Mechanisms of B cell specific IL-35 expression in cancer.

癌症中 B 细胞特异性 IL-35 表达的机制。

• 批准号: 9910621
• 负责人: Daniel E Michaud
• 金额: $ 3.39万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-01-01 至 2021-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9910621
• 关键词: Adaptor Signaling Protein; Address; Adoptive Transfer; Agammaglobulinaemia tyrosine kinase; Antigens; Autoimmune Diseases; B-Cell Activation; B-Lymphocytes; CD19 gene; CD8-Positive T-Lymphocytes; Cancer Etiology; Cell Line; Cell Nucleus; Cells; Cellular biology; Cessation of life; Data; Diagnosis; Disease; Endosomes; Exposure to; Goals; Growth; Immune; Immunosuppression; Immunotherapeutic agent; Immunotherapy; Implant; In Vitro; Infiltration; Injections; Interleukins; Malignant Neoplasms; Malignant neoplasm of pancreas; Mediating; Modality; Modeling; Mus; MyD88 protein; Pancreatic Ductal Adenocarcinoma; Pathogenicity; Pathway interactions; Patient-Focused Outcomes; Patients; Phosphotransferases; Production; Receptor Activation; Receptor Cell; Receptor Protein-Tyrosine Kinases; Receptor Signaling; Receptors, Antigen, B-Cell; Regulation; Research; Research Proposals; Signal Pathway; Signal Transduction; Signaling Molecule; Specificity; Structure of beta Cell of islet; Survival Rate; T cell response; T-Lymphocyte; TLR2 gene; TLR7 gene; Testing; Therapeutic; Toll-Like Receptor Pathway; Toll-like receptors; Training; Tumor Immunity; United States; Up-Regulation; Western Blotting; anti-tumor immune response; base; cancer therapy; career; cytokine; design; experience; immunoregulation; improved; in vivo; in vivo monitoring; inhibitor/antagonist; insight; mouse model; neutralizing antibody; novel; pancreatic neoplasm; pancreatic tumorigenesis; treatment response; tumor; tumor growth; tumor microenvironment; tumorigenic

Project Abstract Pancreatic Ductal Adenocarcinoma (PDA) is the most commonly diagnosed form (~90%) of pancreatic cancer with a dismal 5-year survival rate of ~8%. PDA features a prominent pro-tumorigenic microenvironment rich in immunosuppressive cells that inhibit functions of anti-tumor immunity. Current immunotherapeutic approaches have been unsuccessful in improving PDA patient outcomes, leaving a need for a better understanding of the immunomodulatory signaling mechanisms within pancreatic tumor microenvironment. We have identified infiltrating regulatory B cells as being key contributors to pancreatic tumorigenesis through their expression of the regulatory cytokine IL-35, which directly inhibits anti-tumor immune responses. The mechanisms that cause B cells to produce IL-35 in cancer and promote pancreatic tumor growth are not known. Using a mouse model harboring a monoclonal fixed B cell receptor (BCR), or mice with a B cell-specific deletion of the Toll-Like Receptor (TLR) adaptor protein MyD88, I found that orthotopically implanted pancreatic tumors were drastically reduced in size as compared to WT mice. I have also conducted in vitro analysis of TLR and BCR activation and found that co-stimulation of endosomal TLRs and the BCR leads to a robust increase of IL-35 expression in B cells. Based on my preliminary data, I hypothesize that cancer-driven upregulation of IL-35 in B cells is dependent on crosstalk between BCR and endosomal TLRs, which promotes pancreatic tumor growth through immunosuppression. I propose two specific aims to test my hypothesis. In Aim 1, I will determine how BCR and endosomal TLR signaling via Bruton's tyrosine kinase (BTK) contributes to expression of IL-35 expression in B cells, using analysis of signaling pathways in primary regulatory B cells and established B cell lines. In Aim 2, I will investigate how activation of both BCR and TLR signaling in B cells promotes PDA tumor growth in vivo. To accomplish this task, I will analyze mouse models expressing a fixed BCR specificity with or without specific antigen exposure, mouse models lacking MyD88 signaling in B cells, as well as a cross of the two models. Our proposed research will provide an understanding of a previously uncharacterized facet of B cell-mediated function in PDA and use state-of-the-art PDA murine models to test strategies that block immune suppressive pathways. Ultimately, I anticipate my findings will reveal a targetable mechanism to inhibit B cell-mediated immunosuppression in pancreatic tumors and provide a multi-faceted training experience to help advance my scientific career.

项目摘要 胰腺导管腺癌（PDA）是最常见的胰腺癌（约90%） 5年生存率只有8% PDA具有显著的促肿瘤发生微环境， 抑制抗肿瘤免疫功能的免疫抑制细胞。目前的免疫方法 在改善PDA患者结局方面一直不成功，因此需要更好地了解 胰腺肿瘤微环境中的免疫调节信号机制。我们已经确定 浸润性调节性B细胞通过表达 调节细胞因子IL-35，其直接抑制抗肿瘤免疫应答。导致 目前尚不清楚B细胞在癌症中产生IL-35并促进胰腺肿瘤生长。使用小鼠模型 携带单克隆固定的B细胞受体（BCR）的小鼠，或具有Toll样受体的B细胞特异性缺失的小鼠， 受体（TLR）衔接蛋白MyD 88，我发现原位种植胰腺肿瘤， 与WT小鼠相比，其尺寸减小。我还进行了TLR和BCR激活的体外分析， 发现内体TLR和BCR的共刺激导致B中IL-35表达的强烈增加 细胞基于我的初步数据，我假设癌症驱动的B细胞中IL-35的上调依赖于 BCR和内体TLR之间的串扰，通过 免疫抑制我提出两个具体目标来检验我的假设。在目标1中，我将确定BCR和 通过布鲁顿酪氨酸激酶（BTK）的内体TLR信号传导有助于B中IL-35表达的表达 细胞，使用原代调节B细胞和建立的B细胞系中的信号传导途径的分析。在目标2中，我 将研究B细胞中BCR和TLR信号的激活如何促进体内PDA肿瘤的生长。到 为了完成这项任务，我将分析表达固定BCR特异性的小鼠模型， 抗原暴露、在B细胞中缺乏MyD 88信号传导的小鼠模型以及两种模型的交叉。我们 拟议的研究将提供一个以前未表征的方面的理解B细胞介导的 功能，并使用最先进的PDA小鼠模型来测试阻断免疫抑制的策略。 途径。最终，我期望我的研究结果将揭示一种靶向机制，以抑制B细胞介导的 胰腺肿瘤中的免疫抑制，并提供多方面的培训经验，以帮助提高我的 科学生涯。