Combination of TNF-Gold Nanoparticles with Radiation

TNF-金纳米粒子与辐射的组合

• 批准号: 7592958
• 负责人: Jacek Capala
• 金额: $ 10.96万
• 依托单位: DIVISION OF BASIC SCIENCES - NCI
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/7592958
• 关键词: Animals; Area; Arthritis; Biodistribution; Biological Assay; Blood Vessels; Breast Cancer Cell; CCR; Caliber; Cancer cell line; Cell Line; Cell Survival; Cells; Clinic; Clinical; Collaborations; Computer Simulation; Condition; Data; Deposition; Detection; Disease; Dose; Drug Delivery Systems; Electron Microscopy; Electrons; Elements; Environmental Impact; External Beam Radiation Therapy; Gold; Gold Colloid; Hour; IACUC; In Vitro; Incubated; Injection of therapeutic agent; Kidney; Laboratories; Lead; Link; Liver; Lung; MCF7 cell; Mass Spectrum Analysis; Measurable; Measures; Medicine; Methodology; Modeling; Monitor; Mus; Myomatous neoplasm; Normal tissue morphology; Nucleotides; Pancreas; Patients; Pharmaceutical Preparations; Phase I Clinical Trials; Production; Property; Proteins; Protocols documentation; Radiation; Radiation Physics; Range; Relative (related person); Research; Research Design; Scanning; Science; Series; Skin; Spleen; Standards of Weights and Measures; Stomach; TNF gene; Techniques; Therapeutic; Time; Tissues; Toxic effect; Translations; Tumor Tissue; Validation; X-Ray Computed Tomography; Xenograft procedure; absorption; bone; clinical application; density; gold chloride; in vivo; infancy; interest; irradiation; nanocarrier; nanomedicine; nanoparticle; nanoscale; neoplastic cell; novel; particle; radiation effect; research study; response; simulation; size; sodium citrate; software development; subcutaneous; synthetic peptide; tool; trafficking; treatment planning; tumor

Background and Significance Nanoparticles (particles < 100 nm) have generated increasing interest in the field of medicine as tools for disease detection, and drug delivery. The field of nanomedicine, while still in its infancy, holds tremendous promise as we begin to understand issues related to the benefits, toxicity and environmental impact of nanoscale materials. Colloidal gold is a neutral gold particle synthesized through the combination of gold chloride and sodium citrate. It has been used safely for decades as a therapeutic for patients with arthritis. The particle measures 20-30 nm in diameter and can be linked irreversibly to proteins, peptides, synthetic drugs and nucleotides. In addition to its properties as a nano-carrier, colloidal gold also acts as a high-Z element. Theoretically, the irradiation of high-Z elements at their K-edge absorption energy leads to emission of Auger electrons and photoelectrons, releasing a large amount of energy at their immediate vicinity. This secondary radiation will result in extra damage to tumor cells at the same dose of the applied external radiation. We believe that this secondary property of colloidal gold opens a significant area of novel research and clinical application. Specifically, because of the preferential trafficking of the CYT-6091 particles to tumor tissue, we can utilize this property to enhance the efficacy of external beam irradiation by increasing the radiation dose deposited into the tumor while minimizing toxicity to normal tissues. The fact that CYT-6091 is currently being evaluated as single agent therapy in the context of a Phase I clinical trial within the CCR, makes the potential for the translation of our findings combining CYT-6091 with radiation in the laboratory to the clinic significant. Research Design Aim 1. In order to identify irradiation conditions optimal for production of secondary radiation in gold particles and to estimate the increase of the radiation dose due to the presence of gold nanoparticles, radiation transport calculations will be carried out using the ESG4 Monte Carlo simulation package or the treatment planning software developed within the ROB Radiation Physics Section. The changes of the radiation dose deposited in tissues as a function of the concentration of gold and the quality of the applied external irradiation will be calculated using standard methodology This data will allow us to determine the relative benefit of the addition of colloidal gold as well as the best concentrations of colloidal gold and the optimal energy of radiation to be used in order to optimize the enhanced radiation effects. Aim 2. Response of tumor cells to combination of pegylated colloidal gold TNF nanoparticles with various types of radiation will be characterized by the clonogenic survival assay. Human breast cancer cell lines, MCF7 and MDA251, with different sensitivities to TNF, will be incubated with or without gold nanoparticles and TNF, and exposed to different doses (0 8 Gy) of radiation. The energy of applied x-rays will vary in the range from 5 keV to 60 MeV. The effects of radiation on cell survival will be quantified and evaluated. This series of experiments will help in the validation of the predicted doses from Aim 1 and will allow for the selection of the appropriate cell line, and radiation type and dose for in vivo studies. Aim 3. Initially, CT scans of a tissue-equivalent phantom with inserts containing different concentrations of gold will be used to assess the changes of density of tissue due to the presence of gold nanoparticles and the feasibility of using CT scans to monitor their in vivo biodistribution. Results of the CT scans will be compared to the quantification of the particles in the tissue by validated electron microscopy approaches. If the results of these studies are positive, the micro-CT scanner will be used to compare the changes of Hounsfield units on the CT scans with gold concentration measured in the tissue. Mice with measurable subcutaneous tumors (approximately 0.5 cm3) will be injected with CYT-6091 at doses that may lead to concentrations in the tumors identified as effective by the computer simulations and the in vitro studies. Three and five hours post injection, CT scans will be carried out. Groups of five mice per time point will be used. After completion of the scans, the mice will be euthanized and tissues of interest including tumor, muscle, lung, liver, spleen, pancreas, kidney, stomach, skin, and bone will be collected. All mouse studies will be performed under approved IACUC protocols. The concentration of gold in the tissues will be measured using mass spectrometry techniques. Electron microscopy will be used to assess the sub-cellular microdistribution of gold. Aim 4. The possible synergistic effect of combining x-rays with CYT-6091 will be studied in vivo using both mouse xenograft and syngeneic tumor models. Groups of 10 tumor-bearing animals will be treated with: 1) radiation only; 2) pegylated colloidal gold TNF only; and 3) combination radiation plus pegylated colloidal gold TNF. Non-treated animals will be used as controls. Applied doses of the nanoparticles and radiation will be identified in previous Aims. Tumor size and survival will be used to assess the efficacy of each treatment. Project Status: 1. In vitro clonogenic survival experiments have been carried out to establish the effect of nanoparticles in combination with radiation on cell survival. 2. Preliminary in vivo studies using tumor bearing mice are carried out

背景和意义纳米颗粒（颗粒< 100 nm）已经产生 作为疾病检测工具的医学领域的兴趣日益增加， 交付.纳米医学领域，虽然仍处于起步阶段，拥有巨大的希望， 我们开始了解有关的利益，毒性和环境影响的问题， 纳米材料胶体金是一种中性的金颗粒， 氯化金和柠檬酸钠的组合。几十年来，它一直被安全地用作 关节炎患者的治疗药物。该颗粒的直径为20-30 nm， 与蛋白质、肽、合成药物和核苷酸不可逆地连接。除了其 除了作为纳米载体的特性外，胶体金还充当高Z元素。理论上 高Z元素在其K边吸收能处的辐照导致俄歇发射 电子和光电子，在它们的附近释放大量的能量。 这种二次辐射将导致在相同剂量下对肿瘤细胞的额外损伤。 应用外部辐射。我们认为胶体金的这种次要性质开启了一种 新研究和临床应用的重要领域。具体来说，由于 CYT-6091颗粒向肿瘤组织的优先运输，我们可以利用这一点， 通过增加辐射剂量来增强外部射束辐射的功效的特性 沉积到肿瘤中，同时最小化对正常组织的毒性。CYT-6091 目前正在I期临床试验中作为单药治疗进行评估 CCR内的试验，使我们的研究结果结合翻译的潜力 CYT-6091在实验室用放射线对临床有显著意义。研究设计目标1.在 为了确定在金中产生二次辐射的最佳辐照条件， 粒子，并估计由于金的存在而增加的辐射剂量 纳米粒子，辐射输运计算将使用ESG 4 Monte Carlo 模拟包或在ROB辐射内开发的治疗计划软件 物理科。沉积在组织中的辐射剂量的变化作为一个函数， 将计算金的浓度和所施加的外部照射的质量 使用标准方法，这些数据将使我们能够确定 胶体金的添加量以及胶体金和 为了优化增强的辐射效果，需要使用最佳的辐射能量。 目标2.肿瘤细胞对聚乙二醇化胶体金TNF纳米粒联合应用的反应 将通过克隆形成存活测定来表征各种类型的辐射。人类 乳腺癌细胞系，MCF 7和MDA 251，具有不同的敏感性，TNF， 与金纳米颗粒和TNF一起孵育或不孵育，并暴露于不同剂量（0 8戈伊）。所施加的X射线的能量将在从5 keV至20 keV的范围内变化。 60 MeV将对辐射对细胞存活的影响进行量化和评价。这 一系列实验将有助于验证目标1的预测剂量， 允许选择适当的细胞系，以及用于体内的辐射类型和剂量。 问题研究目标3.最初，CT扫描的组织等效体模的插入物包含 不同浓度的金将用于评估组织密度的变化， 金纳米粒子的存在以及使用CT扫描来监测其 体内生物分布。CT扫描的结果将与 通过经验证的电子显微镜方法检测组织中的颗粒。如果这些结果 研究是积极的，将使用微型CT扫描仪来比较Hounsfield的变化 单位的CT扫描与金浓度测量的组织。具有可测量的 皮下肿瘤（约0.5cm3）将注射CYT-6091，剂量可 导致肿瘤中的浓度被计算机模拟识别为有效的， 体外研究。注射后3小时和5小时，将进行CT扫描。 每个时间点将使用5只小鼠的组。完成扫描后，小鼠将 将其安乐死，并将感兴趣的组织包括肿瘤、肌肉、肺、肝、脾、胰腺， 收集肾脏、胃、皮肤和骨骼。所有小鼠研究将在 批准的IACUC方案。组织中的金浓度将使用质量 光谱技术。电子显微镜将用于评估亚细胞 黄金的微观分布目标4。X射线和放射性物质结合的可能的协同效应 CYT-6091将使用小鼠异种移植物和同基因肿瘤模型进行体内研究。组 10只荷瘤动物将接受以下治疗：1）仅放射治疗; 2）聚乙二醇化胶体 仅金TNF;和3）组合辐射加聚乙二醇化胶体金TNF。未处理 动物将用作对照。纳米粒子和辐射的应用剂量将是 在过去的目标中。肿瘤大小和存活率将用于评估 每一次治疗。项目状态：1.进行了体外克隆存活实验 研究人员试图确定纳米粒子与辐射结合对细胞存活的影响。 2.使用荷瘤小鼠进行初步体内研究