Autophagy mediated immune stimulation by reovirus in KRAS mutated colorectal cancer

项目摘要 令人印象深刻的成功的癌症免疫治疗达到了其局限性，并未能在其疗效与免疫 废弃的KRAS突变肿瘤占结直肠癌的45%。这些患者的 需要开发有效替代疗法的治疗选择。晚期呼肠孤病毒，具有双重- 链RNA基因组的基因在致癌KRAS转化的结直肠肿瘤中显示出治疗功效。 尽管先前研究了其溶瘤特性，但呼肠孤病毒因其免疫原性而越来越受到重视。 刺激特性然而，它参与免疫刺激，正是利用两个密切相关的 免疫和自噬调节途径，仍然在很大程度上未被探索。初步结果表明 呼肠孤病毒优先利用免疫遗弃突变KRAS肿瘤微环境， 支持其传播并破坏肿瘤细胞。此外，呼肠孤病毒诱导 KRAS突变微环境中的自噬。知识差距：了解机制至关重要 通过呼肠孤病毒的免疫富集促进突变KRAS驱动的免疫弃养 大肠癌的微环境自噬途径在加重这一过程中的作用是 不清楚了解自噬和免疫途径之间的控制机制至关重要。 项目假设：呼肠孤病毒发挥的双重作用模式可以通过以下方式成功地转化为临床： 增强自噬途径的诱导。我们建议研究呼肠孤病毒介导的 诱导自噬机制，并确定分子事件是如何翻译的，以改善自噬机制。 KRAS突变的结直肠癌的免疫刺激特征。在这方面，我们将利用 KRAS突变型和野生型结直肠癌细胞系以及我们新开发的KPC：APC（他莫昔芬- 可诱导的KRAS突变的结肠直肠癌）小鼠模型沿着良好建立的同基因小鼠 结直肠癌模型，以机械地探索两种途径之间的串扰过程。 结果将提高对KRAS突变的免疫遗弃冷肿瘤的动力学的理解， 促进呼肠孤病毒作为免疫刺激剂的作用：该项目将产生双重影响， 丰富的生物医学科学相关的研究环境，以有抱负的本科生，以及 直接解决KRAS突变结直肠癌患者面临的临床差距。了解 自噬和免疫途径之间的相互作用机制，研究支持 推荐治疗性诱导自噬以改善KRAS的免疫治疗反应性 突变的结肠直肠癌将具有典型的临床意义。在这一追求中，我们将目标1：证明 在呼肠孤病毒处理的KRAS突变体和野生型结直肠中自噬和免疫途径之间的串扰 癌细胞系。AIM 2：检查自噬诱导是否促进呼肠孤病毒介导的免疫刺激， 转基因和同基因小鼠模型中KRAS突变的冷结直肠肿瘤。

Project Summary The impressive success of cancer immune therapy reached its limitations and failed in its efficacy with immune deserted KRAS mutated tumors that constitute 45% of colorectal cancers. These patients have fewer treatment options necessitating the development of effective alternate therapy. Of late reovirus, with a double- stranded RNA genome, showed therapeutic efficacy in an oncogenic KRAS transformed colorectal tumors. Although previously studied for its oncolytic properties reovirus is being increasingly appreciated for its immune stimulation properties. However, its participation in immune stimulation, precisely utilizing two intimately related immune and autophagic modulatory pathways, remains largely unexplored. Preliminary results indicate that reovirus preferentially exploits the immune deserted mutant KRAS tumor microenvironment to successfully support its propagation in conjunction with destruction of the tumor cells. Furthermore, reovirus induces autophagy in KRAS mutated microenvironment. Knowledge gaps: It is crucial to understand the mechanism adopted by reovirus to facilitate the immune enrichment of mutant KRAS driven immune deserted microenvironment in colorectal cancer. The contribution of autophagic pathway in accentuating the process is unclear. Understanding the control mechanism between the autophagy and immune pathways is critical. Project hypothesis: The dual mode of action exerted by reovirus can be successfully translated to the clinic by augmenting the induction of the autophagy pathway. We propose to study the pattern of reovirus mediated induction of the autophagic machinery and determine how the molecular event is translated in improving the immune stimulation characteristics in KRAS mutated colorectal Cancer. In this context we will make use of KRAS mutant and wildtype colorectal cancer cell lines and our newly developed KPC:APC (tamoxifen- inducible KRAS-mutated colorectal cancer) mouse models along with the well-established syngeneic mouse model of colorectal cancer to mechanistically explore the process of crosstalk between the two pathways. Results will improve the understanding of the dynamics of KRAS mutated immune deserted cold tumors in facilitating reovirus to act as an immune stimulator Impact: This project will have dual impact of providing an enriched biomedical science related research environment to the aspiring undergraduate students as well as to directly address the clinical gap faced by the KRAS mutated colorectal cancer patients. Understanding the mechanism of cross talk between the autophagy and immune pathways followed by research supported recommendation of therapeutic induction of autophagy to improve immune therapy responsiveness of KRAS mutated colorectal cancer will be of paradigm clinical implications. In this pursuit we will AIM1: Demonstrate crosstalk between autophagy and immune pathways in reovirus treated KRAS mutant and wildtype colorectal cancer cell lines. AIM2: Examine if autophagy induction promotes reovirus mediated immune stimulation in KRAS mutated cold colorectal tumors in transgenic and syngeneic mouse models.