Novel method for capturing a minimal amount of pancreatic tumor-derived mutations using digital PCR
| Project/Area Number |
20K16947
|
|---|
| Research Category |
Grant-in-Aid for Early-Career Scientists
|
| Allocation Type | Multi-year Fund |
|---|
| Review Section |
Basic Section 53010:Gastroenterology-related
|
|---|
| Research Institution | Asahikawa Medical College |
|---|
Principal Investigator |
|
|---|
| Project Period (FY) |
2020-04-01 – 2022-03-31
|
| Project Status |
Completed (Fiscal Year 2021)
|
| Budget Amount *help |
¥3,770,000 (Direct Cost: ¥2,900,000、Indirect Cost: ¥870,000)
Fiscal Year 2021: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
Fiscal Year 2020: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
|
|---|
| Keywords | 膵癌 / 低侵襲診断 / デジタルPCR / ドライバー遺伝子 / digital PCR / digital 細胞診 / リキッドバイオプシー / 膵腫瘍 / ゲノム診断 |
|---|
| Outline of Research at the Start |
本研究では、微量核酸の絶対定量に威力を発揮するデジタルPCRを用いて、少量の細胞・組織検体でもドライバー変異を確実に検出することで、腫瘍の存在診断を可能にする検出系を確立する。実地臨床において迅速かつ安価に運用可能なシステムを提供する具体的方法として、「ナノ液滴への細胞・核酸封入」によるシングルセル解析を提案する。特に、消化器癌において高頻度にみられるKRASなどの遺伝子変異を複数同時に検出できれば、形態学に基づく画像・病理診断を補填し、がん診療に大きく貢献できる。
|
| Outline of Final Research Achievements |
Histological examinations performed for the definitive diagnosis of pancreatic cancer are often considered unsuitable due to insufficient sample collection. We aimed to develop a technique to accurately determine the genetic abnormality of cancer cells using a small tissue specimen. We established a Water-burst method to detect driver mutations by "encapsulating nucleic acid in droplets" using a digital PCR platform, which allows absolute quantification of minimal amounts of nucleic acid using residual needle biopsy samples from patients with pancreatic cancer tumors. This method has the advantage of skipping nucleic acid extraction from biological samples and rapidly detecting the driver mutations such as KRAS. Furthermore, adjusting the mixing ratio of specific probes for various KRAS mutations made it possible to identify a particular mutation type.
|
| Academic Significance and Societal Importance of the Research Achievements |
本法の実用化により、検体不良（insufficient material）に起因する再生検を減少させることができ、このことは患者への侵襲低減と医療費削減の双方に貢献できる。超微小検体から確実に癌の存在・質的情報が得られれば、再発や薬剤耐性が疑われる場合に、細径穿刺針を用いて少量の組織サンプルを採取し、仮に病理学的な判定ができずとも、遺伝子異常プロファイルを得ることも可能となる。
|
Report
(3 results)
Research Products
(8 results)
-
-
-
[Journal Article] Time-saving method for directly amplifying and capturing a minimal amount of pancreatic tumor-derived mutations from fine-needle aspirates using digital PCR2020
Author(s)
Ono Y, Hayashi A, Maeda C, Suzuki M, Wada R, Sato H, Kawabata H, Okada T, Goto T, Karasaki H, Mizukami Y, Okumura T
-
Journal Title
Scientific Reports
Volume: 10
Related Report
Peer Reviewed / Open Access
-
-
-
-
-
