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.

项目摘要/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)治疗耐药。 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)导致免疫治疗耐药。免疫抑制的治疗性衰弱 TINS是一种很有前途的方法，当与免疫治疗相结合时，可以产生协同疗效。然而， 锡靶向治疗面临选择性和安全性方面的挑战，突出表现为缺乏锡特异性 目标。最近，我们使用单细胞rna测序来比较bc模型中的TINS和循环中性粒细胞。 并鉴定出在TINS中唯一上调的乌头酸脱羧酶1(Acod1)。Acod1催化 巨噬细胞中具有抗炎活性的代谢物衣康酸的产生。但Acod1在 中性粒细胞的定义并不明确。我们在BC小鼠模型中的初步结果表明TINS依赖于Acod1来 维持在TME中的生存，Acod1的缺失导致锡的渗透和转移减少。仍然有 关于TINS中Acod1的功能、机制和治疗潜力的显著知识空白。 我们的中心假设是，在TIN中上调的Acod1对转移的BC是必不可少的。 TINS在TME中持续存在并发挥免疫抑制作用，因此Acod1消融使TINS衰弱， 有利于抗肿瘤免疫，并使转移性BC对免疫治疗敏感。我们计划完成三个目标 具体目的：(目的1)验证Acod1在同基因和自发性TINS中的促转移功能 小鼠乳腺肿瘤模型。(目标2)确定上下游分子机制 TINS中Acod1的上调和功能。(目标3)改善转移性BC对 通过Acod1消融进行免疫治疗，并验证Acod1在临床样本中的表达。 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的Acod1 以基因敲除小鼠为宿主的BC同基因模型，包括小鼠和人的TINS体外细胞模型，单核细胞 基因图谱和免疫细胞表型的细胞技术，产生乳房的注射技术 小鼠肺、骨肿瘤转移及多重免疫荧光染色技术 适用于使用临床样本进行验证研究。我们已经组建了一支强大的研究团队， 专家意见相辅相成，进一步确保拟议的研究具有很高的可行性。 项目完成后，我们预计将揭开之前未知的功能和机制 富含BC转移的免疫抑制TIN中的Acod1在代谢之间提供了新的联系 TINS的重组和免疫调节功能，并为靶向Acod1提供关键的临床前证据 提高免疫治疗水平。从长远来看，我们设想将我们的研究结果从长椅到床边的翻译 通过开发Acod1抑制剂可以加速联合用药的治疗应用 重新编程免疫代谢和免疫疗法，以治疗BC和其他癌症。