2021-01-012024-05-13https://scholars.lib.ntu.edu.tw/handle/123456789/652591"非小細胞肺癌(NSCLC)為致死率最高癌症死因之一，第一代上皮生長因子受體 (EGFR) Tyrosine kinase inhibitor (TKI) gefitinib (Iressa®)或erlotinib (Trcevaa®)，可有效治療EGFR activating mutation的NSCLC。然而，使用後約一年會產生acquired resistance，此抗藥性產生通常是由於EGFR產生第二處 T790M 突變。第三代T790M-specific TKI AZD9291雖能克服EGFR T790M的acquired resistance，但仍會產生新的抗藥性。 我們先前的研究發現，gefitinib acquired resistant HCC827細胞(HCC827/IR)及AZD9291 acquired resistant H1975細胞(H1975/AR) 皆具EMT特性，並增加癌幹細胞的產生，是造成EGFR-TKI 抗藥性的原因。HMGR-HDAC雙標靶藥物(JMF compounds)，分別阻斷Src/Hakai/E-cadherin途徑及抑制Zeb1表現而抑制EMT及癌幹細胞特性，進而克服gefitinib與AZD9291抗藥性 (已發表於Oncogene, 2019；由106R39012計畫支持)。 進一步以RNA sequencing和cDNA microarray分析，是否有新的分子參與NSCLC對EGFR-TKI之抗藥性，發現AKT3、轉錄因子FOXC1在兩株抗藥性細胞及long noncoding RNA-LncDC在第三代抗藥性細胞之表現皆增加，因此希望此PPG獲得支持，進一步探討它們對TKI抗藥性、EMT與癌幹細胞特性間之調控關係。此PPG由三個子計畫組成：子計畫一由陳青周教授執行，子計畫二由魏子堂助理教授執行，並由陳青周教授與施金元教授指導，子計畫三由內科部施金元教授執行。 子計畫一：探討AKT3在非小細胞肺癌EGFR-TKI acquired resistance的機制和調控 目標一：探討AKT3在TKI抗藥性細胞中的過度表達，是否與抗藥性、遷移和存活有關。 目標二：探討為何抗藥性細胞處理TKI，AKT3蛋白會進一步增加但mRNA沒有變化。 目標三：探討AKT抑制劑併用TKI是否有加乘抑制抗藥性細胞存活，並確認病患在處理TKI治療前後， AKT3之角色 子計畫二：探討FOXC1在NSCLC對EGFR-TKI acquired resistance的角色 目標一：在TKI敏感性細胞中過度表達FoxC1，探討是否與抗藥性、細胞幹性和EMT有關 目標二：探討過表達FOXC1之H1975細胞，是否會促進肺癌原位小鼠腫瘤生長與轉移，並測試 AZD9291療效 目標三：探討病人以TKI治療前後，FOXC1的表現 子計畫三：探討miR-204/FOXC1在EGFR-TKI acquired resistance的角色 目標一：確認miR-204與FOXC1之相關性 目標二：探討miR-204是否透過調控FOXC1而抑制EGFR TKI抗藥性 目標三：探討病人以TKI治療前後，miR-204與FOXC1之相關性 此整合型計畫將為NSCLC對EGFR-TKI抗藥性機轉帶來新的見解，將基礎研究轉譯至臨床，解決現有臨床存在問題。" "Non-small-cell lung cancer (NSCLC) is an important cause of cancer death worldwide. The discovery of EGFR-TKI such as gefitinib (Iressa®) or erlotinib (Tarceva®) leads to a great success against NSCLC with EGFR activating mutation. However, acquired resistance emerges after a period of treatment which is largely attributed to secondary EGFR T790M mutation. Although T790M-specific TKI osimertinib (Tagrisso®; AZD9291) can overcome the acquired resistance, their effect is hindered by newly developed acquired resistance. We developed the first and third generation of EGFR-TKI acquired resistant NSCLC cells (HCC827/IR and H1975/AR) not exhibiting additional EGFR mutation, and the involvement of EMT and enrichment of cancer stem cell (CSC) in the acquired resistance were revealed. Our dual HMGR-HDAC inhibitor JMF overcame the acquired resistance of gefitinib and AZD9291 by reversing EMT and stemness through interrupting Src/Hakai/E-cadherin pathway and Zeb1 expression, respectively. These results not only reveal a common acquired resistant mechanism of EGFR TKI beyond EGFR mutations per se but also provide a strategy to overcome it (Oncogene 2019. Supported by106R39012). To further elucidate the underlying mechanisms of EGFR-TKI acquired resistance, next generation RNA-seq and cDNA microarrays are performed to identify novel molecules contributing to acquired resistance. Upregulations of AKT3 and FOXC1 in both resistant cells as well as LncDC in H1975/AR cells are identified. Therefore, their roles in TKI resistance, EMT, and stemness will be explored in this PPG consisting of three subprojects: Sub-project 1 by Prof. Ching-Chow Chen, Sub-project 2 by Assistant Prof. Tzu-Tang Wei under the guidance of Prof. Chen and Shih, and Sub-project 3 by Prof. Jin-Yuan Shih at Internal Medicine. Sub-project 1: Exploring the mechanism and regulation of AKT3 in acquired resistance to EGFR-TKI Aim 1: To study whether AKT3 overexpression in TKI-resistant cells is involved in the resistance and migration. Aim 2: To study how AKT3 protein but not mRNA is further increased in resistant cells treated with TKI Aim 3: To investigate whether combination of AKT inhibitors with TKI exerts synergistic inhibition on cell viability in TKI resistant cells Sub-project 2: To investigate the role of FOXC1 in acquired resistance to EGFR-TKI Aim 1: To investigate whether FoxC1 overexpression in TKI-sensitive cells is involved in the resistance, stemness and EMT Aim 2: To examine whether overexpression of FOXC1 in H1975 cells enhances orthotropic tumor formation and metastasis in mice Aim 3: To confirm the role of FOXC1 in patients treated with TKI. Sub-project 3: To study the role of miR-204/FOXC1 in the resistance to EGFR-TKI Aim 1: Confirm the correlation and interaction of miR-204 and FOXC1. Aim 2: To evaluate whether miR-204 reverses TKI resistance through targeting FOXC1 signaling Aim 3: To confirm the association of miR-204 and FOXC1 in patients treated with EGFR TKIs. This integrated work will bring novel insights for NSCLC resistance to EGFR-TKI and translate the basic research to the clinic to tackle the existing clinical problems."AKT3FOXC1LncDCTKI抗藥性上皮細胞間質轉化癌幹性TKI acquired resistanceEMTstemness