Understanding the role of β2AR-signaling on ILC2 differentiation/plasticity and its implications in cancer progression

项目摘要/摘要： 在临床前模型中，交感神经在慢性应激过程中释放的去甲肾上腺素 证明通过激活β-2-促进免疫抑制的肿瘤微环境 肾上腺素能受体(β-2AR)在包括免疫细胞在内的多种细胞上表达。这一系列事件可能是 对承受更大压力的癌症患者的治疗结果不利。先天的 淋巴样细胞(ILCs)，特别是II型ILCs(ILC2s)已被证明是一种小但关键的细胞 TME内的人口。然而，人们对ILC可塑性的调控机制知之甚少。 在TME中运行。尽管ILC2表达高水平的β2AR，但应激如何影响ILC2在 TME尚不清楚。在新的初步数据中，我们观察到ILC2增加和 在缺乏β2AR的基因敲除小鼠中，肿瘤体积减少。此外，我们的数据表明ILC2发生了转变 单细胞核糖核酸测序显示β-2AR信号缺失时抗肿瘤表型的可塑性。因此， 我们假设慢性应激激活的β2AR信号驱动ILC2的免疫抑制功能， 抑制肿瘤微环境内的抗肿瘤免疫反应。我们将审问这个角色 β-2AR信号在ILC的发育过程中起着变阻器的作用，并通过这两个目的可塑性地进入辅助ILC亚群 在这里求婚。在目标1中，我们将确定β2AR信号对共同淋巴系祖细胞(CLP)的影响 分化为ILC2和ILC2可塑性。利用野生型和β2AR-/-CLP和ILC2的体外培养 通过β-AR激动剂治疗，我们将分析β-2AR信号转导引起的ILC亚群比例的变化。我们会 也阐明了这些变化发生的分子机制，通过靶向和高通量 方法论。在目标2中，我们将确定β2AR信号在ILC2介导的肿瘤进展中的影响 采用IL-5creβ2-ARFL/FL条件性基因敲除小鼠。多发性硬化小鼠TME和肿瘤生长的变化 对ILC2中条件性β-2AR基因敲除进行光谱分析。总体而言，该项目将利用 体外和活体模型和其他最先进的技术来了解慢性应激是如何通过β2- AR信号在肿瘤微环境中阻碍了有效的抗肿瘤免疫反应。

Project Summary / Abstract: In preclinical models, norepinephrine released by sympathetic nerves during chronic stress have been demonstrated to promote an immunosuppressive tumor microenvironment (TME) through activation of the β2- adrenergic receptor (β2AR) on various cells including immune cells. This sequence of events could be detrimental to the treatment outcome in cancer patients who are experiencing increased levels of stress. Innate lymphoid cells (ILCs), specifically type II ILCs (ILC2s), have been demonstrated as a small but critical cell population within the TME. However, there is little known about the mechanisms that regulate ILC plasticity and function in the TME. Even though ILC2s express high levels of β2AR, how stress impacts ILC2 activity within the TME is not yet known. In new preliminary data, we have observed a correlation between increased ILC2s and decreased tumor volume in knock out mice lacking the β2AR. Furthermore, our data suggests a shift in ILC2 plasticity toward an anti-tumor phenotype upon loss of β2AR signaling by single-cell RNA sequencing. Therefore, we hypothesize that β2AR signaling activated by chronic stress drives the immunosuppressive function of ILC2s, suppressing the anti-tumor immune response within the tumor microenvironment. We will interrogate the role β2AR signaling plays as a rheostat in ILC development and plasticity into the helper ILC subsets by the two aims proposed here. In Aim 1, we will determine the effect of β2AR signaling on common lymphoid progenitors (CLPs) differentiation into ILC2s and ILC2 plasticity. Utilizing in vitro cultures of wildtype and β2AR-/- CLPs and ILC2s with a β-AR agonist treatment, we will analyze the changes in ILC subset ratios due to β2AR signaling. We will also elucidate the molecular mechanism by which these changes occur using both targeted and high-throughput methodologies. In Aim 2, we will determine the impact of β2AR signaling on ILC2-mediated tumor progression using IL-5creβ2-ARfl/fl conditional knockout mice. The changes in the TME and tumor growth in mice with a conditional β2AR knock out in ILC2 will be analyzed using spectral flow cytometry. Overall, this project will utilize in vitro and in vivo models and other state-of-the-art techniques to understand how chronic stress through β2- AR signaling hampers effective ant-tumor immune response in tumor microenvironment.