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Deciphering the molecular control of intratumoral dendritic cells

破译瘤内树突状细胞的分子控制

基本信息

批准号：
10559621
负责人：
Brian D Brown
金额：
$ 53.63万
依托单位：
ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-02-01 至 2026-01-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10559621
关键词：
ALDH1A2 gene Address Antigen Presentation Antigens Antitumor Response BIRC2 gene BIRC3 gene CD 200 CD8-Positive T-Lymphocytes Cell Death Cell Maturation Cell Shape Cell physiology Cells Cellular biology Charge Clinical Clinical Trials Clustered Regularly Interspaced Short Palindromic Repeats Cross Presentation Data Dendritic Cells Dendritic cell activation Equilibrium Genes Genetic Transcription Goals Human IL12B gene IL4 gene IL4R gene Immune Immunity Immunophenotyping Immunotherapy Individual Infiltration Inhibition of Apoptosis Interleukin-12 Interleukin-4 Knock-out Lesion Lung Neoplasms Lymphocyte Activation Macrophage Mediating Modeling Molecular Molecular Profiling Mus NF-kappa B NFKB Activation Pathway Nature Non-Small-Cell Lung Carcinoma Outcome Output PD-1 blockade Pathway Analysis Pathway interactions Phenotype Physiology Population Pre-Clinical Model Production Regulator Genes Resolution Role Signal Transduction T cell response T-Lymphocyte TNFRSF5 gene TNFSF4 gene Testing Therapeutic Tumor Antigens Tumor Burden Tumor Expansion Tumor Immunity Tumor Promotion anti-PD-1 cancer cell cancer immunotherapy cancer infiltrating T cells cell motility cytokine draining lymph node effector T cell genomic platform immunoregulation improved in vivo inhibitor interest mouse model pharmacologic pre-clinical prevent programmed cell death ligand 1 programmed cell death protein 1 programs response single-cell RNA sequencing synergism tumor tumor growth ubiquitin ligase uptake

项目摘要

PROJECT SUMMARY Dendritic cells (DC) have a major influence on tumor immunity. DC depletion abrogates tumor immunity and response to PD1 (αPD1) immunotherapy in many preclinical tumor models and DC infiltration in human tumors is a positive correlate of clinical outcome. Unfortunately, we still lack effective strategies for harnessing DC to stimulate tumor immunity and this is in large part because we do not fully understand the control of tumor DCs. To address this shortcoming, we performed scRNA-seq on human and mouse lung tumor lesions. Notably, we identified a distinct and nearly identical population of DC in both human and mouse lung tumors which upregulated genes associated with both DC maturation such as CD40 & IL12, and immunoregulation, including PD-L1 & CD200. This led us to annotate the cluster “mature DC enriched in immuno-regulatory molecules” (mregDC)(Maier et al. Nature 2020). Strikingly, mregDC were the DC carrying tumor antigen (Ag); meaning these DC are responsible for tumor Ag presentation. We hypothesize the immunostimulatory potential of tumor DC is dampened by genes upregulated in the mreg module and this thwarts induction of tumor immunity and response to αPD1. We propose that by targeting specific mreg genes we can decouple regulatory & stimulatory programs and enhance DC activation of tumor-reactive T cells and promote tumor immunity and αPD1 response. In support of our hypothesis, blocking signaling of IL4R, one of the upregulated genes in the mreg module, enhanced DC activation, expanded tumor-infiltrating T cells, and reduced tumor burden in a mouse model of NSCLC. Additionally, inhibition of Birc2/3, also upregulated in mregDC, led to substantially enhanced DC activation. To test our hypotheses and reach our objective, we will: (1) Determine the role of IL4R on induction of the mregDC state and tumor immunity. We will knockout IL4R in mouse & human DC and determine how this impacts mreg induction, Ag presentation, & tumor immunity. We will also combine anti-IL4R & anti-PD1 to assess synergy in controlling tumor growth in a preclinical model. (2) Evaluate Birc2/3 inhibition on the physiology, molecular state and immunostimulatory activity of intratumoral DC. We will test the hypothesis that pharmacological inhibition of Birc2/3 will enhance DC production of IL-12, as well co-stimulatory molecules, while facilitating cancer cell death and tumor Ag uptake, and result in robust tumor immunity. (3) Deconvolute the intrinsic regulators of DC phenotype and the mreg gene module. We will utilize a first-of-its-kind CRISPR genomics platform we developed to KO each of the 37 transcriptional related factor (TrF) genes upregulated by mregDC and determine how each impacts tumor DC activation and molecular state. The outcome of this project will provide a major advance in our understanding of intratumoral DC biology by determining the role of specific genes and pathways in dampening tumor DC functions, establish in preclinical models the therapeutic potential of compounds targeting two different pathways operating in mregDC, and identify additional molecules that could be targeted to enhance DC activity.

项目概要树突状细胞（DC）对肿瘤免疫具有重要影响。 DC 耗竭消除了肿瘤免疫并许多临床前肿瘤模型对 PD1 (αPD1) 免疫治疗的反应以及人类肿瘤中 DC 的浸润与临床结果呈正相关。不幸的是，我们仍然缺乏有效的策略来利用 DC 刺激肿瘤免疫，这在很大程度上是因为我们不完全了解肿瘤DC的控制。为了解决这个缺点，我们对人和小鼠肺部肿瘤病变进行了 scRNA 测序。尤其，我们在人和小鼠的肺部肿瘤中发现了一个不同且几乎相同的 DC 群体上调与 DC 成熟相关的基因，例如 CD40 和 IL12，以及免疫调节，包括 PD-L1 和 CD200。这导致我们将该簇注释为“富含免疫调节分子的成熟 DC” (mregDC)（Maier 等人，Nature 2020）。引人注目的是，mregDC 是携带肿瘤抗原（Ag）的 DC；意思是这些 DC 负责肿瘤 Ag 呈递。我们假设肿瘤 DC 的免疫刺激潜力是受到 mreg 模块中上调基因的抑制，这阻碍了肿瘤免疫和反应的诱导至αPD1。我们建议，通过针对特定的 mreg 基因，我们可以解耦监管和刺激程序增强DC对肿瘤反应性T细胞的激活，促进肿瘤免疫和αPD1反应。支持中根据我们的假设，阻断 IL4R（mreg 模块中上调基因之一）的信号传导，增强 DC 激活、扩大肿瘤浸润 T 细胞并减少 NSCLC 小鼠模型中的肿瘤负荷。此外，对 Birc2/3 的抑制（在 mregDC 中也上调）导致 DC 激活显着增强。为了检验我们的假设并达到我们的目标，我们将：（1）确定 IL4R 在诱导 mregDC 状态和肿瘤免疫。我们将敲除小鼠和人类 DC 中的 IL4R，并确定这是如何实现的影响 mreg 诱导、Ag 呈递和肿瘤免疫。我们还将结合抗IL4R和抗PD1来评估在临床前模型中控制肿瘤生长的协同作用。 (2)评估Birc2/3对生理的抑制作用，瘤内 DC 的分子状态和免疫刺激活性。我们将检验以下假设： Birc2/3 的药理学抑制将增强 DC 产生 IL-12 以及共刺激分子，同时促进癌细胞死亡和肿瘤Ag摄取，并产生强大的肿瘤免疫力。 (3) 解卷积 DC表型和mreg基因模块的内在调节因子。我们将利用首创的 CRISPR 我们开发的基因组学平台可敲除 37 个转录相关因子 (TrF) 基因上调的每一个基因 mregDC 并确定各自如何影响肿瘤 DC 激活和分子状态。该项目的成果通过确定特定的作用，将在我们对瘤内 DC 生物学的理解方面取得重大进展抑制肿瘤 DC 功能的基因和途径，在临床前模型中建立治疗潜力针对 mregDC 中运行的两种不同途径的化合物，并确定可以的其他分子有针对性地增强 DC 活性。