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）是一种侵袭性疾病，很少有长期幸存者。 众所周知，炎症性肿瘤微环境（TME）通过以下方式影响胰腺癌的进展： 要么产生细胞毒性 T 细胞反应，或更常见的是，通过诱导肿瘤耐受性。 许多研究表明，肿瘤相关巨噬细胞 (TAM) 在培养 T 细胞方面发挥着至关重要的作用 然而，PDA 的免疫原性或耐受性分化的调节机制。 这种疾病中的巨噬细胞表型尚不清楚，ICOSL 是 ICOS 的同源配体。 T 细胞上的刺激受体，是巨噬细胞上表达的 B7 家族 40 kD 蛋白，同时负责 ICOS 信号传导。 ICOSL 的概念已经得到很好的描述，即在巨噬细胞参与时向巨噬细胞发出“反向信号” 我的初步工作表明 ICOSL 在肿瘤上表达上调。 胰腺癌中的相关巨噬细胞。 ICOSL 对巨噬细胞的中和导致巨噬细胞分化为替代激活的 M2 样细胞 线下的表型和加速癌症进展，我发现 ICOSL 参与或激活。 通过用 ICOS Fc 处理巨噬细胞，可在体外产生深远的免疫原性程序。 光谱分析表明 ICOSL 与巨噬细胞中的 STAT1 共沉淀，STAT1 是巨噬细胞中已知的驱动因素 M1 样表型然而，ICOSL 作为调节剂的具体分子机制。 巨噬细胞表型及其作为 PDA 抗肿瘤免疫主要调节因子的应用 未知。 我的建议将尝试通过两个主要具体目标来弥合这一知识差距： 目标 1 将寻求 确定 ICOSL 介导的 PDA 肿瘤保护是否是 T 细胞依赖性现象。 将探索巨噬细胞中 ICOSL 激活或中和对 T 细胞表型的影响； 流式细胞术、抗原呈递测定和肿瘤内免疫细胞的 10x 单细胞 RNA 测序 目标 2 将寻求解决特定的细胞内信号传导机制。 ICOSL 通过采用定点突变来调节巨噬细胞极化。 介导 ICOSL 与 STAT1 结合的相互作用表面，这一目标将揭示迄今为止未知的 驱动免疫原性巨噬细胞编程的信号通路涉及 ICOSL 的细胞质尾部。