2021 Fiscal Year Final Research Report
Comprehensive evaluation of resistance mechanism of the CDK4/6 inhibitor by CRISPR/Cas9 gene editing in ER-positive breast cancer
Project/Area Number |
20K17592
|
Research Category |
Grant-in-Aid for Early-Career Scientists
|
Allocation Type | Multi-year Fund |
Review Section |
Basic Section 55010:General surgery and pediatric surgery-related
|
Research Institution | Keio University |
Principal Investigator |
NAGAYAMA Aiko 慶應義塾大学, 医学部(信濃町), 助教 (00573396)
|
Project Period (FY) |
2020-04-01 – 2022-03-31
|
Keywords | 乳癌 / ゲノム編集 / CRISPR/Cas9 / CDK4/6阻害剤 / 細胞周期 |
Outline of Final Research Achievements |
We utilized MCF7, a model of hormone receptor-positive breast cancer, and transfected Cas9. The stable expression of Cas9 and enzymatic activity were confirmed. The cells were co-cultured with palbociclib, one of approved CDK4/6 inhibitors, for about 2 weeks. The gDNA was extracted from the final product cells, and the barcode associated with each sgRNA was amplified by PCR and analyzed by NGS. The results of the analysis are currently being confirmed, but the preliminary data showed that knockout of Rb induced cell proliferation, which was confirmed in the previous reports. This result suggestied that the assay for identifying candidate genes for the resistance worked.
|
Free Research Field |
乳癌
|
Academic Significance and Societal Importance of the Research Achievements |
本研究はHR陽性乳癌におけるサイクリン依存性キナーゼ(CDK)4/6阻害剤の耐性遺伝子を同定することを目的とする。近年、内分泌療法とCDK4/6阻害剤の併用が無増悪生存期間の延長を示し、標準治療となった。しかし、薬剤耐性の発現と腫瘍再増大が臨床上の問題となっている。そこで、CRISPR/Cas9のゲノム編集技術を用いてWhole-genome knockout screenを行うことで耐性遺伝子を網羅的に探索し、癌細胞におけるCDK4/6阻害剤耐性の原理を明らかにする。本研究の手法を用いることで、新規耐性遺伝子を発見し、薬剤耐性克服の一助となる知見を得ることが可能と考えられる。
|