Role of CCR6-CCL20 in biology and chemotherapeutic response in colon cancer

摘要：结肠癌(Coca)仍然是癌症相关死亡的第三大原因，因为 目前提供的治疗早期和晚期Coca的化疗药物未能达到最佳的临床疗效， 患者往往会产生耐药性，并遭受严重的毒性。因此，一个更有效的治疗方案 较低的毒性是改善Coca临床疗效的关键。为了达到这一目标，我们建议 趋化因子受体CCR6及其配体CCL20在细胞和分子中的作用 目前提供的化疗药物(5FU和奥沙利铂)疗效不佳的机制和 开发更有效、毒性更低的治疗方法。拟议的调查是基于我们发布的和 初步数据显示(I)CCR6在Coca细胞和组织中的高表达；(Ii)CCR6的阳性表达 与疾病分期相关；(Iii)CCR6/CCL20具有生物活性，促进肿瘤的迁移和侵袭 Coca，(Iv)CCR6/CCL20激活β-连环蛋白、蜗牛、波形蛋白、α-SMA并下调E-钙粘蛋白的表达 这一轴在维持已知不太敏感的间充质表型中的意义 化疗。在此基础上，我们假设CCR6-CCL20轴支持细胞和分子 重新编程以逃避化疗和开发针对CCR6/CCL20的治疗药物将会有所改善 化疗效果。这一假设将通过以下目标进行检验。目标1：界定CCR6-- 化疗反应差的分子机制(S)。在这个目标下， 利用基因组学和蛋白质组学的方法，我们将确定CCR6/CCL20轴对 癌细胞分子绕道促进细胞和分子表型，帮助省略细胞毒效应 传统的化疗药物。此外，我们将确定阻塞CCR6/CCL20轴是否会有所改善 亚最佳剂量的5-FU和奥沙利铂的细胞毒性。此外，我们将利用临床样本，建立 CCR6/CCL20对化疗疗效的影响目的2：确定CCR6抑制对血管内皮细胞生长的影响 体内化疗反应。利用异种移植和同基因小鼠模型，这一目标将确定 CCR6/CCL20轴抑制对体内5-FU和奥沙利铂疗效的影响同源模型 将使我们能够确定CCR6/CCL20抑制对全身和肿瘤免疫的影响 5-氟尿嘧啶和奥沙利铂。肿瘤和血清的终点分析将决定细胞和分子的变化 提高化疗疗效和器官毒性的机制。完成这一目标将 为使用CCR6/CCL20定向治疗作为一种新的治疗方式提供理论依据。这项建议 研究是新的，因为CCR6/CCL20对细胞和分子重新编程的影响导致了不良反应 5-FU和奥沙利铂的反应在体外和体内都未进行测试。从这项研究中获得的信息将是 在设计一种毒性最小的高效治疗组合方面具有重要意义。这是预料中的 治疗方案将改善Coca的临床结果和患者的生活质量。

Abstract: Colon cancer (CoCa) remains the third leading cause of cancer-related deaths because chemotherapeutics currently offered to treat early and advanced CoCa fail to achieve optimal clinical response, and patients often develop resistance and suffer severe toxicity. Therefore, a more efficacious treatment option with lower toxicity is crucial to improve CoCa clinical outcomes. To achieve this objective, we propose investigating the role of CC chemokine receptor 6 (CCR6) and its ligand CCL20 on cellular and molecular mechanisms contributing to the poor outcome of currently offered chemotherapeutics (5FU and Oxaliplatin) and develop more effective and less toxic treatment. The proposed investigation is based on our published and preliminary data showing (i) higher CCR6 expression in CoCa cells and tissues; (ii) CCR6 expression positively correlates with disease stage; (iii) CCR6/CCL20 is biologically active and promotes migration and invasion of CoCa, (iv) CCR6/CCL20 activates β-catenin, Snail, vimentin, α-SMA and down regulates E-cadherin indicating the significance of this axis in maintaining mesenchymal phenotype known to be less responsive to chemotherapy. Based on this, we hypothesize that the CCR6-CCL20 axis supports cellular and molecular reprogramming to escape chemotherapeutics and developing therapeutics directed to CCR6/CCL20 will improve chemotherapeutic efficacy. The hypothesis will be tested using the following aims. Aim 1: Define the CCR6- mediated molecular mechanism(s) associated with the poor chemotherapeutic response. Under this aim, using genomics and proteomics approach, we will determine the impact of the CCR6/CCL20 axis on the molecular detour cancer cells take to promote cellular and molecular phenotypes that help omit cytotoxic effects of conventional chemotherapeutics. Additionally, we will ascertain if blocking CCR6/CCL20 axis will improve cytotoxicity of 5-FU and Oxaliplatin at a suboptimal dose. Further, using clinical samples, we will establish the impact of CCR6/CCL20 on chemotherapeutic response. Aim 2: Determine the effect of CCR6 inhibition on the chemotherapeutic response in vivo. Using xenograft and syngenic murine model, this aim will determine the impact of CCR6/CCL20 axis inhibition on the efficacy of 5-FU and Oxaliplatin in vivo. The syngenic model will allow us to ascertain the effects of CCR6/CCL20 inhibition on systemic and tumor immunity with or without 5-FU and Oxaliplatin. Endpoint analysis of tumor and serum will determine the alteration in cellular and molecular mechanisms involved in improving the chemotherapeutic efficacy and organ toxicity. Completing this aim will provide the rationale for using CCR6/CCL20 directed therapy as a new treatment modality. This proposed research is novel since the impact of CCR6/CCL20 on cellular and molecular reprogramming contributing to poor 5-FU and Oxaliplatin response is untested both in vitro and in vivo. Information obtained from this study will be significant in designing a highly efficacious treatment combination with minimal toxicity. This anticipated treatment option will improve CoCa clinical outcomes and patient’s quality of life.