Inhibition of Hedgehog Signaling in Gli-1+Adeno CA of the Esoph or GE junction

描述（由申请方提供）：食管腺癌（EAC）是一种主要的全球健康负担，其发病率大幅上升。患者预后通常很差，5年治愈率仍<20%。术前化疗放射为治疗局部EAC提供了最强有力的1级证据，然而，这是一种经验性方法，结局不可预测。总之，目前没有一种局部化EAC的方法是基于对分子生物学的基本理解。我们已经产生了令人信服的数据，支持刺猬（Hh）途径在赋予耐药性治疗的核心作用。我们的数据证明，在EAC细胞中，抑制Hh可以克服对细胞毒素和辐射的抗性。我们还开发了一种经验证的3-生物标志物（声波Hh，NF-κ B和Gli-1）签名，用于预测EAC患者对化疗的病理完全反应（pathCR）。因此，Hh信号传导和NF-κ B通路在介导EAC的抗性中显得非常重要。在这个项目中，我们专注于Hh信号。我们的假设是，在具有表达EAC的局部核Gli-1的患者中，在放化疗期间抑制Hh信号传导将导致e40%的pathCR率。GDC-0449（Hh通路的smo抑制剂）在体外和体内下调核Gli-1，并且在体内具有功效。GDC-0449具有抗肿瘤活性，但无剂量限制性毒性（150、270和540 mg）。在这个项目中，我们提出了一个精心设计的翻译临床试验和其他非临床实验，以揭示EAC耐药的分子机制，尽管Hh信号转导的抑制。我们提出了3个具体目标如下：目标1：在具有局部核Gli-1+ EAC的富集患者中进行GDC-0449（NSC 747691）加术前放化疗的IB/II期试验。答：进行IB期试验，以确定GDC-0449加放化疗的安全性。B：进行II期试验，以估计pathCR率并确定GDC-0449的药效学效应（与历史对照相比）。C：进行3-生物标志物pathCR-预测特征的前瞻性验证。目的2：确定目的1中细胞系和患者中GDC-0449耐药的分子途径。答：确定GDC-0449治疗后细胞信号通路中蛋白质的活化和表达状态的变化，并鉴定GDC-0449耐药的关键分子（细胞系和患者）。B：确定GDC-0449治疗后基因表达谱的变化，并建立GDC应答和抗性的生物标志物。目的3：鉴定GDC-0449/放化疗抗性/反应相关的microRNA特征。答：检测EAC细胞株在生物化放疗前后microRNA谱的变化，寻找与耐药相关的microRNA特征。B：确定化疗/放疗耐药EAC患者标本在生物放化疗前后的microRNA谱变化，并鉴定GDC-0449相关的应答/耐药microRNA特征。

DESCRIPTION (provided by applicant): Esophageal adenocarcinoma (EAC) is a major global health burden and its incidence has risen considerably. The patient outcome is often very poor with the 5-year cure rate remaining <20%. Preoperative chemo radiation provides the strongest Level 1 evidence for treating localized EAC, however, this is an empiric approach with unpredictable outcomes. In summary, none of the current approaches to localized EAC are based on fundamental understanding of molecular biology. We have generated compelling data supporting the central role of hedgehog (Hh) pathway in conferring resistance to therapy. Our data document that in EAC cells, inhibition of Hh can overcome resistance to cytotoxics and radiation. We have also developed a validated 3- biomarker (sonic Hh, NF-kB, and Gli-1) signature for predicting pathologic complete response (pathCR) to chemoradiaiton in EAC patients. Thus Hh signaling and the NF-kB pathway appear very important in mediating resistance in EAC. In this Project we focus on Hh signaling. Our hypothesis is that inhibition of Hh signaling during chemoradiation in patients with localized nuclear Gli-1 expressing EAC would result in a e40% pathCR rate. GDC-0449 (a smo inhibitor of Hh pathway) down regulates nuclear Gli-1 in vitro and in vivo and has efficacy in vivo. GDC-0449 has antitumor activity but no dose-limiting toxicity (150, 270, and 540 mg). In this project, we propose an elaborate translational clinical trial and other non-clinical experiments to uncover molecular mechanisms of EAC resistance despite the inhibition of Hh signaling. We propose 3 Specific Aims as follows: Aim 1: To conduct a phase IB/II trial of GDC-0449 (NSC 747691) plus preoperative chemoradiation in enriched patients with localized nuclear Gli-1+ EAC. A: Conduct a phase IB trial to establish safety of GDC-0449 plus chemoradiation. B: Conduct a phase II trial to estimate the rate of pathCR and establish pharmacodynamic effects of GDC-0449 (compare with historical controls). C: Carry out a prospective validation of 3-biomarker pathCR-predicting signature. Aim 2: To identify the molecular pathways of GDC-0449 drug resistance in cell lines and patients in Aim 1. A: Determine the change in activation and expression status of proteins in cell signaling pathways after GDC-0449 treatment and identify key molecules of GDC-0449 resistance (Cell lines and patients). B: Determine the change of gene expression profiles after GDC-0449 treatment and establish biomarkers for GDC response and resistance. Aim 3: To identify the GDC-0449/chemoradiation resistance/response-related microRNA signature. A: To determine the microRNA profile changes in the EAC cell lines before or after biochemoradiation and find the resistance-related microRNA signature. B: To determine the microRNA profile changes in the chemo/radio resistant EAC patients' specimens before or after biochemoradiation and identify the GDC-0449-related microRNA signature of response/resistance.