Dual-pronged nano-drug delivery using plant virus-like particles

使用植物病毒样颗粒的双管齐下纳米药物输送

• 批准号: 9372245
• 负责人: Nicole Franziska Steinmetz
• 金额: $ 47.01万
• 依托单位: CASE WESTERN RESERVE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-08-01 至 2018-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9372245
• 关键词: 4T1; Address; Anatomy; Animals; Antibodies; Applications Grants; Artificial nanoparticles; Biological; Breast Cancer Patient; Capsid Proteins; Cells; Clinical; Colon Carcinoma; Cowpea Mosaic Viruses; Data; Development; Disease; Distant Metastasis; Doxorubicin; Drug Delivery Systems; ERBB2 gene; Estrogen Receptors; Flow Cytometry; Histocytochemistry; Image; Immune; Immune response; Immunofluorescence Immunologic; Immunologic Memory; Immunotherapy; Knockout Mice; Lead; Libraries; Life; Ligands; Location; Malignant Neoplasms; Malignant neoplasm of ovary; Mediating; Memory; Molecular; Mosaic Viruses; Mouse Strains; Mus; Nanotechnology; Nature; Neoplasm Metastasis; Nucleic Acids; Oncolytic; Particulate; Pathway interactions; Patients; Pharmaceutical Preparations; Plant Viruses; Plants; Play; Primary Neoplasm; Progesterone Receptors; Recurrence; Research; Role; Shapes; Signal Pathway; Signal Transduction; TLR7 gene; Testing; Therapeutic; Time; Toll-like receptors; Translating; Treatment Efficacy; Tumor Debulking; Tumor Immunity; Virus; Virus-like particle; Woman; antitumor effect; arm; base; chemotherapy; engineering design; immune activation; immunogenic; improved; in vivo imaging; insight; melanoma; mortality; mouse model; nanocarrier; nanodrug; nanoparticle; nanotherapy; outcome forecast; particle; response; success; targeted treatment; therapy outcome; triple-negative invasive breast carcinoma; tumor; tumor microenvironment

Summary Triple negative breast cancer (TNBC) is an aggressive life-threatening disease with poor prognosis and limited treatment options. To address this clinical need, we propose a dual-pronged drug delivery approach that combines chemo- and immune-drugs with a highly potent drug delivery platform, that itself has anti-tumor efficacy. This grant application is based on our recent demonstration that virus-like particles (VLP) from the plant virus cowpea mosaic virus (CPMV), when introduced into the tumor microenvironment, elicit potent local and systemic anti-tumor immunity and protects animals from outgrowth of metastatic disease or recurrence of the disease [Nature Nanotechnology 2016]. We have demonstrated efficacy in multiple mouse models treating primary and metastatic tumors in multiple anatomic locations and multiple mouse strains, including mouse models of TNBC. The nanoparticle engineering design space and pathway of immune-activation is not yet un- derstood, and one arm of this R01 grant application will decipher the underlying mechanism. Second, we will develop and test dual-pronged therapeutic approaches through combination of the nanoparticle platform with chemotherapies and immune-drugs to potentiate the therapy and enable treatment of late-stage disease. We will fulfill the following specific aims: 1) We will address whether the anti-tumor effect is CPMV-specific, wheth- er the particulate, multivalent nature is a requirement, and whether nanoparticle shape and size plays a role. A library of plant-derived VLPs of distinct molecular composition, size, and shape will be studied and efficacy will be assessed using the 4T1 mouse model of TNBC. 2) We will determine the cells and signaling pathways in- volved in the VLP-induced anti-tumor immune response. Longitudinal imaging in combination with histochemis- try and flow cytometry will provide comprehensive insight into the biological response. To identify the potential signaling pathways involved, studies will be performed in knockout mouse models that lack ASC, Myd88 or Trif adaptor molecules. 3) Immunotherapy as monotherapy is generally effective only against small tumors, and most patients do not respond to single approach immunotherapy. The combination of therapeutic regimes, i.e. the combination of multiple immunotherapies or combination with chemotherapy, however, could form the ba- sis for success. First, we will combine VLPs with doxorubicin (an immunogenic chemotherapy) through co- administration (unconjugated) or co-delivery (conjugated to the VLP). Second, we will combine VLPs with known `immunogenic danger signals' and co-deliver ssRNA or CpGs to activate signaling through Toll-like re- ceptors. We hypothesize that the combination of the immune-stimulatory VLP and chemo- or immune-drugs will potentiate efficacy while mounting a systemic anti-tumor immune response and memory, and this will be tested through treatment of mice bearing 4T1 primary and metastatic tumors. The recent approval of the onco- lytic immunotherapy T-VEC highlights the potential of virus-based therapeutics. Our application is distinct, be- cause we use a plant VLP for dual-pronged tumor treatment and priming of anti-tumor immune memory.

概括 三阴性乳腺癌（TNBC）是一种侵袭性危及生命的疾病，预后不良， 治疗选择有限。为了满足这一临床需求，我们提出了一种双管齐下的药物输送方法 将化疗药物和免疫药物与高效的药物输送平台相结合，该平台本身具有抗肿瘤作用 功效。这项拨款申请是基于我们最近的证明，即病毒样颗粒（VLP）来自 植物病毒豇豆花叶病毒（CPMV），当被引入肿瘤微环境时，引发有效的局部 和全身抗肿瘤免疫力，并保护动物免受转移性疾病的发展或复发 这种疾病[自然纳米技术2016]。我们已经在多个小鼠模型中证明了治疗的功效 多个解剖位置和多种小鼠品系的原发性和转移性肿瘤，包括小鼠 TNBC 的模型。纳米颗粒工程设计空间和免疫激活途径尚未确定 明白了，这个 R01 拨款申请的一个分支将破译底层机制。其次，我们将 通过将纳米颗粒平台与 化疗和免疫药物可增强治疗效果并能够治疗晚期疾病。我们 将实现以下具体目标：1）我们将解决抗肿瘤作用是否是 CPMV 特异性的，是否- 对于颗粒，多价性质是一个要求，并且纳米颗粒的形状和尺寸是否起作用。一个 将研究具有不同分子组成、大小和形状的植物源 VLP 文库，并研究功效 使用 TNBC 的 4T1 小鼠模型进行评估。 2）我们将确定以下细胞和信号通路： 参与VLP诱导的抗肿瘤免疫反应。纵向成像与组织化学相结合 尝试和流式细胞术将提供对生物反应的全面了解。识别潜力 所涉及的信号通路，研究将在缺乏 ASC、Myd88 或 Trif 的敲除小鼠模型中进行 接头分子。 3) 免疫疗法作为单一疗法通常仅对小肿瘤有效，并且 大多数患者对单一方法免疫疗法没有反应。治疗方案的组合，即 然而，多种免疫疗法的组合或与化疗的组合可以形成ba- 姐姐为了成功。首先，我们将 VLP 与阿霉素（一种免疫原性化疗）通过共 给药（未结合）或共同递送（与 VLP 结合）。其次，我们将 VLP 与 已知的“免疫原性危险信号”并共同传递 ssRNA 或 CpG，以通过 Toll 样重新激活信号传导 受体。我们假设免疫刺激性 VLP 与化疗或免疫药物的组合 将增强疗效，同时建立系统性抗肿瘤免疫反应和记忆，这将是 通过治疗携带 4T1 原发性和转移性肿瘤的小鼠进行测试。最近批准的肿瘤 裂解性免疫疗法 T-VEC 凸显了基于病毒的疗法的潜力。我们的应用程序是独特的，是 因为我们使用植物 VLP 进行双管齐下的肿瘤治疗和启动抗肿瘤免疫记忆。