Multivalent Nanophage Engineered as Dual Receptor Cancer Theranostic Agents.

多价纳米噬菌体被设计为双受体癌症治疗剂。

• 批准号: 8569062
• 负责人: SUSAN L DEUTSCHER
• 金额: $ 16.6万
• 依托单位: UNIVERSITY OF MISSOURI-COLUMBIA
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-07-01 至 2015-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8569062
• 关键词: Acids; Address; Affinity; Antibodies; Antigen Targeting; Antigens; Avidity; Bacteriophages; Beta Particle; Binding; Biodistribution; Biological; Biomedical Engineering; Biotin; Bispecific Antibodies; Blood; Breast Carcinoma; Cancer Detection; Capsid Proteins; Carcinoma; Cell Line; Cells; Combined Modality Therapy; Coupled; DOTA-biotin; Detection; Development; Diagnosis; Disease; Dissociation; Dose; Dose-Limiting; Drug Kinetics; Engineering; Event; Generations; Genetic Engineering; Goals; Hour; Human; Image; In Vitro; Label; Length; Lutetium; Malignant Epithelial Cell; Malignant Neoplasms; Malignant neoplasm of prostate; Multimodal Imaging; Mus; Neoplasm Metastasis; Normal tissue morphology; Nude Mice; Optics; Peptides; Phage Display; Play; Process; Production; Property; Public Health; Radioisotopes; Receptor Signaling; Research; Resistance; Signal Transduction; Silicon Dioxide; Specificity; Staging; Streptavidin; Therapeutic; TimeLine; Tissues; Translating; Tumor Antigens; Virion; Work; Xenograft procedure; antigen binding; base; cancer cell; cancer therapy; cyclen; dimer; immunogenicity; improved; in vivo; malignant breast neoplasm; nanometer; nanoparticle; novel; optical imaging; particle; public health relevance; receptor; research study; scaffold; single photon emission computed tomography; theranostics; tumor; tumor progression; tumorigenesis

DESCRIPTION (provided by applicant): Nearly all cancers are treatable in their earliest forms, but often fatal once metastasis has occurred. Improved diagnosis and therapy of cancer will result from a more directed approach in which antigens specific to or over expressed on tumors are targeted. Although therapies targeted to a single antigen have proven moderately successful, poor specificity and affinity, coupled with resistance have limited their use. The goal of this research is to develop a bacteriophage (phage)-based platform for targeting multiple cancer antigens regardless of spatial or biological relatedness. The advantage to dual targeting includes both increased avidity through decreased simultaneous dissociation rates, and increased selectivity through multiple antigen binding. Numerous receptors have been shown to play key roles in tumorigenesis but have proven difficult to target. For example, the ErbB2 and ErbB3 receptors are both present in early stage and resistant carcinomas, but absent or present at low levels in normal tissues. Although the receptors can heterodimerize, the majority do not and are not close enough for targeting by current heterovalent platforms, such as bispecific antibody constructs. Phage provide a scaffold that spans hundreds of nanometers and display multiple copies of targeting peptides, which can be utilized to bind antigens regardless of interaction or separation. It is hypothesized that phage displaying targeted peptides at opposite tips of the virion can simultaneously bind multiple cancer antigens (such as Erb2 and Erb3) regardless of their proximity or interaction status. Such heteromultivalent (HMV) phage, engineered to appropriate lengths, will be employed in a novel, amplified 3-step pretargeting approach. Truncated, biotinylated HMV phage will be injected for in vivo tumor targeting. Following phage clearance, fluorescent streptavidin-coated silica nanoparticles will be used to target the multiple biotins on tumor-bound phage. The nanoparticles have demonstrated rapid blood clearance and will allow for multimodal imaging and a second level of signal amplification. Biotin conjugated to the 1,4,7,10- tetraazacyclododecane-1,4,7,10-tetraacetic acid (DOTA)-chelated radionuclide [177Lu] lutetium will deliver high levels of a theranostic agent that can be simultaneously used for single photon emission computed tomography (SPECT) imaging in addition to providing a high dose of tumor-localized beta radiation. The specific aims include: 1) genetically engineer heteromultivalent (HMV) nanophage of discrete lengths (50-750 nm) displaying ErbB2 and ErbB3 targeting peptides, 2) characterize HMV nanophage affinity, specificity, and avidity to ErbB2/ErbB3, and 3) characterize the tumor targeting and pharmacokinetics of the nanophage using the amplified three-step pretargeting approach in ErbB2+/ErbB3+ BT-474 human breast cancer xenografted mice. These phage will yield a new imaging and therapeutic platform with high affinity, selectivity and optimum pharmacokinetics. Furthermore, the phage can be engineered to display targeting peptides or antibodies for any antigen(s) of choice, providing great versatility as a novel theranostic agent.

描述（由申请人提供）：几乎所有癌症在其早期形式下都是可以治疗的，但一旦发生转移通常是致命的。癌症诊断和治疗的改进将源自更直接的方法，其中针对肿瘤特异或过度表达的抗原。尽管针对单一抗原的疗法已被证明取得了一定的成功，但特异性和亲和力较差，再加上耐药性，限制了它们的使用。目标 这项研究的目的是开发一个基于噬菌体（噬菌体）的平台，用于靶向多种癌症抗原，无论空间或生物相关性如何。双重靶向的优点包括通过降低同时解离速率来增加亲和力，以及通过多种抗原结合来增加选择性。许多受体已被证明在肿瘤发生中发挥关键作用，但已被证明难以靶向。例如，ErbB2和ErbB3受体均存在于早期癌和耐药癌中，但在正常组织中不存在或以低水平存在。尽管受体可以异二聚化，但大多数受体不能异二聚化，并且距离不够近，无法被当前的异价平台（例如双特异性抗体构建体）靶向。噬菌体提供了一个跨越数百纳米的支架，并展示多个拷贝的靶向肽，这些肽可用于结合抗原，无论相互作用或分离如何。据推测，在病毒颗粒相对尖端展示靶向肽的噬菌体可以同时结合多种癌症抗原（例如 Erb2 和 Erb3），无论它们的接近程度或相互作用状态如何。这种异多价 (HMV) 噬菌体被设计成适当的长度，将用于新型的扩增三步预靶向方法。将注射截短的生物素化 HMV 噬菌体以进行体内肿瘤靶向。噬菌体清除后，荧光链霉亲和素包被的二氧化硅纳米粒子将用于靶向肿瘤结合噬菌体上的多种生物素。纳米粒子已被证明可以快速血液清除，并将允许多模态成像和第二级信号放大。与 1,4,7,10-四氮杂环十二烷-1,4,7,10-四乙酸 (DOTA) 螯合放射性核素 [177Lu] 镥结合的生物素将提供高水平的治疗诊断剂，除了提供高剂量的肿瘤局部 β 之外，还可同时用于单光子发射计算机断层扫描 (SPECT) 成像 辐射。具体目标包括：1) 对离散长度 (50-750 nm) 的异多价 (HMV) 纳米噬菌体进行基因改造，展示 ErbB2 和 ErbB3 靶向肽，2) 表征 HMV 纳米噬菌体对 ErbB2/ErbB3 的亲和力、特异性和亲和力，以及 3) 使用扩增的纳米噬菌体的肿瘤靶向和药代动力学表征 ErbB2+/ErbB3+ BT-474 人乳腺癌异种移植小鼠的三步预靶向方法。这些噬菌体将产生具有高亲和力、选择性和最佳药代动力学的新成像和治疗平台。此外，噬菌体可以被设计为展示针对任何所选抗原的靶向肽或抗体，从而提供作为新型治疗诊断剂的强大多功能性。