Targeting the HGF-MET-TWIST1 pathway to overcome EGFR TKI resistance

表皮生长因子受体（EGFR）突变型（mt）非小细胞肺癌（NSCLC）， 最初对EGFR酪氨酸激酶抑制剂（TKI）具有高应答率，然而，耐药是不可避免的。 HGF-MET通路激活和上皮-间质转化（EMT）转录因子（TF）诱导的 间充质表型是EGFR TKI抗性的常见机制。我们已经确定了 肝细胞生长因子（HGF）-MET-TWIST 1轴作为一种新的靶向信号传导轴，可能解释 对EGFR TKI（包括奥希替尼）的新发和获得性耐药。我们公布的数据显示， EMT-TF，TWIST 1是EGFR mt肿瘤发生所必需的，可在体外和体内介导EGFR TKI耐药。 体内通过抑制细胞凋亡。利用我们一流的产品，从基因或生物学角度瞄准TWIST 1 TWIST 1抑制剂去氢骆驼蓬碱使EGFR mt NSCLC对EGFR TKI重新敏感。我们的初步数据表明， HGF增加TWIST 1表达，TWIST抑制可逆转HGF-MET介导的EGFR TKI 体外抗性。因此，TWIST 1可能是连接这些路径的关键目标节点。 中心假设：HGF-MET介导的TWIST 1诱导导致细胞凋亡和EGFR抑制 EGFR mt NSCLC中的TKI耐药，可通过TWIST 1抑制克服。我们将检验这一假设 具体目标如下： 目的1：确定EGFR中HGF依赖性TWIST 1诱导的机制和临床意义 突变型NSCLC。假设：HGF通过磷酸化导致TWIST 1表达和活性增加 通过ERK和/或AKT（Aim 1a）。我们进一步假设HGF-MET激活与增加的 在EGFR TKI初治和获得性耐药背景下，TWIST 1表达和对EGFR TKI的不良应答 患者（目标1b） 目的2：阐明HGF-TWIST 1介导EGFR TKI耐药的机制。假设：TWIST 1 在新发和获得性耐药中介导HGF-MET依赖性EGFR TKI耐药， 体外（aim 2a）和体内（aim 2b-c）细胞凋亡抑制。 目的3：评价TWIST 1抑制剂去氢骆驼蓬碱克服HGF-MET诱导的EGFR TKI的疗效 阻力假设：去氢骆驼蓬碱联合奥希替尼靶向TWIST 1将克服HGF- 在体外和体内获得性耐药环境中，诱导耐药以及MET驱动耐药。 转化影响：在奥希替尼治疗进展后，没有FDA批准的靶向治疗。我们 独特地定位于鉴定TWIST 1作为HGF-MET依赖性EGFR TKI抗性的介导物， 治疗EGFR TKI耐药的新治疗靶点。

Epidermal Growth Factor Receptor (EGFR) mutant (mt) non-small cell lung cancer (NSCLC), initially has a high response rate to EGFR tyrosine kinase inhibitors (TKIs), however, resistance is inevitable. HGF-MET pathway activation and an epithelial-mesenchymal transition (EMT) transcription factor (TF) induced mesenchymal phenotype are commonly observed mechanisms of EGFR TKI resistance. We have identified the hepatocyte growth factor (HGF)-MET-TWIST1 axis as a novel targetable signaling axis that may account for both de novo and acquired resistance to EGFR TKIs including osimertinib. Our published data showed that the EMT-TF, TWIST1 is required for EGFR mt tumorigenesis and can mediated EGFR TKI resistance in vitro and in vivo through suppression of apoptosis. Targeting TWIST1, genetically or pharmacologically with our first-in-class TWIST1 inhibitor, harmine resensitized EGFR mt NSCLC to EGFR TKIs. Our preliminary data demonstrate that HGF increases TWIST1 expression and that TWIST inhibition can reverse HGF-MET mediated EGFR TKI resistance in vitro. Therefore, TWIST1 maybe the critical targetable node that connects these pathways. Central hypothesis: HGF-MET mediated induction of TWIST1 leads to the suppression of apoptosis and EGFR TKI resistance in EGFR mt NSCLC, which can be overcome with TWIST1 inhibition. We will test this hypothesis in the following Specific Aims: Aim 1: Determine the mechanism and clinical significance of HGF-dependent TWIST1 induction in EGFR mutant NSCLC. Hypothesis: HGF leads to increased TWIST1 expression and activity through phosphorylation by ERK and/or AKT (Aim 1a). We further hypothesize that HGF-MET activation correlates with increased TWIST1 expression and poor response to EGFR TKIs in the EGFR TKI de novo and acquired resistance setting in patients (Aim 1b) Aim 2: Elucidate the mechanism of HGF-TWIST1-mediated EGFR TKI resistance. Hypothesis: TWIST1 mediates HGF-MET dependent EGFR TKI resistance in both de novo and acquired resistance through suppression of apoptosis in vitro (aim 2a) and in vivo (aim 2b-c). Aim 3: Evaluate the efficacy of the TWIST1 inhibitor, harmine to overcome HGF-MET induced EGFR TKI resistance. Hypothesis: Targeting TWIST1 with harmine in combination with osimertinib will overcome HGF- induced resistance as well as MET driven resistance in the acquired resistance setting in vitro and in vivo. Translational Impact: There are no FDA approved targeted therapies after progression on osimertinib. We are uniquely positioned to identify TWIST1 as a mediator of HGF-MET-dependent EGFR TKI resistance and a novel therapeutic target for the treatment of EGFR TKI resistance.