Interaction of cationic lipids with dendritic cells

阳离子脂质与树突状细胞的相互作用

• 批准号: 7581055
• 负责人: Leaf Huang
• 金额: $ 30.13万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-03-05 至 2013-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7581055
• 关键词: Adjuvant; Antigen Presentation; Antigen-Presenting Cells; Antigens; Biological Assay; Blood Vessels; Bone Marrow; CCL2 gene; CCL4 gene; CD80 gene; Cancer Vaccines; Cancer cell line; Cell membrane; Cells; Charge; Combined Modality Therapy; Combined Vaccines; Complementary DNA; Complex; Confocal Microscopy; DNA; DOTMA; Data; Dendritic Cells; Dendritic cell activation; Development; Distal; Dose; Drug Formulations; Encapsulated; Enzyme-Linked Immunosorbent Assay; Epitopes; Esters; Ethers; Event; Extracellular Signal Regulated Kinases; Fibroblast Growth Factor; Figs - dietary; Free Radical Scavengers; Glycerol; Goals; Granulocyte-Macrophage Colony-Stimulating Factor; Growth; Growth Factor Gene; Head; Human; Human Papillomavirus; Human papillomavirus 16; Immunity; Infiltration; Interleukin-12; Iodides; Knock-out; Label; Lead; Ligand Binding; Lipids; Liposomes; Lymphocyte; MAPK14 gene; MEKs; Malignant Neoplasms; Malignant neoplasm of cervix uteri; Mitogen-Activated Protein Kinases; Modeling; Modification; Mus; NADPH Oxidase; Neoplasm Metastasis; Oncogenes; Peptide Vaccines; Peptides; Phosphorylation; Play; Polyethylene Glycols; Production; Proteins; Reactive Oxygen Species; Reagent; Role; SB 203580; Series; Signal Pathway; Signal Transduction; Small Interfering RNA; Structure; Structure-Activity Relationship; Subcutaneous Injections; Surface; System; Testing; Therapeutic; Transfection; U-0126; Upper arm; Vaccines; Vascular Endothelial Cell; Vertebral column; Western Blotting; Work; abstracting; analog; base; chemokine; cytokine; design; follow-up; human MAPK14 protein; improved; inhibitor/antagonist; interest; mouse model; nanoparticle; neoplastic cell; novel; plasmid DNA; sigma receptors; therapeutic vaccine; tumor; upstream kinase

DESCRIPTION (provided by applicant): We have recently discovered that the cationic lipids DOTAP and DOEPC, originally developed as transfection reagents, can deliver a peptide antigen E7 to the dendritic cells (DC) and illicit a potent CTL activity against TC-1 tumor cells expressing the antigen. Signal transduction studies in the primary bone marrow-derived DC (BMDC) indicate that reactive oxygen species (ROS) is stimulated by DOTAP which leads to the phosphorylation and activation of two MAP kinases, ERK and p38,. pERK activation leads to the production and the secretion of potent chemokines, CCL2 and CCL4, by BMDC. P38 activation leads to the production of IL-12. Inhibition of pERK activation by specific inhibitors of Mek1/2, the upstream kinase which phosphorylates ERK, inhibited CCL2 production and, more importantly, the antitumor activity of DOTAP/E7. We hypothesize that ERK and p38 activation play a key role in the adjuvant activity of DOTAP, perhaps also DOEPC. The project will study in aim 1 the signaling of ROS and its downstream events leading to the production of CCL2 and IL-12. In aim 2, we will perform an extensive structure-activity relationship study of the cationic lipid to identify more active species that are suitable for vaccine formulation. Various DOTAP analogs will be synthesized and tested for ROS activation and vaccine activity. Finally in aim 3, we will attempt to improve the anti-tumor vaccine activity using the TC-1 tumor as a model for human cervical cancer. A number of formulation improvement strategies will be tested in the model. Furthermore, we will test the activity of siRNA against VEGF and other key oncogenes delivered by a targeted nanoparticle formulation to enhance the anti- tumor activity of the DOTAP/E7 vaccine. The goal of the study is to develop chemically well defined and potent cationic lipids for therapeutic vaccines against cancer. PUBLIC HEALTH RELEVANCE: The goal of the project is to develop an effective therapeutic vaccine for cervical cancer. The vaccine contains only two molecules, the antigen and a lipid, and is a very simple but highly active vaccine in a mouse model. Project will study the activity and the mechanism of the lipid adjuvant, in order to find more active lipid and more efficacious vaccine.

描述（由申请人提供）：我们最近发现阳离子脂质DOTAP和DOEPC，最初是作为转染试剂开发的，可以将肽抗原E7传递到树突状细胞（DC），并对表达抗原的TC-1肿瘤细胞具有有效的CTL活性。原发性骨髓源性DC （BMDC）的信号转导研究表明，DOTAP刺激活性氧（ROS），导致两种MAP激酶ERK和p38的磷酸化和激活。pERK激活导致BMDC产生和分泌强效趋化因子CCL2和CCL4。P38激活导致IL-12的产生。磷酸化ERK的上游激酶Mek1/2的特异性抑制剂抑制pERK激活，抑制CCL2的产生，更重要的是，抑制DOTAP/E7的抗肿瘤活性。我们假设ERK和p38的激活在DOTAP的佐剂活性中起关键作用，也可能是DOEPC。该项目将在目标1中研究ROS的信号传导及其下游事件导致CCL2和IL-12的产生。在目标2中，我们将对阳离子脂质进行广泛的构效关系研究，以确定更适合用于疫苗制剂的活性物种。各种DOTAP类似物将被合成并测试ROS激活和疫苗活性。最后，在目标3中，我们将尝试使用TC-1肿瘤作为人类宫颈癌的模型来提高抗肿瘤疫苗的活性。若干配方改进策略将在模型中进行测试。此外，我们将测试siRNA对VEGF和其他关键癌基因的活性，这些基因通过靶向纳米颗粒配方传递，以增强DOTAP/E7疫苗的抗肿瘤活性。该研究的目标是开发化学上定义良好且有效的阳离子脂质，用于治疗性癌症疫苗。公共卫生相关性：该项目的目标是研制一种有效的宫颈癌治疗疫苗。这种疫苗只含有两种分子，抗原和脂质，在小鼠模型中是一种非常简单但高度活跃的疫苗。本课题将对脂质佐剂的活性及其作用机制进行研究，以期找到更有活性的脂质和更有效的疫苗。