequencing of multi-drug resistant fungi Scedosporium prolificans and Cunninghamella bertholletiae was performed by next-generation sequencer. We constructed a plasmid for CRISPR/Cas9 genome editing in Aspergillus lentulus and Aspergillus viridinutans. We succeeded in site-directed mutation in Cyp51A of clinical isolates by transformation of ribonucleoprotein consisting of Cas9 protein and in vitro synthesized single-guide RNA. By use of this technique, we verified the contribution of amino acid mutation which might be involved in drug resistance.

Free Research Field

医真菌学

Academic Significance and Societal Importance of the Research Achievements

臨床上重要な病原真菌のゲノム基盤構築により、新たな抗真菌薬の標的探索に繋がるような基礎研究が推進されることが期待できる。本研究によって呼吸器における病原糸状菌の感染成立に必要な分子が発見されれば、真菌症治療法や早期診断法の開発への応用などに新しい展開をもたらすものと期待される。また、ゲノム編集技術を導入したことにより、近年大きな問題になっている抗真菌薬耐性問題に対する原因解明につながる可能性も考えられる。