Dendritic Cell Proteoglycans and Reprogramming Cancer Immunity

树突状细胞蛋白聚糖和重编程癌症免疫

• 批准号: 10045943
• 负责人: MARK M FUSTER
• 金额: --
• 依托单位: VA SAN DIEGO HEALTHCARE SYSTEM
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-10-01 至 2022-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10045943
• 关键词: Activities of Daily Living; Address; Affect; Anabolism; Antigen Presentation; Antigens; Antisense Oligonucleotides; Area; Binding; CCL21 gene; CD8-Positive T-Lymphocytes; CX3CL1 gene; CXCL12 gene; Cancer Model; Cancer Patient; Carbohydrates; Carcinoma; Cell Communication; Cell Maturation; Cell physiology; Cell surface; Cells; Chest; Clinical; Complex; Cytolysis; Dendrites; Dendritic Cells; Dendritic cell activation; Dendritic cell tumor; Development; Disease remission; Drug Design; Enzymes; Equilibrium; Eye; Genetic; Goals; Growth; Heparan Sulfate Biosynthesis; Heparitin Sulfate; Human; Immune; Immune Tolerance; Immunity; Immunologics; Immunophenotyping; Immunosuppression; Immunotherapy; KRAS2 gene; Kinetics; Lead; Lewis Lung Carcinoma; Lewis lung carcinoma cell; Lung Neoplasms; Lymphatic; Lymphatic Endothelium; Lymphocyte; Malignant Neoplasms; Malignant neoplasm of lung; Mediating; Methods; Modeling; Molecular; Mus; Mutation; Neoplasm Metastasis; Ovalbumin; Ovum; Pathologic; Pathway interactions; Patients; Phenotype; Polysaccharides; Preparation; Primary Neoplasm; Production; Property; Proteoglycan; Regulatory T-Lymphocyte; Signal Transduction; Sulfate; Surface; T cell response; T-Lymphocyte; Testing; Therapeutic; Transgenic Organisms; Translations; Tumor Antigens; Tumor Immunity; Ursidae Family; Validation; Work; anti-cancer; base; cell behavior; cell motility; chemokine; chemokine receptor; cytokine; draining lymph node; effector T cell; immune function; improved; inhibitor/antagonist; lung Carcinoma; lymphatic vessel; lymphocyte proliferation; migration; mimetics; mutant; neoplastic cell; novel; novel strategies; novel therapeutics; pre-clinical; programs; proteoglycan core protein; receptor; response; scaffold; small molecule inhibitor; sugar; syndecan; targeted agent; trafficking; tumor; tumor growth; tumor microenvironment; tumor progression

Early in cancer growth, metastases and dendritic cells (DCs) traffic to draining lymph nodes (DLN). In lung cancer and other carcinomas, immune-suppression dominates a tumor microenvironment characterized by immature DCs, weak T-effector responses, proliferation of T-regulatory cells (Tregs), and over-production of immuno-suppressive cytokines. This heavily represses anti-tumor immunity. A high-impact area of new scientific discovery is immunotherapy. We find dramatic clinical responses in some advanced-stage cancer patients by blocking suppression of effector T cells. Only a fraction (<25%) of such patients, however, respond with durable remissions. Preliminary work shows that a unique class of glycans known as heparan sulfate (HS) drives subversive DC traffic and DC immaturity. While we have inhibited such DC traffic by targeting lymphatic endothelial HS, new work suggests that DC-specific HS alterations may modulate both pathologic chemokine- dependent DC traffic as well as DC maturation and function. New studies also show that these properties may favorably impact anti-tumor T cell functions, with inhibition of tumor growth and progression. Herein we target such glycans in tumor and cell-based studies, while studying immune-function and molecular mechanisms. This proposal addresses the hypothesis that targeting HS glycans on the surface of DCs in lung cancer through genetic means and novel inhibitors will inhibit DLN colonization by tolerogenic DCs and improve anti- tumor immunity. Reduced immune-tolerance and improved tumor-antigen responses by more mature DCs, with improved T cell induction will result in a novel endogenous anti-tumor state. To test this, we propose to: (1) Characterize tumor growth and anti-tumor immunity in model antigen- as well as spontaneous lung carcinoma models in mice bearing DC-glycan alterations. We will assess how a DC-targeted mutation in a key sulfating HS biosynthetic enzyme (Ndst1) affects T cell immunity in tumors and thoracic DLN of mice with orthotopic Ovalbumin-expressing Lewis lung carcinomas (LLC-Ova), including Ova-specific immunity and effects on tumor growth. Immunity and tumor growth in a KRAS transgenic mutant model will also be studied. (2) Study anti-tumor DC and T cell functions in ex-vivo preparations from lung carcinoma bearing mice with DC-specific alterations in HS biosynthesis. DC maturation, antigen presentation, and the capacity of DCs from LLC-Ova tumors grown in DC-targeted HS mutants to activate Ova-sensitized T cells will be examined, as will tumor-cytolytic capacity of CD8+ T cells isolated from DLNs of tumor-bearing mutant vs control mice. Studies will also include the effects of mutation on DLN colonization by plasmacytoid DCs and their functional capacity. (3) Assess chemokine-receptor interactions and signaling mechanisms in the setting of DC-targeted alterations in HS biosynthesis; and the effects of novel HS inhibitors on tumor growth and immunity. We will study how HS mutation affects DC-surface binding of lymphatic chemokines CCL21, CXCL12, or CX3CL1 to cognate receptors. The effect of HS mutation on DC migration and maturation signaling in response to chemokine will be assessed, including the activation of pDCs through novel TLR-dependent mechanisms. Finally, we will study how novel HS inhibitors affect signaling by tumor DCs as well as lung carcinoma growth and immunity. This work examines novel mechanisms whereby altering trafficking and maturation of tumor DCs may effectively re-program endogenous anti-tumor immunity. The genetic target validation and rational introduction of glycan-targeting inhibitors may guide new therapies to achieve immune eradication of human lung cancer.

在癌症生长早期，转移和树突状细胞（DC）运输至引流淋巴结（DLN）。肺 癌症和其他癌症，免疫抑制主导肿瘤微环境，其特征在于 不成熟的DC、弱的T效应子应答、T调节细胞（Tcells）的增殖和 免疫抑制细胞因子。这严重抑制了抗肿瘤免疫。一个新的高影响力领域 科学发现是免疫疗法我们在一些晚期癌症中发现了戏剧性的临床反应 患者通过阻断效应T细胞的抑制。然而，只有一小部分患者（<25%）对 持久缓解。初步工作表明，一类独特的聚糖称为硫酸乙酰肝素（HS） 驱动颠覆性的DC流量和DC不成熟。虽然我们已经通过靶向淋巴细胞来抑制这种DC运输， 内皮HS，新的工作表明，DC特异性HS改变可能调节病理性趋化因子- 依赖的DC业务以及DC成熟和功能。新的研究还表明，这些特性可能 有利地影响抗肿瘤T细胞功能，抑制肿瘤生长和进展。在此，我们的目标 在肿瘤和细胞研究中使用这种聚糖，同时研究免疫功能和分子机制。 这一提议提出了一个假设，即靶向肺癌中DC表面的HS聚糖 通过遗传手段和新的抑制剂将抑制DLN通过致耐受性DC的定殖，并改善抗- 肿瘤免疫通过更成熟的DC降低免疫耐受性和改善肿瘤抗原应答， 具有改善的T细胞诱导的药物组合物将导致新的内源性抗肿瘤状态。为了检验这一点，我们建议： (1)表征模型抗原和自发肺中的肿瘤生长和抗肿瘤免疫 在携带DC-聚糖改变的小鼠的癌模型中。我们将评估一个关键基因中的DC靶向突变 硫酸化HS生物合成酶（Ndst 1）影响肿瘤和胸DLN小鼠的T细胞免疫 原位卵清蛋白表达刘易斯肺癌（LLC-Ova），包括Ova特异性免疫和 对肿瘤生长的影响还将研究KRAS转基因突变体模型中的免疫和肿瘤生长。 (2)肺癌荷瘤小鼠DC和T细胞体外抗肿瘤作用的研究 HS生物合成中的DC特异性改变。DC成熟、抗原提呈和DC的能力， 将检查在DC靶向HS突变体中生长以激活Ova致敏T细胞的LLC-Ova肿瘤， 从携带肿瘤的突变体与对照小鼠的DLN分离的CD 8 + T细胞的肿瘤细胞溶解能力。研究 还将包括突变对浆细胞样DC的DLN定殖及其功能能力的影响。 (3)评估DC靶向改变背景下的趋化因子-受体相互作用和信号传导机制 在HS生物合成中的作用;以及新型HS抑制剂对肿瘤生长和免疫的影响。我们将研究如何HS 突变影响淋巴趋化因子CCL 21、CXCL 12或CX 3CL 1与同源物的DC表面结合 受体。HS突变对DC迁移和成熟信号转导的影响将响应趋化因子， 包括通过新的TLR依赖性机制激活pDC。最后我们将 研究新型HS抑制剂如何影响肿瘤DC的信号传导以及肺癌生长和免疫。 这项工作探讨了改变肿瘤DC的运输和成熟的新机制， 有效地重新编程内源性抗肿瘤免疫。遗传目标的验证与合理引入 聚糖靶向抑制剂的研究可能会指导新的治疗方法，以实现免疫根除人类肺癌。