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

项目概要/摘要： 在临床前模型中，在慢性应激期间由交感神经释放的去甲肾上腺素已经被证实是一种神经刺激。 证明通过激活β2- 肾上腺素能受体（β2AR）在包括免疫细胞在内的各种细胞上的表达。这一系列事件可能是 这对正在经历增加的压力水平的癌症患者的治疗结果有害。先天 淋巴样细胞（ILC），特别是II型ILC（ILC 2），已被证明是一种小而关键的细胞 TME内的人口。然而，关于调节ILC可塑性的机制和 在TME中发挥作用。尽管ILC 2表达高水平的β2AR，但压力如何影响ILC 2在细胞内的活性， TME尚不清楚。在新的初步数据中，我们观察到ILC 2增加与 在缺乏β2AR的敲除小鼠中减少肿瘤体积。此外，我们的数据表明ILC 2的转变 通过单细胞RNA测序，β 2 AR信号转导丧失后，抗肿瘤表型的可塑性。因此，我们认为， 我们假设慢性应激激活的β2AR信号驱动ILC 2的免疫抑制功能， 抑制肿瘤微环境内的抗肿瘤免疫应答。我们将审问这个角色 β 2 AR信号在ILC的发育和可塑性中起着变阻器的作用 在这里提出。在目的1中，我们将确定β2AR信号对普通淋巴祖细胞（CLP）的影响。 分化为ILC 2和ILC 2可塑性。利用野生型和β2AR-/-CLP和ILC 2的体外培养物 在β-AR激动剂治疗的情况下，我们将分析由于β2AR信号传导引起的ILC亚群比率的变化。我们将 还阐明了这些变化发生的分子机制，使用靶向和高通量 方法论。在目标2中，我们将确定β2AR信号传导对ILC 2介导的肿瘤进展的影响。 使用IL-5creβ2-ARfl/fl条件性敲除小鼠。结果表明，小鼠TME和肿瘤生长的变化 将使用光谱流式细胞术分析ILC 2中的条件性β2AR敲除。总的来说，该项目将利用 体外和体内模型以及其他最先进的技术，以了解慢性应激如何通过β2- AR信号传导阻碍肿瘤微环境中有效的抗肿瘤免疫应答。

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.