Visualize gene point mutations from cytological specimens to elucidate the mechanisms that cause cell atypia

2022 Fiscal Year Final Research Report

• Project/Area Number: 20K16198
• Research Category: Grant-in-Aid for Early-Career Scientists
• Allocation Type: Multi-year Fund
• Review Section: Basic Section 49020:Human pathology-related
• Research Institution: Wakayama Medical University
• Principal Investigator: Matsuzaki Ibu 和歌山県立医科大学, 医学部, 助教 (60647428)
• Project Period (FY): 2020-04-01 – 2023-03-31
• Keywords: 細胞診 / 遺伝子変異 / 細胞異型

Outline of Final Research Achievements

The purpose of this study was to develop a method for visualizing gene point mutations in situ using cytological specimens and to analyze morphological changes. Prior to in situ visualization, I examined whether there were differences in cell morphology and detected genes depending on the fixation solutions normally used for cell diagnostic materials.As a result, morphological changes occurred depending on the type of fixative and fixation time, and there was a tendency for artifacts to occur in the mutated genes detected by next generation sequencing.
We examined the detection of point mutations using the BaseScope and mutaFISH methods, suggesting that the presence or absence of mutations causes differences in detection signals in case of using cultured cells.

Free Research Field

細胞診断学

Academic Significance and Societal Importance of the Research Achievements

細胞診は遺伝子解析材料としての有用性が注目されているが、Liquid-based cytology用の固定液の種類および固定時間によって、形態学的変化が生じるのみならず、遺伝子解析で検出される遺伝子変異にアーチファクトが生じる可能性があり、解析や解釈に注意が必要である。
in vivoでの個々の細胞ごとの遺伝子変異検出は、mutaFISH法などの培養細胞を用いての条件検討は終了した。
今後、細胞診材料に応用できれば、治療戦略を立案するためのコンパニオン診断に寄与できることが期待される。また、遺伝子変異を有無により細胞の形態学的変化を見いだせれば、細胞診断の正診率が向上できる可能性がある。