ICOSL Signaling in Macrophages Promotes Anti-Tumor Immunity

巨噬细胞中的 ICOSL 信号传导促进抗肿瘤免疫

• 批准号: 10251047
• 负责人: Emma Kurz
• 金额: $ 4.45万
• 依托单位: NEW YORK UNIVERSITY SCHOOL OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-09-01 至 2022-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10251047
• 关键词: Acceleration; Adaptor Signaling Protein; Address; Adoptive Transfer; Alanine; Antibodies; Antigen Presentation; Antigen-Presenting Cells; Automobile Driving; Back; Binding; Biological Assay; Biology; Bone Marrow; CD4 Positive T Lymphocytes; CD8B1 gene; Cells; Cytoplasmic Tail; Cytotoxic T-Lymphocytes; Disease; Disease Progression; Family; Flow Cytometry; Genetic; Growth; Human; Immune; Immune response; Immunosuppression; Immunotherapeutic agent; In Vitro; Incidence; Inflammatory; Knowledge; Ligands; Literature; Long-Term Survivors; Malignant Neoplasms; Malignant neoplasm of pancreas; Maps; Mediating; Molecular; Mus; Pancreatic Ductal Adenocarcinoma; Pathway interactions; Pharmacology; Phenotype; Play; Proteins; Regulation; Regulatory T-Lymphocyte; Role; STAT1 gene; Scanning; Signal Pathway; Signal Transduction; Site-Directed Mutagenesis; Surface; Survival Rate; T cell differentiation; T cell response; T-Lymphocyte; Testing; Th2 Cells; Tumor Immunity; Tumor-associated macrophages; Tumor-infiltrating immune cells; Up-Regulation; Work; advanced disease; base; combat; cytotoxic; immunogenic; in vivo; inhibitor/antagonist; macrophage; mortality; mouse model; pancreatic ductal adenocarcinoma model; programs; receptor; recruit; single-cell RNA sequencing; transcriptome; tumor; tumor growth; tumor microenvironment; tumor progression

Project Summary/Abstract: Pancreatic ductal adenocarcinoma (PDA) is an aggressive disease for which there are few long-term survivors. The inflammatory tumor microenvironment (TME) is known to influence pancreatic cancer progression by either generating cytotoxic T cell responses, or, more commonly, by inducing tumor-permissive tolerance. Many studies have shown that tumor-associated macrophages, or TAMs, play a vital role in educating T cells toward immunogenic or tolerogenic differentiation in PDA. However, the mechanisms that regulate macrophage phenotype in this disease are not well understood. ICOSL, the cognate ligand for the ICOS co- stimulatory receptor on T cells, is a B7-family 40 kD protein expressed on macrophages. While ICOS signaling has been well-characterized, the concept of ICOSL ‘back-signaling’ into macrophages upon engagement by ICOS has not been well described. My preliminary work indicates that ICOSL is up-regulated on tumor associated macrophages in the context of pancreatic cancer. Genetic deletion or antibody-mediated neutralization of ICOSL on macrophages results in their differentiation toward an alternatively activated M2-like phenotype and acceleration of cancer progression. Conversely, I found that ICOSL engagement or activation in macrophages by treatment with an ICOS Fc leads to a profound immunogenic program in vitro. Mass spectrometric analysis has shown that ICOSL co-precipitates with STAT1 in macrophages, a known driver of M1-like phenotype. However, the specific molecular mechanisms through which ICOSL acts as a regulator of macrophage phenotype and its applicability as a master regulator of anti-tumor immunity in PDA remains unknown. My proposal will attempt to bridge this gap in knowledge through two main specific aims: Aim 1 will seek to determine if ICOSL-mediated tumor protection in PDA is a T-cell dependent phenomenon. Specifically, this aim will explore the effects of ICOSL-activation or neutralization in macrophages on T-cell phenotype; employing flow cytometry, antigen presentation assays, and 10x single cell RNA sequencing of intra-tumoral immune cells from multiple murine models of PDA. Aim 2 will seek to address the specific intracellular signaling mechanisms through which ICOSL regulates macrophage polarization. By employing site directed mutagenesis to map the interaction surface that mediates ICOSL binding to STAT1, this aim will uncover a heretofore unknown signaling pathway that drives immunogenic macrophage programming involving the cytoplasmic tail of ICOSL.

项目摘要/摘要： 胰腺导管腺癌（PDA）是一种侵略性疾病，很少有长期生存。 已知炎症性肿瘤微环境（TME）会影响胰腺癌的进展 通过诱导的肿瘤耐受性产生细胞毒性T细胞反应，或者更常见。 许多研究表明，肿瘤相关的巨噬细胞或TAM在教育T细胞中起着至关重要的作用 朝PDA中的免疫原性或耐受性分化。但是，调节的机制 该疾病中的巨噬细胞表型尚不清楚。 ICOSL，ICOS共同的同源配体 T细胞上的刺激受体是在巨噬细胞上表达的B7家庭40 kD蛋白。而ICOS信号传导 经过良好的特征，ICOSL在互动时“背信号”构成巨噬细胞 ICO尚未得到很好的描述。我的初步工作表明ICOSL在肿瘤上上调 在胰腺癌的背景下相关的巨噬细胞。遗传缺失或抗体介导的 ICOSL对巨噬细胞的中和导致其分化为替代激活的M2样 癌症进展的表型和加速度。相反，我发现ICOSL参与或激活 在巨噬细胞中，通过ICOS FC治疗，在体外导致了一个深刻的免疫原性程序。大量的 光谱分析表明，ICOSL在巨噬细胞中与STAT1共沉淀，巨噬细胞是已知的驱动因素 M1样表型。但是，ICOSL充当调节剂的特定分子机制 巨噬细胞表型及其作为PDA抗肿瘤免疫的主要调节剂的适用性 未知。 我的建议将尝试通过两个主要目的在知识中弥合这一差距：AIM 1将寻求 确定PDA中ICOSL介导的肿瘤保护是否是T细胞依赖性现象。具体来说，这个目标 将探讨巨噬细胞中ICOSL激活或神经化对T细胞表型的影响；雇用 流式细胞仪，抗原表现测定和10倍单细胞内部免疫细胞的单细胞RNA测序 来自PDA的多个鼠模型。 AIM 2将寻求解决特定的细胞内信号传导机制 ICOSL调节巨噬细胞极化。通过采用定向诱变来绘制 介导ICOSL结合到STAT1的相互作用表面，该目标将发现迄今未知 信号通路驱动涉及ICOSL细胞质尾部的免疫原性巨噬细胞编程。