Surgical Studies on the Role of MicroRNAs in Esophageal Cancer

背景：尽管与食管切除术相关的发病率和死亡率显著降低， 食道癌患者的存活率仍然很低。在大多数患者中， 化疗和放疗是有益的。事实上，达到完全病理反应的患者 (pCR)在接受化疗和放疗后，食管切除术往往会经历很大的痛苦。 改善生存。不幸的是，目前达到pCR的患者比例很低。改善 食管癌患者食管切除术后的生存率要求我们确定如何增加 达到pCR的患者数量。在其他恶性肿瘤中，已经发现microRNAs（miRs） 作为有效的诊断、预后和预测生物标志物。此外，根据他们的能力， 作为致癌基因或肿瘤抑制因子，目前正在研究基于miR的治疗方法。 研究了miRs在食管癌发病机制和治疗中的作用还没有被证实。 彻底评估。 目的：本研究的主要目的是根据临床研究确定新的生物标志物和治疗靶点。 食管癌细胞中miR的表达和功能分析。 初步发现：我们的初步研究表明a）miR-214- 3 p和miR-199 a-5 p显著地 与食管上皮细胞相比，在食管癌细胞系中下调，B）这种表达 在最初的人类肿瘤样品中验证了模式，c）已经鉴定了特异性结合相互作用 miR-214- 3 p和miR-199 a-5 p与致癌靶点生存素和丝裂原活化蛋白之间的关系 激酶激酶11（MAP 3 K11），d）miR-214- 3 p和miR-199 a-5 p的异位表达 导致食管癌细胞中存活素和MAP 3 K11的水平显著降低，和e）异位 miR-214- 3 p和miR-199 a-5 p的表达导致重要的功能后果;具体来说， 分别增加对化疗诱导的细胞凋亡的敏感性和减少增殖。基于 这些令人兴奋的观察结果，我们假设miR-214- 3 p和miR-199 a-5 p的下调发生在 在食管癌细胞中经常发生，可用于预后、预测和治疗 目的 方法：为了验证这一假设，我们提出了3个具体目标。(1)为了表征miR- 214- 3 p和miR-199 a-5 p在人食管癌标本中的表达及其与临床的相关性 结果。将从50例患者中获得肿瘤和正常食管上皮的治疗前活检组织。 评估miR-214- 3 p和miR-199 a-5 p的表达及其与预后的相关性。(2)到 确定促进肿瘤发生和发展的miR-214- 3 p和miR-199 a-5 p的新靶点， 食道癌我们将利用人食管癌细胞系来确定新的靶点和功能， 食管癌细胞中的miR-214- 3 p和miR-199 a-5 p。(3)为了确定抗肿瘤功效， 体内miR-214- 3 p和miR-199 a-5 p过表达。人类食管癌细胞系将被改造 以稳定表达选定的miR-214- 3 p或miR-199 a-5 p。这些工程细胞将被植入人体 小鼠它们的生长特性和对化疗的反应将与野生型细胞进行比较。 状态：这是重新提交。 影响：美国食管癌的发病率持续上升，尤其是男性 50岁以上的人根据这些人口统计数据，这种疾病代表了VA的一个重要问题 医疗保健系统。改善食管癌患者的预后将极大地提高 履行其医疗保健使命。

Background: Despite significant reductions in the morbidity and mortality associated with esophagectomy, survival for patients with esophageal cancer remains dismal. In most patients, adjunctive therapies including chemotherapy and radiation are beneficial. In fact, patients who achieve a complete pathologic response (pCR) after treatment with chemotherapy and radiation followed by esophagectomy often experience greatly improved survival. Unfortunately, the percentage of patients who currently achieve a pCR is low. Improving survival for esophageal cancer patients following esophagectomy requires that we ascertain how to increase the number of patients who achieve a pCR. In other malignancies, microRNAs (miRs) have been found to serve as effective diagnostic, prognostic, and predictive biomarkers. In addition, based on their ability to function as oncogenes or tumor suppressors, miR-based therapeutic approaches are currently being investigated. The role of miRs in the pathogenesis and treatment of esophageal cancer has not been thoroughly evaluated. Objectives: The primary objective of this study is to identify new biomarkers and therapeutic targets based an analysis of miR expression and function in esophageal cancer cells. Preliminary Findings: Our preliminary studies indicate that a) miR-214-3p and miR-199a-5p are dramatically downregulated in esophageal cancer cell lines compared to esophageal epithelial cells, b) this expression pattern has been verified in initial human tumor samples, c) specific binding interactions have been identified between miR-214-3p and miR-199a-5p with the oncogenic targets survivin and mitogen activated protein kinase kinase kinase 11 (MAP3K11), respectively, d) ectopic expression of miR-214-3p and miR-199a-5p results in a marked decrease in the levels of survivin and MAP3K11 in esophageal cancer cells, and e) ectopic expression of miR-214-3p and miR-199a-5p results in important functional consequences; specifically, increased sensitivity to chemotherapy-induced apoptosis and decreased proliferation, respectively. Based on these exciting observations, we HYPOTHESIZE that downregulation of miR-214-3p and miR-199a-5p occurs frequently in esophageal cancer cells and can be exploited for prognostic, predictive, and therapeutic purposes. Methods: To test this hypothesis, we propose 3 specific aims. (1) To characterize expression of miR- 214-3p and miR-199a-5p in human esophageal cancer specimens and correlate expression with clinical outcomes. Pretreatment biopsies of both tumor and normal esophageal epithelium will be obtained from 50 patients for evaluation of miR-214-3p and miR-199a-5p expression and correlation with outcomes. (2) To identify novel targets of miR-214-3p and miR-199a-5p that contribute to the development and progression of esophageal cancer. We will utilize human esophageal cancer cell lines to identify new targets and functions of miR-214-3p and miR-199a-5p in esophageal cancer cells. (3) To determine the anti-tumor efficacy of miR-214-3p and miR-199a-5p overexpression in vivo. Human esophageal cancer cell lines will be engineered to stably express selected miR-214-3p or miR-199a-5p. These engineered cells will then be implanted in nude mice. Their growth characteristics and response to chemotherapy will be compared to wild-type cells. Status: This is a resubmission. Impact: The incidence of esophageal cancer continues to increase in the Untied Sates, especially in males over the age of 50. Based on these demographics, this disease represents a significant problem in the VA healthcare system. Improving outcomes for esophageal cancer patients will greatly enhance the ability of the VA to fulfill its healthcare mission.