Immunosuppression and Metabolic Rewiring in Tumor-infiltrating Neutrophils

肿瘤浸润中性粒细胞的免疫抑制和代谢重组

• 批准号: 10637160
• 负责人: Xin Lu
• 金额: $ 36.2万
• 依托单位: UNIVERSITY OF NOTRE DAME
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-05-01 至 2028-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10637160
• 关键词: Ablation; Acceleration; Aconitic Acid; Address; Anti-Inflammatory Agents; Antibodies; Antioxidants; B-Lymphocytes; Biological Assay; Breast Cancer Model; Breast Cancer Treatment; Breast cancer metastasis; Carboxy-Lyases; Cell model; Cells; Clinical; Cytotoxic T-Lymphocytes; Development; Ensure; Enzymes; Face; Gene Silencing; Genes; Goals; Granulocyte-Macrophage Colony-Stimulating Factor; Human; Immune; Immune Evasion; Immunofluorescence Immunologic; Immunosuppression; Immunotherapeutic agent; Immunotherapy; In Vitro; Infiltration; Injections; JAK2 gene; Knockout Mice; Knowledge; Link; Lung; Macrophage; Malignant Neoplasms; Mammary Neoplasms; Mediating; Metabolic; Metastatic Neoplasm to the Lung; Metastatic breast cancer; Mitochondria; Modeling; Molecular; Morbidity - disease rate; Mouse Mammary Tumor Virus; Mus; Myeloid-derived suppressor cells; Neoplasm Metastasis; Neutrophil Infiltration; Outcome; Pathway interactions; Patients; Phenotype; Population; Production; Prognosis; Research; Resistance; Role; STAT3 gene; Safety; Sampling; Stains; T cell infiltration; T-Lymphocyte; Techniques; Testing; Therapeutic; Tissue Microarray; Transgenic Mice; Tumor Immunity; Up-Regulation; Validation; Wild Type Mouse; bench-to-bedside translation; bone; cancer immunotherapy; chemotherapy; curative treatments; deprivation; experimental study; follow-up; immune checkpoint blockade; immune resistance; immunoregulation; improved; inhibitor; loss of function; malignant breast neoplasm; mortality; mouse model; neutrophil; new therapeutic target; novel; pharmacologic; polyoma middle tumor antigen; pre-clinical; programmed cell death ligand 1; programs; response; single cell technology; single-cell RNA sequencing; synergism; therapeutic target; triple-negative invasive breast carcinoma; tumor; tumor microenvironment; validation studies

PROJECT SUMMARY/ABSTRACT Metastatic breast cancer (BC) remains largely resistant to immune checkpoint blockade (ICB) therapy. Tumor-infiltrating neutrophils (TINs) with immunosuppressive activity represent a major component in the tumor microenvironment (TME) to drive immunotherapy resistance. Therapeutic debilitation of immunosuppressive TINs is a promising approach to elicit synergistic efficacy when combined with immunotherapy. However, therapeutic targeting of TINs faces challenges in selectivity and safety, highlighted by the lack of TIN-specific targets. Recently, we used single-cell RNA sequencing to compare TINs and circulating neutrophils in BC models and identified aconitate decarboxylase 1 (Acod1) to be uniquely upregulated in TINs. Acod1 catalyzes the production of itaconate, a metabolite with anti-inflammatory activity in macrophages. But Acod1 function in neutrophils is poorly defined. Our preliminary results in BC mouse models suggest that TINs rely on Acod1 to sustain survival in the TME, and Acod1 loss leads to reduced TIN infiltration and metastasis. There are still significant knowledge gaps about the function, mechanism and therapeutic potential of Acod1 in TINs. Our central hypothesis is that Acod1 upregulated in TINs that infiltrate metastatic BC is essential for the TINs to persist in the TME and exert the immunosuppressive function, thus Acod1 ablation debilitates TINs, favors anti-tumor immunity, and sensitizes metastatic BC to immunotherapy. We propose to accomplish three Specific Aims: (Aim 1) Validate the pro-metastasis function of Acod1 in TINs in syngeneic and spontaneous murine mammary tumor models. (Aim 2) Identify the upstream and downstream molecular mechanisms underlying the upregulation and function of Acod1 in TINs. (Aim 3) Improve metastatic BC response to immunotherapy by Acod1 ablation and validate ACOD1 expression in clinical samples. To achieve our research goals, we have developed both whole-body and neutrophil-specific Acod1 knockout mice as hosts for BC syngeneic models, both mouse and human in vitro cell models of TINs, single- cell technologies for gene profiling and immune cell phenotyping, injection techniques that generate mammary tumors and metastases to lung and bone in the mice, and multiplex immunofluorescence staining techniques suitable for validation studies using clinical samples. We have assembled a strong research team with complementary expertise, which further ensures that the studies proposed are highly feasible to accomplish. Upon completion of the project, we expect to uncover the previous unknown function and mechanism of Acod1 in immunosuppressive TINs that are enriched in BC metastases, provide novel links between metabolic rewiring and immunoregulatory function of TINs, and generate the key preclinical evidence for targeting Acod1 to improve immunotherapy. In the long term, we envision that the bench-to-bedside translation of our findings through development of Acod1 inhibitors may accelerate the therapeutic application of combining agents that reprogram immunometabolism and immunotherapeutics to the curative treatment of BC and other cancers.

项目概要/摘要 转移性乳腺癌 (BC) 在很大程度上仍然对免疫检查点阻断 (ICB) 疗法具有抵抗力。 具有免疫抑制活性的肿瘤浸润性中性粒细胞 (TIN) 是肿瘤的主要成分 微环境（TME）驱动免疫治疗耐药性。免疫抑制剂治疗衰弱 TIN 是一种与免疫疗法结合时产生协同效应的有前途的方法。然而， TIN 的治疗靶向面临选择性和安全性方面的挑战，尤其是缺乏 TIN 特异性 目标。最近，我们使用单细胞 RNA 测序来比较 BC 模型中的 TIN 和循环中性粒细胞 并鉴定出乌头酸脱羧酶 1 (Acod1) 在 TIN 中独特地上调。 Acod1 催化 衣康酸的产生，衣康酸是巨噬细胞中具有抗炎活性的代谢物。但Acod1功能在 中性粒细胞的定义不明确。我们在 BC 小鼠模型中的初步结果表明，TIN 依赖 Acod1 维持 TME 中的生存，Acod1 缺失会导致 TIN 浸润和转移减少。还有 关于 Acod1 在 TIN 中的功能、机制和治疗潜力存在重大知识差距。 我们的中心假设是，Acod1 在浸润转移性 BC 的 TIN 中上调对于 TINs 持续存在于 TME 中并发挥免疫抑制功能，因此 Acod1 消融会削弱 TINs， 有利于抗肿瘤免疫，并使转移性 BC 对免疫治疗敏感。我们建议完成三项工作 具体目标：（目标 1）在同基因和自发性 TIN 中验证 Acod1 的促转移功能 小鼠乳腺肿瘤模型。 （目标2）识别上游和下游分子机制 TIN 中 Acod1 的上调和功能的基础。 （目标 3）改善转移性 BC 反应 通过 Acod1 消融进行免疫治疗并验证临床样本中的 ACOD1 表达。 为了实现我们的研究目标，我们开发了全身和中性粒细胞特异性 Acod1 敲除小鼠作为 BC 同系模型的宿主，包括 TIN 的小鼠和人类体外细胞模型、单 用于基因分析和免疫细胞表型分析的细胞技术、产生乳腺的注射技术 小鼠肿瘤和肺和骨转移，以及多重免疫荧光染色技术 适用于使用临床样本的验证研究。我们组建了一支强大的研究团队 互补的专业知识，进一步确保拟议的研究具有高度的可行性。 项目完成后，我们期望揭开之前未知的功能和机制 免疫抑制 TIN 中的 Acod1 在 BC 转移中富集，提供了代谢之间的新联系 TIN 的重新布线和免疫调节功能，并生成靶向 Acod1 的关键临床前证据 以改善免疫治疗。从长远来看，我们预计我们的研究结果可以从实验室转化为临床 通过Acod1抑制剂的开发可能会加速联合药物的治疗应用 重新规划免疫代谢和免疫治疗，以治愈 BC 和其他癌症。