Deciphering the molecular control of intratumoral dendritic cells

破译瘤内树突状细胞的分子控制

• 批准号: 10331052
• 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
• 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/10331052
• 关键词: ALDH1A2 gene; Address; Antigen Presentation; Antigens; Antitumor Response; Apoptosis; 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; Consequentialism; Data; Dendritic Cells; Dendritic cell activation; Dendritic cell tumor; Equilibrium; Genes; Genetic Transcription; Goals; Human; IL12B gene; IL4 gene; IL4R gene; Immune; Immunity; Immunophenotyping; Immunotherapy; Individual; Infiltration; Interleukin-12; Interleukin-4; Knock-out; Lead; Lesion; Lung Neoplasms; Lymphocyte Activation; Mediating; Modeling; Molecular; Molecular Profiling; Mus; NF-kappa B; Nature; Non-Small-Cell Lung Carcinoma; Outcome; Output; PD-1 blockade; Pathway Analysis; Pathway interactions; Pharmacology; 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 Immunity; anti-PD-1; base; cancer cell; cancer immunotherapy; cytokine; draining lymph node; effector T cell; genomic platform; immunoregulation; improved; in vivo; inhibitor; interest; macrophage; mouse model; 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

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.

项目摘要 树突状细胞（DC）对肿瘤免疫史有重大影响。 DC消耗废除了肿瘤免疫史和 在许多临床前肿瘤模型中对PD1（αPD1）免疫疗法的反应和人类肿瘤的直流浸润 是临床结果的正相关。不幸的是，我们仍然缺乏利用DC的有效策略 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。 为了解决这一缺点，我们对人和小鼠肺肿瘤病变进行了SCRNA-SEQ。尤其， 我们在人和小鼠肺肿瘤中确定了DC的独特且几乎相同的人群 与DC成熟相关的上调基因，例如CD40和IL12，以及免疫调节，包括 PD-L1和CD200。这导致我们注释了“成熟的直流免疫调节分子”的簇。 （MREGDC）（Maier等人自然2020）。令人惊讶的是，MREGDC是携带肿瘤抗原（AG）的直流；意思是这些 DC负责肿瘤AG表现。我们假设肿瘤直流的免疫刺激潜力 被MREG模块中更新的基因抑制，这种阻碍了肿瘤免疫和反应的诱导 到αPD1。我们建议，通过靶向特定的MREG基因，我们可以将监管和刺激程序解除 并增强肿瘤反应性T细胞的直流激活，并促进肿瘤免疫组织化学和αPD1反应。支持 在我们的假设中，IL4R的信号是MREG模块中更新的基因之一，增强的DC 在NSCLC的小鼠模型中，激活，膨胀肿瘤浸润的T细胞以及减少肿瘤燃烧。 另外，在MREGDC中也更新的BIRC2/3的抑制作用导致DC激活大大增强。 为了检验我们的假设并实现我们的目标，我们将：（1）确定IL4R在诱导的作用 MREGDC状态和肿瘤免疫。我们将在鼠标和人类DC中淘汰IL4R，并确定如何 影响MREG诱导，AG表现和肿瘤免疫学。我们还将结合抗IL4R和抗PD1来评估 临床前模型中控制肿瘤生长的协同作用。 （2）评估BIRC2/3对生理学的抑制作用， 肿瘤内DC的分子状态和免疫刺激活性。我们将检验以下假设 BIRC2/3的药理抑制作用将增强IL-12的直流产生，而共刺激性分子，而不是共同刺激的分子 促进癌细胞死亡和肿瘤AG摄取，并导致稳健的肿瘤免疫力。 （3）反应 DC表型和MREG基因模块的内在调节剂。我们将利用第一范围的CRISPR 我们开发了基因组平台为37个转录相关因素（TRF）基因中的每个KO开发 MregDC并确定每个如何影响肿瘤直流激活和分子状态。这个项目的结果 通过确定特定的作用 型号肿瘤直流功能中的基因和途径，在临床前模型中建立治疗潜力 针对MREGDC运行的两种不同途径的化合物，并识别可能的其他分子 针对增强直流活性。