Self-Assemblying Immunotherapeutic Nanorings

自组装免疫治疗纳米环

• 批准号: 8055479
• 负责人: CARSTON R. WAGNER
• 金额: $ 29.36万
• 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 2013-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8055479
• 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; Goals; 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/lymphoma; 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用作生物传感器和治疗剂的纳米版本的设计和开发，对许多人类疾病的治疗将大大提高。由于它们已经被用来向癌症组织和免疫细胞提供多种毒素，药物和放射性核素，因此可以将单克隆和重组抗体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，放射性核素和毒素对肿瘤和免疫细胞的靶向已被证明是成功的成像和治疗方法。尽管它们的临床前和临床成功，但单克隆抗体放射性核素的发展遭受了许多担忧，例如由于肾脏清除率低，毒性放射性核素骨沉积，毒性放射性核素骨沉积，结合化学性能与免疫反应性和分子异质性不符。我们建议开发一种由药理学控制的组装和拆卸的多价放射性疗法纳米结构的方案，该纳米结构有可能纳入BI特异性。我们将利用最近发现如何从大肠杆菌二氢酸酯还原酶（DHFR-DHFR）融合蛋白中构建离散化学诱导的蛋白纳米（8-30 nm DIA。）的优势。我们将制备由Daniel Vallera博士（共同入侵器）开发的单链抗体（SCFV）融合的DHFR-DHFR分子，并与正常的B细胞和B细胞淋巴瘤和白血病抗原CD19结合。我们将准备DHFR-DHFR-DHFR-ANTI-CD19 SCFV融合蛋白，这些蛋白能够在存在与荧光小子或螯合放射性核素的甲氨蝶呤二聚体的情况下自组装成二价，四位或八值物种。我们将证明抗体纳入能够在体外结合并经历细胞内B-白血病细胞的摄取和运输。此外，我们还将确定抗体纳米菌的体内生物分布，以及Timethoprim（一种无毒的大肠杆菌DHFR抑制剂）促进体内低聚物拆卸的能力。我们还将研究使用小鼠异种移植肿瘤模型的DHFR-DHFR-抗CD19 SCFV纳米模型的抗肿瘤和肿瘤成像特性。尽管我们将重点介绍用于治疗B细胞癌和自身免疫性疾病的抗体 - 降低核素纳米含量的特定设计，但本研究阐明的原理将适用于设计和治疗广泛疾病的治疗性纳米剂。 公共卫生相关性：可以在基于疾病的组织中回家的纳米颗粒的发展，报告组织的位置并破坏组织是我们研究的目标。在我们的第一次尝试中，我们将开发一种方法来制备可以靶向B细胞和B细胞白血病的放射性标记抗体蛋白纳米纳米纳米纳米。我们将对肿瘤和免疫细胞成像和抗肿瘤疗法使用这些抗体纳米疗法，并证明我们可以在需要时去除纳米颗粒。