Self-Assemblying Immunotherapeutic Nanorings

自组装免疫治疗纳米环

• 批准号: 7649435
• 负责人: CARSTON R. WAGNER
• 金额: $ 30.07万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-07-02 至 2012-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7649435
• 关键词: Acids; Address; Aftercare; Amines; Antibodies; Area; Autoimmune Diseases; B-Cell Lymphomas; B-Lymphocytes; Back; Binding; Binding Sites; Biodistribution; Biological; Biosensor; Blood Circulation; CD19 Antigens; CD19 gene; Caliber; Cell surface; Cells; Chelating Agents; Chemistry; Chimeric Proteins; Chronic Lymphocytic Leukemia; Clinical; Coupled; DNA; Deposition; Detection; Development; Diagnostic; Dihydrofolate Reductase; Dihydrofolate Reductase Inhibitor; Disease; Drug Delivery Systems; Enzyme Inhibitor Drugs; Enzyme Inhibitors; Escherichia coli; Face; Figs - dietary; Goals; Half-Life; Heterogeneity; Histocompatibility Testing; Home environment; Image; Immune; Immunotherapeutic agent; In Vitro; Laboratories; Length; Ligands; Malignant Neoplasms; Methods; Methotrexate; Modeling; Molecular; Monoclonal Antibodies; Mus; Nanostructures; Nucleic Acids; Peptides; Pharmaceutical Preparations; Preparation; Property; Proteins; Protocols documentation; Radioimmunoconjugate; Radioisotopes; Radiolabeled; Recombinant Antibody; Recombinants; Relative (related person); Renal clearance function; Reporting; Research; Research Personnel; Role; Specificity; Testing; Therapeutic; Therapeutic Agents; Tissue Engineering; Tissues; Toxin; Triazines; base; bone; cellular imaging; design; dimer; fluorophore; human disease; immunoreactivity; in vivo; leukemia; monomer; nanodevice; nanomaterials; nanoparticle; novel; pre-clinical; public health relevance; radiotracer; scaffold; success; trafficking; tumor; tumor xenograft; uptake

DESCRIPTION (provided by applicant): The treatment of a number of human diseases would be greatly advanced by the design and development of nanodevices capable of functioning both as biosensors and therapeutic agents. Because they have already been used to deliver a variety of toxins, drugs and radionuclides to cancer tissues and immune cells, monoclonal and recombinant antibodies could be harnessed as recognition ligands for anticancer and immunosuppresive nanostructures. The targeting of radionuclides and toxins to tumors and immune cells by conjugation to antibodies has been shown to be a successful imaging and therapeutic approach. Despite their preclinical and clinical success, the development of monoclonal antibody radionuclide conjugates suffers from a number of concerns, such as, poor imaging capability due to low renal clearance, toxic radionuclide bone deposition, conjugation chemistry that is incompatible with immunoreactivity and molecular heterogeneity. We propose to develop a protocol for the pharmacologically controlled assembly and disassembly of multivalent radioimmunotherapeutic nanostructures that have the potential for incorporating bi-specificity. We will take advantage of our recent discovery of how to construct discrete chemically induced protein nanorings (8-30 nm dia.) from E. coli dihydrofolate reductase (DHFR-DHFR) fusion proteins. We will prepare DHFR-DHFR molecules fused to a single chain antibody (scFv) that was developed by Dr. Daniel Vallera (co-Investigator) and binds to the normal B-cell and B-cell lymphoma and leukemia antigen, CD19. We will prepare DHFR- DHFR-anti-CD19 scFv's fusion proteins that are able to self-assemble into bivalent, tetravalent or octavalent species in the presence of a methotrexate dimerizer coupled to a fluorophore or chelated radionuclides. We will demonstrate that the antibody-nanorings are able to selectively bind and undergo intracellular B-leukemia cell uptake and trafficking in vitro. In addition, we will determine the in vivo biodistribution of the antibody nanorings, as well as the ability of timethoprim, a non-toxic E. coli DHFR inhibitor, to promote oligomer disassembly in vivo. We will also investigate the anti-tumor and tumor imaging properties of the DHFR-DHFR- anti-CD19 scFv nanorings with a mouse xenograft tumor model. Although we will focus on the specific design of antibody-radionuclide nanorings for the treatment of B-cell cancers and autoimmune diseases, the principles elucidated by this study will be applicable to the design of therapeutic nanorings for the detection and treatment of a wide range of diseases. PUBLIC HEALTH RELEVANCE: The development of nanoparticles that can home in on disease based tissues, report back on where the tissue is and destroy the tissue is the goal of our research. In our first attempt, we will develop a method to prepare radiolabeled antibody protein nanorings that can target B-cells and B-cell leukemia's. We will use these antibody-nanorings for both tumor and immune cell imaging and antitumor therapy and demonstrate that we can remove the nanoparticles when needed.

描述（由申请人提供）：通过设计和开发能够同时充当生物传感器和治疗剂的纳米装置，将极大地推进许多人类疾病的治疗。由于单克隆抗体和重组抗体已被用于向癌组织和免疫细胞递送各种毒素、药物和放射性核素，因此可以利用它们作为抗癌和免疫抑制纳米结构的识别配体。通过与抗体结合将放射性核素和毒素靶向肿瘤和免疫细胞已被证明是一种成功的成像和治疗方法。尽管在临床前和临床上取得了成功，单克隆抗体放射性核素缀合物的开发仍面临许多问题，例如由于肾脏清除率低而导致成像能力差、有毒放射性核素骨沉积、与免疫反应性不相容的缀合化学以及分子异质性。我们建议开发一种药理学控制的多价放射免疫治疗纳米结构的组装和拆卸协议，该纳米结构具有合并双特异性的潜力。我们将利用我们最近的发现，即如何从大肠杆菌二氢叶酸还原酶 (DHFR-DHFR) 融合蛋白构建离散的化学诱导蛋白纳米环（直径 8-30 nm）。我们将制备与单链抗体 (scFv) 融合的 DHFR-DHFR 分子，该抗体由 Daniel Vallera 博士（共同研究员）开发，并与正常 B 细胞和 B 细胞淋巴瘤和白血病抗原 CD19 结合。我们将制备 DHFR-DHFR-抗 CD19 scFv 融合蛋白，该融合蛋白能够在与荧光团或螯合放射性核素偶联的甲氨蝶呤二聚体存在的情况下自组装成二价、四价或八价种类。我们将证明抗体纳米环能够选择性结合并在体外进行细胞内 B 白血病细胞的摄取和运输。此外，我们将确定抗体纳米环的体内生物分布，以及无毒大肠杆菌DHFR抑制剂timethoprim促进体内低聚物分解的能力。我们还将通过小鼠异种移植肿瘤模型研究 DHFR-DHFR-抗 CD19 scFv 纳米环的抗肿瘤和肿瘤成像特性。虽然我们将重点关注用于治疗 B 细胞癌症和自身免疫性疾病的抗体-放射性核素纳米环的具体设计，但本研究阐明的原理将适用于用于检测和治疗多种疾病的治疗性纳米环的设计。 公共健康相关性：我们研究的目标是开发能够追踪疾病组织、报告组织位置并破坏组织的纳米颗粒。在我们的第一次尝试中，我们将开发一种方法来制备可靶向 B 细胞和 B 细胞白血病的放射性标记抗体蛋白纳米环。我们将使用这些抗体纳米环进行肿瘤和免疫细胞成像以及抗肿瘤治疗，并证明我们可以在需要时去除纳米颗粒。