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为广泛的免疫缺陷扩大了有效的治疗选择。 一系列自身免疫性疾病和癌症。来自小鼠模型的证据，人源化小鼠 系统和人体组织的分析表明，巨噬细胞是治疗性免疫缺陷的主要效应子。 抗体，介导感染的、恶性的和免疫异常的细胞的破坏。Fcγ受体 (FcR)在巨噬细胞的表面上结合靶细胞相关的单克隆IgG类抗体（mAb）， 启动抗体依赖性细胞吞噬作用（ADCP）并杀死靶细胞。患者反应 mAb疗法可以从完全缓解到最小治疗效果变化。一个未被充分研究的可能性 ADCP反应的可变性是其对巨噬细胞极化的依赖性， 免疫调节。具体而言，免疫抑制环境改变巨噬细胞极化， 到无效的ADCP。相反，干扰素基因激动剂（STINGa）的刺激剂，通过1型 干扰素（IFN-1）可显著增强ADCP并克服免疫抑制。 我们的首要目标是阐明巨噬细胞激活控制FcR的机制途径 使用跨体内转录组学和全基因组的系统生物学方法， CRISPR筛选。已鉴定的ADCP基因-功能关系将在新的体内模型中得到验证， 转化为人类巨噬细胞和治疗性抗体。我们假设ADCP是受监管的， 巨噬细胞极化的主轴（M0、M1（IFNγ/LPS）、M（IFN-1/STING）、M2（IL 4/13）和M（S）） 调节FcRs的A：I比率的尚未确定的基因的基因表达变化，其信号机制 和先天细胞识别受体。我们的提案有两个创新目标， 了解FcR依赖性ADCP的调节。在目标1中，我们将阐明巨噬细胞基因 有助于M1、M（IFN-1/STING）、M2和M（S）之间的差异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的优势， 原代衍生巨噬细胞中的ADCP。目的2阐明巨噬细胞支持FcR的基因调控 功能和ADCP。在这里，我们将从患者数据和Aim的CRISPR筛选中翻译结果 1来定义ADCP在体内的调节物。这两个目标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，并将产生的结果，扩大我们的理解Fc依赖性 ADCP的杀伤机理

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.