Autophagy mediated immune stimulation by reovirus in KRAS mutated colorectal cancer

KRAS 突变结直肠癌中呼肠孤病毒自噬介导的免疫刺激

• 批准号: 10359958
• 负责人: Radhashree Maitra
• 金额: $ 43.4万
• 依托单位: YESHIVA UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-01-14 至 2024-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10359958
• 关键词: Address; Adopted; Area; Autophagocytosis; Autophagosome; Biogenesis; Biological Assay; Biomedical Research; Carbamazepine; Cell Survival; Cells; Characteristics; Chloroquine; Clinic; Clinical; Colorectal Cancer; Colorectal Neoplasms; Development; Disease; Double-Stranded RNA; Environment; Enzyme-Linked Immunosorbent Assay; Etiology; Event; FDA approved; Flow Cytometry; Genome; Homing; Hour; Immune; Immune checkpoint inhibitor; Immunization; Immunofluorescence Immunologic; Immunotherapy; Interferons; Interleukin-1 alpha; Interleukin-1 beta; Interleukin-12; Interleukin-6; Interleukins; Intervention; KRAS oncogenesis; KRAS2 gene; Knowledge; Malignant Neoplasms; Mediating; Medical; Microscopy; Molecular; Mus; Mutate; Oncolytic; Ownership; Pathway interactions; Patients; Pattern; Phagolysosome; Plasma; Positron-Emission Tomography; Process; Property; Proteins; Recommendation; Recycling; Reovirus; Research; Research Personnel; Research Support; Role; Science; Stains; Students; T-Lymphocyte; TNF gene; Tamoxifen; Therapeutic; Time; Transforming Growth Factor beta; Transgenic Organisms; Translating; Transmission Electron Microscopy; Treatment Efficacy; Treatment Protocols; Tumor Tissue; Western Blotting; Work; anti-PD1 antibodies; cancer type; checkpoint therapy; cohort; colon cancer cell line; colon cancer patients; cytokine; experimental study; fluorodeoxyglucose; immune activation; improved; inhibitor; insight; mouse model; mutant; neoplastic cell; novel; oncolysis; post intervention; programmed cell death protein 1; programs; response; success; targeted treatment; transmission process; treatment strategy; tumor; tumor microenvironment; undergraduate student

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.

项目概要 癌症免疫疗法取得的令人瞩目的成功也达到了其局限性，并且免疫疗法的功效失败了。 废弃的 KRAS 突变肿瘤占结直肠癌的 45%。这些患者较少 需要开发有效的替代疗法的治疗选择。晚期呼肠孤病毒具有双 链RNA基因组，在致癌的KRAS转化的结直肠肿瘤中显示出治疗功效。 尽管之前研究过其溶瘤特性，但呼肠孤病毒因其免疫功能而越来越受到重视 刺激特性。然而，它参与免疫刺激，恰恰利用了两个密切相关的 免疫和自噬调节途径在很大程度上仍未被探索。初步结果表明 呼肠孤病毒优先利用免疫废弃突变体 KRAS 肿瘤微环境来成功 支持其增殖并破坏肿瘤细胞。此外，呼肠孤病毒还诱导 KRAS 突变微环境中的自噬。知识差距：理解机制至关重要 呼肠孤病毒采用促进突变型 KRAS 驱动的免疫废弃的免疫富集 结直肠癌的微环境。自噬途径在加强该过程中的贡献是 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法成功转化为临床： 增强自噬途径的诱导。我们建议研究呼肠孤病毒介导的模式 自噬机制的诱导并确定分子事件如何转化以改善 KRAS 突变结直肠癌的免疫刺激特征。在这种情况下，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 突变结直肠癌）小鼠模型以及成熟的同基因小鼠 结直肠癌模型从机制上探索两条通路之间的串扰过程。 结果将提高对 KRAS 突变免疫废弃冷肿瘤动态的理解 促进呼肠孤病毒作为免疫刺激剂 影响：该项目将产生双重影响：提供 丰富了生物医学科学相关的研究环境，为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空白。了解 自噬和免疫途径之间的串扰机制，随后得到研究支持 推荐治疗诱导自噬以改善 KRAS 的免疫治疗反应 突变的结直肠癌将具有典型的临床意义。在这一追求中，我们将实现 AIM1：展示 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。 AIM2：检查自噬诱导是否促进呼肠孤病毒介导的免疫刺激 KRAS 突变转基因和同基因小鼠模型中的冷结直肠肿瘤。