Cancer Therapeutics that Anchor Proteins to Membranes

将蛋白质锚定在膜上的癌症治疗方法

基本信息

批准号：
7172627
负责人：
BLAKE PETERSON
金额：
$ 23.19万
依托单位：
PENNSYLVANIA STATE UNIVERSITY, THE
依托单位国家：
美国
项目类别：
财政年份：
2000
资助国家：
美国
起止时间：
2000-01-01 至 2008-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7172627
关键词：
Ablation Acids Address Allogeneic Bone Marrow Transplantation Antigen-Presenting Cells Antigens Avidin Binding Biotin CD8B1 gene CD95 Antigens Cancer Model Cancer cell line Catalytic Domain Cell Line Cell membrane Cell physiology Cells Cessation of life Chemotherapy-Oncologic Procedure Chimera organism Chimeric Proteins Cholera Toxin Cholesterol Clathrin Collaborations Complex Cultured Cells Cytosol Cytotoxin Daunorubicin Effectiveness Endocytosis Endosomes Evaluation Exotoxins Ganglioside GM1 Genes Grant Green Fluorescent Proteins Hematopoietic Histocompatibility Antigens Class I Human Hydrazones Immune response Immunization Immunologic Adjuvants Immunotherapy In Vitro Ligands Link Lymphocyte MHC antigen Malignant Neoplasms Mammalian Cell Mediating Membrane Membrane Microdomains Molecular Multi-Drug Resistance Mus Neuroblastoma Neuropeptide Y Receptor Numbers Ovalbumin P-Glycoprotein Penetration Peptide/MHC Complex Pharmaceutical Preparations Play Process Production Progress Reports Proteins Proteolytic Processing Rate Recruitment Activity Regulation Role Signal Transduction Sphingolipids Stem cells Streptavidin Surface System T-Cell Activation T-Lymphocyte Testing Therapeutic Toxin Transgenic Mice Tumor Antigens Universities analog antitumor agent base cancer cell cancer therapy caveolin 1 cell growth cell mediated immune response chemotherapy cholesterol-binding protein cholesterylamine cytotoxic graft vs host disease in vivo macrophage medical schools mouse model neoplastic cell neuropeptide Y novel novel strategies pre-clinical receptor small molecule tool tumor tumor xenograft vaccine development

项目摘要

Multidrug resistant (MDR) cancer remains the primary impediment to curative cancer chemotherapy. MDR cancer cells differ from typical tumor cells by dramatically upregulating production of several factors including the drug transporter P-glycoprotein, the cholesterol binding protein caveolin, and components of lipid raft microdomains of cellular plasma membranes. These raft domains are enriched in cholesterol and sphingolipids and play key roles in signal transduction processes. The distinct composition of plasma membranes of MDR cancers may enable selective chemotherapy targeting these cancers. During the last grant cycle, novel small molecules were synthesized that comprise cholesterylamine covalently linked to protein ligands such as biotin. These compounds bind lipid rafts in plasma membranes of cancer cells. Treatment of cancer cell lines with a synthetic biotin-cholesterylamine ligand (ligand #1 ) and the protein Streptavidin (SA) efficiently targets SA to lipid rafts, resulting in rapid clathrin-mediated endocytosis of this protein-ligand complex. This novel system mimics penetration of cells by Cholera toxin, which binds ganglioside GM1 in lipid rafts. This project is based on the hypothesis that ligand #1 will regulate endocytosis of SA linked to endosome-activated toxins daunorubicin and exotoxin in cancer cell lines. Since ligand #1 binds lipid rafts, selective delivery of SA-linked toxins to lipid raft-rich MDR cancer cells will be investigated in vitro and in vivo in murine cancer models. The effectiveness of ligand #1 at enhancing endocytosis of SA- toxins fused to neuropeptide Y, which targets specific receptors on neuroblastoma cells will also be evaluated. This novel approach directed at enhancing endocytosis of surface receptors by targeting to lipid rafts with small molecules could address the major problem in immunotherapy of non-internalized tumor antigens. The hypothesis that ligand-regulated delivery of SA to antigen presenting cells (APCs) will stimulate immune responses will also be tested. Novel immunostimulants will be investigated by fusing SA to the ovalbumin antigen, regulating endocytosis in APCs with ligand #1, and analyzing T-cell activation. This approach could control immunostimulation at the molecular level and yield novel tools for vaccine development. Recruitment of intracellular avidin fusion proteins to plasma membranes by ligand #1 will also be studied in an effort to conditionally regulate cellular growth and death.

多重耐药（MDR）癌症仍然是治愈性癌症化疗的主要障碍。多重耐药癌细胞与典型肿瘤细胞的不同之处在于显着上调多种因子的产生，包括药物转运蛋白 P-糖蛋白、胆固醇结合蛋白小窝蛋白和脂筏成分细胞质膜的微区。这些筏结构域富含胆固醇和鞘脂在信号转导过程中发挥关键作用。血浆的独特成分耐多药癌症的细胞膜可能能够针对这些癌症进行选择性化疗。最后期间格兰特循环中，合成了新的小分子，其中包含共价连接的胆固醇胺蛋白质配体，例如生物素。这些化合物结合癌细胞质膜中的脂筏。用合成生物素-胆固醇胺配体（配体#1）和蛋白质处理癌细胞系链霉亲和素 (SA) 有效地将 SA 靶向脂筏，从而导致网格蛋白介导的快速内吞作用蛋白质-配体复合物。这种新颖的系统模拟霍乱毒素渗透细胞，该毒素结合脂筏中的神经节苷脂 GM1。该项目基于配体 #1 将调节内吞作用的假设 SA 与癌细胞系中内体激活的毒素柔红霉素和外毒素有关。由于配体#1 结合脂筏，将 SA 连接的毒素选择性递送至富含脂筏的 MDR 癌细胞将在以下研究中进行研究小鼠癌症模型的体外和体内。配体#1在增强SA-内吞作用方面的有效性与神经肽 Y 融合的毒素也将靶向神经母细胞瘤细胞上的特定受体评价。这种新方法旨在通过靶向脂质来增强表面受体的内吞作用小分子筏可以解决非内化肿瘤免疫治疗的主要问题抗原。配体调节的 SA 递送至抗原呈递细胞 (APC) 的假设刺激免疫反应也将受到测试。将通过将 SA 融合来研究新型免疫刺激剂卵清蛋白抗原，用配体 #1 调节 APC 的内吞作用，并分析 T 细胞激活。这该方法可以在分子水平上控制免疫刺激并产生新的疫苗工具发展。配体 #1 将细胞内亲和素融合蛋白招募到质膜上也将进行研究以有条件地调节细胞生长和死亡。