薬剤耐性における lncRNA を機能化する全ゲノム CRISPRa アプローチ
Assaf C Bester1, Jonathan D Lee1, Alejandro Chavez2
1Cancer Research Institute, Beth Israel Deaconess Cancer Center, Department of Medicine and Pathology, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, MA, USA; Ludwig Center at Harvard, Harvard Medical School, Boston, MA, USA.
Cell
|April 21, 2018
まとめ
この研究では,がん薬剤耐性に影響を与える遺伝子を発見するための新しいCRISPRスクリーニングプラットフォーム (DICaS) が導入されています. AMLにおける化学療法抵抗に寄与するGAS6-AS2 lncRNAを含む新しい細胞サイクル,生存,シグナル伝達遺伝子を特定した.
科学分野:
- ゲノミクス
- 分子生物学
- 癌 研究
背景:
- 化学療法への耐性は 癌による死亡の主な原因です
- サイトラビンは急性骨髄性白血病 (AML) の主要な治療法です.
- 薬に対する感受性に影響を与える遺伝的要因を特定することは,がん治療の改善に不可欠です.
研究 の 目的:
- 癌におけるサイトラビン感受性の遺伝的決定因子を特定するために,包括的な全ゲノムCRISPR活性化スクリーニングプラットフォーム (DICaS) を開発し,適用する.
- 薬剤耐性に関与するタンパク質コーディングと長い非コーディングRNA (lncRNA) の両方の遺伝子を機能的に特徴づける.
- 新しい治療目標とAMLや他のがんにおける抵抗メカニズムを明らかにする.
主な方法:
- デュアル・プロテイン・コーディング・アンド・ノン・コーディング・インテグレーテッド・CRISPR・アクティベーション・スクリーニング (DICaS) プラットフォームの開発
- 760人の全癌細胞系から得られた薬剤遺伝子のデータを用いて,推定耐性遺伝子の初期識別.
- 14701のlncRNA遺伝子の特定のCRISPRアクティベーション (CaLR) 戦略を含む,CRISPRアクティベーションを用いたコード化およびlncRNA遺伝子の全ゲノム機能的特徴付け.
主要な成果:
- DICaSは,細胞サイクル調節,生存/アポトーシス,がんシグナル伝達経路に関連する新しい遺伝子を成功裏に特定しました.
- 特定されたGAS6- AS2 lncRNAの転写活性化により,GAS6/ TAM経路が過度に活性化することが示された.
- この過剰活性化はAMLを含む複数の癌における重要な抵抗メカニズムを表しています.
結論:
- DICaSプラットフォームは,コード化および非コード化遺伝子要素の両方を分析することによって,治療目標を発見するための強力で統合されたアプローチです.
- この発見は,GAS6- AS2などのlncRNAが化学療法抵抗を媒介する役割を強調しています.
- この研究は,サイトラビン抵抗性の遺伝的基礎に関する新しい洞察を提供し,AMLやその他の悪性腫瘍の克服のための潜在的な戦略を提供します.
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