Development of a new gallbladder cancer mouse model using CRISPR-Cas9 system to evaluate the effect of personalized therapy

2022 Fiscal Year Final Research Report

• Project/Area Number: 20K22816
• Research Category: Grant-in-Aid for Research Activity Start-up
• Allocation Type: Multi-year Fund
• Review Section: 0901:Oncology and related fields
• Research Institution: Kyushu University
• Principal Investigator: Ideno Noboru 九州大学, 医学研究院, 助教 (90883421)
• Project Period (FY): 2020-09-11 – 2023-03-31
• Keywords: Gallbladdre cancer / CRISPR-Cas9 system

Outline of Final Research Achievements

We developed a transgenic mouse model Krt19-CreERT; LSL-Cas9, specifically expressing Cas9 in biliary epithelium. Based on available public data, we selected KRAS, TP53, SMAD4 as frequent drivers for gallbladder cancer. As potential therapeutic target, CDKN2A was added to 2 drivers, KRAS/TP53, or SMAD4/TP53. We designed guide RNA to induce gain-of-function mutations in these drivers. After confirmation that each guide RNA worked for genome editing in vitro study, guide RNAs for 2 or 3 driver genes were cloned into AAV vector and generated and purified AAV. The completed works for this study were development of CK19-Cas9 mouse model and generation of AAV containing guide RNA for 2 or 3 driver genes. We are starting injection of AAV into gallbladder of Krt19-CreERT; LSL-Cas9 mice once sufficient number of mice is available, and evaluating phenotypic difference according to edited drivers.

Free Research Field

胆膵悪性腫瘍

Academic Significance and Societal Importance of the Research Achievements

胆嚢癌には併存する複数の仮想的driver遺伝子がある．これらが併存した場合の各遺伝子変異の機能が明らかではないため，遺伝子変異を標的とした治療の有効性を検証するためには，同じ組み合わせの仮想的driver遺伝子を持つpre-clinical modelが必要である．申請者が開発中のCRISPR/Cas9 systemによる胆嚢癌遺伝子改変マウスモデルを応用して，ヒト胆嚢癌の遺伝子情報に基づいて癌を自然発生させたモデルを短期間で作成し，分子標的薬の効果を予測することで，それぞれの患者により効率的な治療を提供できる可能性があり，予後不良な胆嚢癌の治療成績向上に大きく寄与すると考えられる．