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

基本信息

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
项目类别：
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

项目摘要

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）：原发性恶性脑肿瘤的预后令人沮丧，其中四分之一的患者在诊断五年以上存活（1）。尤其是胶质母细胞瘤细胞具有剧烈的侵入性，通常从肿瘤切除腔迁移到雄辩的大脑中（2）。由于治疗不足，高级神经胶质瘤需要替代细胞特异性治疗，例如免疫疗法。迄今为止，几项针对神经胶质瘤的免疫疗法试验已显示出令人鼓舞的结果（3-6），但我们缺乏有关如何可重复安装抗肿瘤免疫反应的知识。超过1,000次癌症疫苗试验已经并且继续涉及肿瘤细胞作为抗原来源的培养。直到我们最近发现5％O2中培养的发现增加的发现，氧气对培养肿瘤细胞的免疫原性的影响知之甚少：（1）与神经胶质瘤相关的抗原表达（参考文献8）；（2）神经胶质瘤裂解物的辅助性（7，8）。这些发现导致了蛋白质的鉴定膜联蛋白A2（此后“ A2”），一种富含5％O2的神经胶质瘤细胞中的新型免疫佐剂。我假设A2是与损伤相关的分子模式，并确定它是有效的Toll样受体2（TLR2）配体。 A2的TLR2活性除了从5％O2的裂解物疫苗的小鼠中强大的CD8 T细胞反应外，还导致了A2增加肿瘤抗原的交叉表现。体外数据表明，A2在树突状细胞（DC）上增强了交叉表现，这些细胞能够启动抗肿瘤CD8 T细胞。但是，确切的机制尚不清楚。为了检验该假设，该提案将分为两个独立的特定目的：（1）研究膜联蛋白A2对抗原摄取，路由和蛋白水解的影响：a。测量DC中标记的抗原和肿瘤裂解物的吸收； b。使用DC的显微镜确定抗原的亚细胞位置；和c。量化DC中的蛋白酶体活性和组成。（2）确定A2是否增强了T细胞受体信号传导，肿瘤反应性CD8 T细胞的膨胀以及延伸含有神经胶质瘤小鼠的存活率：a。在神经胶质瘤反应性CD8 T细胞中量化相对T细胞受体信号强度； b。测量内源性神经胶质瘤反应性CD8 T细胞的扩展c。确定A2/神经胶质瘤肽疫苗是否扩展了含有神经胶质瘤的小鼠的存活，并且生存率是否与离体CD8 T细胞反应相关。这些研究的完成，基本科学癌和免疫学培训以及专注于癌症治疗的医学院培训将使我为脑肿瘤免疫疗法的研究计划做好准备。