Annexin A2 as a Novel Enhancer of Tumor-Associated Antigen Processing

膜联蛋白 A2 作为肿瘤相关抗原加工的新型增强剂

• 批准号: 8659972
• 负责人: Brian Magne Andersen
• 金额: $ 4.77万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-05-03 至 2016-05-02
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8659972
• 关键词: ANXA2 gene; Adjuvant; Adjuvanticity; Affect; Annexins; Antigens; Basic Science; Binding; Brain; Brain Neoplasms; CD8B1 gene; Cancer Immunology Science; Cancer Vaccines; Cell Culture Techniques; Cells; Clinical; Clinical Trials; Cross Presentation; Cross-Priming; Cultured Tumor Cells; Data; Dendritic Cells; Diagnosis; Disease; Doctor of Philosophy; Endosomes; Enhancers; Environment; Glioblastoma; Glioma; Goals; Human; Immune response; Immunologic Adjuvants; Immunotherapy; In Vitro; Injection of therapeutic agent; Knowledge; Label; Left; Ligands; Location; Malignant Glioma; Malignant Neoplasms; Malignant neoplasm of brain; Measures; Memory; Methods; Microscopy; Modeling; Molecular; Mus; Oncologist; Operative Surgical Procedures; Outcome; Oxygen; Pathway interactions; Patients; Pattern; Peptides; Phospholipids; Physicians; Poly ICLC; Process; Property; Proteins; Proteolysis; Radiosurgery; Receptor Signaling; Relative (related person); Research; Scientist; Signal Transduction; Source; Surgically-Created Resection Cavity; T cell response; T-Cell Receptor; T-Lymphocyte; TLR3 gene; Techniques; Testing; Tissues; Toll-Like Receptor 2; Training; Tumor Antigens; Tumor Expansion; United States; Ursidae Family; Vaccination; Vaccine Adjuvant; Vaccine Therapy; Vaccines; Vesicle; antigen processing; base; cancer therapy; career; immunogenicity; improved; medical schools; multicatalytic endopeptidase complex; novel; outcome forecast; peptide I; programs; public health relevance; research study; response; standard of care; temozolomide; tumor; uptake

DESCRIPTION (provided by applicant): Primary malignant brain tumors bear dismal prognoses, with one quarter of patients surviving beyond five years of diagnosis (1). Glioblastoma cells in particular are fiercely invasive, often migrating several centimeters from the tumor resection cavity into eloquent brain (2). Due to inadequate treatment, high-grade gliomas require alternate, cell-specific treatments such as immunotherapy. Thus far several immunotherapy trials for glioma have shown promising results (3-6), but we lack knowledge on how to reproducibly mount an anti-tumor immune response. Over 1,000 cancer vaccine trials have and continue to involve the culture of tumor cells as the antigen source. Little was known on the influence of oxygen on the immunogenicity of cultured tumor cells until our recent findings that culture in 5% O2 increases: (1) glioma-associated antigen expression (Ref 8); and (2) adjuvanticity of glioma lysates (7, 8). These findings led to the identification of the protein annexin A2 ("A2" hereafter), a novel immune adjuvant enriched in glioma cells grown in 5% O2. I hypothesized that A2 is a damage-associated molecular pattern and determined that it is a potent toll-like receptor 2 (TLR2) ligand. A2's TLR2 activity, in addition to the powerful CD8 T cell responses in mice given lysate vaccines from 5% O2, led to the hypothesis that A2 increases cross presentation of tumor antigens. In vitro data indicate that A2 boosts cross presentation on dendritic cells (DCs), which are capable of priming anti-tumor CD8 T cells; however, the precise mechanisms are unclear. To test this hypothesis, this proposal will be divided into two separate specific aims: (1) Investigate annexin A2's effect on antigen uptake, routing, and proteolysis: a. Measure uptake of labeled antigens and tumor lysate in DCs; b. Determine the subcellular location of antigen using microscopy of DCs; and c. Quantify proteasome activity and composition in DCs. (2) Determine if A2 potentiates T cell receptor signaling, expansion of tumor-reactive CD8 T cells, and extends survival of glioma-bearing mice: a. Quantify relative T cell receptor signal strength in glioma-reactive CD8 T cells;b. Measure expansion of endogenous glioma-reactive CD8 T cells c. Determine if A2/glioma peptide vaccination extends survival of glioma-bearing mice, and if survival correlates with ex vivo CD8 T cell response. Completion of these studies, basic-science cancer and immunology training, and medical school training focused on cancer therapy will prepare me for a career as a physician-scientist with a research program in brain tumor immunotherapy.

描述（由申请人提供）：原发性恶性脑肿瘤具有令人沮丧的预后，四分之一的患者在诊断后存活超过五年（1）。特别是胶质母细胞瘤细胞具有强烈的侵袭性，通常从肿瘤切除腔迁移几厘米进入功能脑（2）。由于治疗不足，高级别胶质瘤需要替代的细胞特异性治疗，如免疫治疗。到目前为止，针对胶质瘤的几项免疫治疗试验已经显示出有希望的结果（3-6），但是我们缺乏关于如何可重复地建立抗肿瘤免疫应答的知识。超过1，000项癌症疫苗试验已经并将继续涉及肿瘤细胞作为抗原来源的培养。我们对氧气对培养肿瘤细胞免疫原性的影响知之甚少，直到我们最近发现在5%O2中培养增加了：（1）胶质瘤相关抗原表达（参考文献8）;（2）胶质瘤裂解物的佐剂性（7，8）。这些发现导致了蛋白质的鉴定 膜联蛋白A2（下文称为“A2”），一种在5%O2中生长的神经胶质瘤细胞中富集的新型免疫佐剂。我假设A2是一种损伤相关的分子模式，并确定它是一种有效的Toll样受体2（TLR 2）配体。A2的TLR 2活性，除了在给予来自5%O2的裂解物疫苗的小鼠中的强大的CD 8 T细胞应答之外，导致了A2增加肿瘤抗原的交叉呈递的假设。体外数据表明，A2促进树突状细胞（DC）上的交叉呈递，其能够引发抗肿瘤CD 8 T细胞;然而，确切的机制尚不清楚。为了检验这一假设，该提议将分为两个单独的具体目标：（1）研究膜联蛋白A2对抗原摄取、路由和蛋白水解的作用：测量DC中标记抗原和肿瘤裂解物的摄取; B.使用DC的显微镜检查确定抗原的亚细胞位置;以及c.定量DC中的蛋白酶体活性和组成。 (2)确定A2是否增强T细胞受体信号传导、肿瘤反应性CD 8 T细胞的扩增以及延长具有神经胶质瘤的小鼠的存活：定量神经胶质瘤反应性CD 8 T细胞中的相对T细胞受体信号强度;B.测量内源性神经胶质瘤反应性CD 8 T细胞的扩增。确定A2/神经胶质瘤肽疫苗接种是否延长了荷神经胶质瘤小鼠的存活，以及存活是否与离体CD 8 T细胞应答相关。完成这些研究，基础科学癌症和免疫学培训，以及专注于癌症治疗的医学院培训将为我作为一名医生科学家的职业生涯做好准备，并在脑肿瘤免疫治疗方面开展研究计划。