Investigating the Role of Radiation-induced Cell Death in Innate Lymphoid Cell Activation within Unique Tumor Microenvironments

项目总结： 电离辐射作为“原位”疫苗的作用是基于辐射诱导肿瘤释放的能力 激活抗原提呈细胞以激活CD8+T细胞的抗原。这就是“非局域效应”在 对一个肿瘤的放射可以根除一个远处的肿瘤。然而，临床报告中的“非镜像性” 效应“是罕见的，可能是因为辐射可以促进免疫抑制细胞的产生。而当 辐射用于治疗已经有一个多世纪了，人们仍然很少了解哪些类型的细胞死亡 是由不同类型的辐射方案诱导的，以及这些细胞死亡机制如何影响 炎症环境与抗肿瘤免疫。有趣的是，虽然T细胞对凋亡非常敏感，但 驻留在组织中的固有淋巴样细胞(ILC)具有辐射抗性。组1、组2和组3 ILC由特定的 IL-1细胞因子家族的成员，它们是组织损伤的“警报器”。然而，准确的 细胞死亡在IL-1细胞因子释放和下游免疫反应中的作用一直不是很好 而ILC从未与辐射的免疫反应有关。因此，建议的 实验将通过检查放射治疗诱导的细胞死亡释放机制来解决关键的缺口 肿瘤微环境中特定的IL-1家族细胞因子激活各种ILC亚群(目标1)。K99 指导性实验将涉及患者衍生的有机物质、细胞死亡分析和新颖的 原位小鼠肿瘤模型，以及各种细胞因子的基因敲除小鼠。目标1的结果将导致 随后的R00独立期研究，以确定辐射诱导的免疫反应 调节抗肿瘤免疫(AIM 2)和放射性肺炎和纤维化(AIM)之间的平衡 3)。Aim 3将在我的博士工作的基础上生成IL-22-IRES-BFP报告小鼠来研究这一作用 ILC3诱导的细胞因子在放射性纤维化中的作用总的来说，这项研究将产生前所未有的洞察力 辐射诱导的细胞死亡机制和特定的细胞因子如何引导下游免疫反应 和临床结果。在R99阶段获得的技术培训和职业发展将有所帮助 我启动了我的独立研究计划，并透露了其他问题，以供未来调查。结果 将作为翻译研究的初步数据，以调查两者之间是否存在相关性 辐射诱导的细胞死亡、IL-1细胞因子家族成员的释放、免疫细胞激活和临床 结果。鉴于我在免疫学和放射肿瘤学方面的全面培训和经验，我 独特的定位，利用这两个技能集来解决这些领域和 对放射治疗的疗效机制有基本的了解。

PROJECT SUMMARY: The role of ionizing radiation as an “in situ” vaccine is based on radiation’s ability to induce the release of tumor antigens which activate antigen-presenting cells to prime CD8+ T cells. This mediates the “abscopal effect” in which radiation to one tumor leads to eradication of a distant tumor. However, clinical reports of the “abscopal effect” are rare, possibly because radiation can promote the generation of immunosuppressive cells. While radiation has been used therapeutically for over a century, it is still poorly understood which types of cell death are induced by different types of radiation regimens and how these mechanisms of cell death effect the inflammatory milieu and anti-tumor immunity. Interestingly, while T cells are very sensitive to apoptosis, but tissue-resident innate lymphoid cells (ILC) are radioresistant. Group 1, 2 and 3 ILCs are activated by specific members of the IL-1 family of cytokines which serve as “alarmins” of tissue damage. However, the precise contribution of cell death to IL-1 cytokine release and downstream immune responses has not been well characterized, and ILC have never been implicated in the immune response to radiation. Therefore, the proposed experiments will address critical gaps by examining how radiotherapy-induced mechanisms of cell death release specific IL-1 family cytokines in the tumor micorenvironment to activate various ILC subsets (Aim 1). The K99 mentored experiments will involve the integration of patient derived organoids, cell death assays, and novel orthotopic murine tumor models, as well as knockout mice for various cytokines. Results from Aim 1 will lead to subsequent R00 independent phase investigations to determine how radiation-induced immune responses regulate the balance between anti-tumor immunity (Aim 2) and radiation-induced pneumonitis and fibrosis (Aim 3). Aim 3 will build on my doctoral work generating the IL-22-IRES-BFP reporter mouse to study the role of this ILC3-induced cytokine in radiation-induced fibrosis. Collectively, the research will yield unprecedented insight into how radiation-induced cell death mechanisms and specific cytokines direct downstream immune responses and clinical outcomes. The technical training and career development obtained during the R99 phase will help me to launch my independent research program and reveal additional questions for future investigation. Results from this work will serve as preliminary data for a translational study to investigate if there is a correlation between radiation-induced cell death, release of IL-1 cytokine family members, immune cell activation, and clinical outcomes. Given my comprehensive training and experience in both immunology and radiation oncology, I am uniquely positioned to utilize both skillsets to address important questions at the intersection of these fields and gain fundamental understanding of the mechanisms that underlie the efficacy of radiation therapy.