Development of novel genome editing tool for gene therapy and its application to cancer therapy

2022 Fiscal Year Final Research Report

• Project/Area Number: 20K21551
• Research Category: Grant-in-Aid for Challenging Research (Exploratory)
• Allocation Type: Multi-year Fund
• Review Section: Medium-sized Section 50:Oncology and related fields
• Research Institution: Tokai University
• Principal Investigator: Ohtsuka Masato 東海大学, 医学部, 教授 (90372945)
• Co-Investigator(Kenkyū-buntansha): 八幡 崇 東海大学, 医学部, 教授 (10398753)
• Project Period (FY): 2020-07-30 – 2022-03-31
• Keywords: ゲノム編集 / 遺伝子治療 / 白血病

Outline of Final Research Achievements

In this study, we aimed to develop novel therapeutic tools that combine the advantages of molecularly targeted drugs and antibody drugs by applying genome-editing technology. Specifically, we attempted to construct a novel tool (scFv-Cas9) by fusing a single-chain antibody fragment (scFv) with Cas9 protein, and used it together with gRNA targeting leukemia-causing chimeric genes. Here, we tried to i) establish a chimeric gene-specific genome editing system using cultured cells, and ii) develop the scFv-Cas9 tool and analyze it in the mouse. The results suggest that deletion of the chimeric gene eliminate leukemic cells. However, the scFv-Cas9 tool itself has not yet been evaluated because of the difficulty in purifying the scFv-Cas9 fusion protein. We will further improve the purification method and investigate other fusion methods.

Free Research Field

遺伝子工学

Academic Significance and Societal Importance of the Research Achievements

in vivo遺伝子治療成功の鍵は、いかに正確且つ適切にゲノム編集ツールを患部の細胞へデリバリーできるかと、いかに副作用を回避できるかなどにある。今回開発を目指した系が期待通りに動くのであれば、標的細胞の特異性と標的DNAの特異性の２点で、キメラ遺伝子を有する白血病細胞のみに治療効果をもたらすさことができ、正常細胞への副作用が回避できると考えられる。それが本システムの特徴であるが、scFv領域やgRNAは他の抗原やDNA領域を標的としたものに自在に変更可能であることから、幅広い分野の研究にも応用可能である。