| Project/Area Number |
17K16057
|
|---|
| Research Category |
Grant-in-Aid for Young Scientists (B)
|
| Allocation Type | Multi-year Fund |
|---|
| Research Field |
Respiratory organ internal medicine
|
|---|
| Research Institution | Jichi Medical University |
|---|
Principal Investigator |
|
|---|
| Research Collaborator |
Surina
Kobayashi Kunihiko
|
|---|
| Project Period (FY) |
2017-04-01 – 2019-03-31
|
| Project Status |
Completed (Fiscal Year 2018)
|
| Budget Amount *help |
¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000)
Fiscal Year 2018: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
Fiscal Year 2017: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
|
|---|
| Keywords | 肺がん / RNA / 融合遺伝子 / 免役チェックポイント関連因子 / 免役チェックポイント関連遺伝子 / 免疫チェックポイント関連遺伝子 / 臨床呼吸器学 |
|---|
| Outline of Final Research Achievements |
We isolated RNA from bronchial lavage fluid samples from lung cancer patients of our institute, who agreed to participate in the NEJSG021B study. RNA was reverse-transcribed from the oligo(dT) primers using the SuperScript VILO cDNA synthesis kit. Then we detected the ALK, ROS-1 and RET fusion genes using TaqMan based real time PCR assay. The samples in which fusion genes were detected generated amplification products using AMPure parametric beads, and then detailed typing was performed using the direct sequence method. At first, MET expression and PD-L1 expression were detected and analyzed by real-time PCR, normalized and expressed by expression of the housekeeping gene OAZ1. Finally, we constructed a system that can rapidly detect fusion genes such as ALK, ROS-1, RET and MET amplification and PD-L1 expression using next-generation sequencer.
|
| Academic Significance and Societal Importance of the Research Achievements |
我々は適切な操作を行えば肺がん患者の気管支洗浄液からRNAを安定に採取できることを見出し,ALK,ROS1,RETなどの融合遺伝子,さらにはMET発現増幅，PD-L1発現について高速シークセンサーを用いて解析し，得られた成果を迅速に実臨床に返却するシステムの構築を行った.これは肺癌細胞での遺伝子変異解析,発現解析の可能性を大きく広げるものである.少量の気管支洗浄液からRNAを抽出し,PCRで増幅させるため組織採取は不要であり,患者への苦痛軽減のみならず,肺がんの治療に直結する結果を得るという意義がある.本研究は,RNA に基づく肺癌治療マーカーの実用化への道を開く可能性がある.
|
