The immune regulation of macrophage antibody dependent cellular phagocytosis

项目摘要 治疗性抗体对靶细胞的杀伤正在扩大广泛的 一系列自身免疫性疾病和癌症。来自老鼠模型的越来越多的证据，人性化的老鼠 系统和对人体组织的分析表明，巨噬细胞是治疗的主要效应者 抗体，介导被感染的、恶性的和免疫异常细胞的破坏。Fcγ受体 巨噬细胞表面的FCR结合靶细胞相关的单抗 启动抗体依赖的细胞吞噬作用(ADCP)并杀死目标细胞。患者对以下问题的反应 单抗治疗可以从完全缓解到最小的治疗效果。一种未被发掘的可能性 是ADCP反应的变异性是其依赖于巨噬细胞极化的影响 免疫调节的作用。具体地说，免疫抑制环境改变了巨噬细胞的极化引导 到无效的ADCP。相反，干扰素基因激动剂的刺激物(Stinga)，通过1型发挥作用 干扰素(干扰素-1)可显著增强ADCP，克服免疫抑制。 我们的首要目标是阐明巨噬细胞激活控制FCR的机制。 用系统生物学方法研究体内转录组和全基因组的功能和ADCP CRISPR屏幕。ADCP已确定的基因-功能关系将在新的活体模型和 翻译成人类巨噬细胞和治疗性抗体。我们假设ADCP受 巨噬细胞极化的主轴(M0、M1(干扰素γ/脂多糖)、M(干扰素-1/STIN)、M2(IL4/13)和M(S)) 调控FCRs A：I比的未知基因的基因表达变化 和天生的细胞识别受体。我们的提案有两个创新目标，这两个目标将大大扩展 了解FCR依赖的ADCP的调节。在目标1中，我们将阐明巨噬细胞的基因 有助于区分M1、M(干扰素-1/刺激物)、M2和M(S)的FCR功能和ADCP。这一目标需要 一种新的基于CRISPR的全基因组筛选策略在识别促进和抑制基因方面的优势 原代来源巨噬细胞中的ADCP。AIM 2将描绘支持FCR的巨噬细胞基因调控 体内功能和ADCP。在这里，我们将翻译来自患者数据的结果和来自AIM的CRISPR屏幕 1确定ADCP在体内的调控因子。这两个目标都将集中在临床相关的抗B细胞(利妥昔单抗)和 抗T细胞(CamPath)抗体，并将产生扩展我们对FC依赖的理解的发现 ADCP的杀伤机制。

Project Summary The killing of target cells by therapeutic antibodies is expanding the effective treatment options for a wide range of autoimmune diseases and cancers. Mounting evidence from mouse models, humanized mouse systems and the analysis of human tissues, indicates that macrophages are principal effectors of therapeutic antibodies, mediating the destruction of infected, malignant and immunologically aberrant cells. Fcγ receptors (FcR) on the surface of macrophages bind target-cell associated monoclonal IgG class antibodies (mAbs) to initiate antibody-dependent cellular phagocytosis (ADCP) and killing of the target cell. Patient responses to mAb therapies can vary from complete remission to minimal therapeutic effect. One poorly explored possibility is that the variability of the ADCP response is its dependence on macrophage polarization under the influence of immune modulation. Specifically, immunosuppressive environments alter macrophage polarization leading to ineffective ADCP. Conversely, stimulators of interferon genes agonists (STINGa), acting through type 1 interferons (IFN-1) can dramatically potentiate ADCP and overcome immunosuppression. Our overarching goal is to elucidate the mechanistic pathways by which macrophage activation controls FcR function and ADCP using a systems biology approach across in vivo transcriptomics and whole genome CRISPR screens. Identified gene-function relationships for ADCP will be validated in a novel vivo model and translated to human macrophages and therapeutic antibodies. We hypothesize that ADCP is regulated across major axes of macrophage polarization (M0, M1(IFNγ/LPS), M(IFN-1/STING), M2(IL4/13) and M(S)) by gene-expression changes of yet undefined genes that modulate the A:I ratios of FcRs, their signaling machinery and innate cellular recognition receptors. Our proposal has two innovative aims that will vastly expand understanding of the regulation FcR-dependent ADCP. In Aim 1, we will elucidate the macrophage genes contributing to differential FcR function and ADCP across M1, M(IFN-1/STING), M2 and M(S). This aim takes advantage of a new CRISPR-based whole genome screening strategy to identify genes that promote and inhibit ADCP in primary derived macrophages. Aim 2 will delineate macrophage gene regulation supporting FcR function and ADCP in vivo. Here we will translate findings from patient data and the CRISPR screen from Aim 1 to define regulators of ADCP in vivo. Both aims will focus on clinically relevant anti-B cell (Rituximab) and anti-T cell (CAMPATH) antibodies, and will generate findings that extend our understanding of Fc-dependent killing mechanism of ADCP.