Site-directed Chemotherapy for Breast Cancer using Novel Angiogenesis Inhibitor

使用新型血管生成抑制剂进行乳腺癌定点化疗

• 批准号: 7660596
• 负责人: SHAKER A MOUSA
• 金额: $ 16.94万
• 依托单位: ALBANY COLLEGE OF PHARMACY
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-08-07 至 2011-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7660596
• 关键词: Adverse effects; Angiogenesis Inhibitors; Area; Binding; Biological Assay; Breast Cancer Cell; Breast Cancer Model; Cancer Patient; Cell Proliferation; Cell membrane; Characteristics; Data; Dermal; Development; Drug Delivery Systems; Drug Formulations; Drug resistance; Effectiveness; Encapsulated; Endothelial Cells; Female; Fluorescein; Fluoresceins; Fluorescence Microscopy; Future; Genomics; Goals; Grant; Growth Factor; Heart Diseases; Hormone Antagonists; Human; Implant; In Vitro; Integrins; Investigation; Knock-in Mouse; Laboratories; Ligands; Link; MCF7 cell; Malignant Neoplasms; Mammary gland; Mediating; Metabolic; Methods; Modeling; Mus; Mutate; Mutation; Neurologic; Nude Mice; Organ; Ovum; Paclitaxel; Pathway interactions; Patients; Performance; Pharmaceutical Preparations; Physiological; Play; Process; Property; Research; Resistance development; Role; Signal Pathway; Site; Stress; Surface; System; Testing; Therapeutic; Thyroid Hormone Receptor; Thyroid Hormones; Thyroid preparation; Thyronines; Thyroxine; Toxic effect; Tumor Angiogenesis; angiogenesis; antiangiogenesis therapy; cancer cell; cancer therapy; carcinogenesis; cell motility; chemotherapeutic agent; chemotherapy; chorioallantoic membrane; clinical application; controlled release; covalent bond; drug testing; hormone analog; improved; in vivo; malignant breast neoplasm; matrigel; membrane model; mouse model; nanoparticle; neoplastic cell; neovascularization; new technology; novel; particle; prevent; public health relevance; receptor; research study; response; tetraiodothyroacetic acid; thyronine; tool; tumor; tumor growth

DESCRIPTION (provided by applicant): Recent evidence has been provided that thyroid hormones may also play a role in carcinogenesis, and pioneering investigations in this area have demonstrated that the thyroid hormones such as 3,5,3'-triiodo-L-thyronine (T3) and L-Thyroxine (T4) can stimulate cancer cell proliferation and thus act as growth factors. We have shown that both T3 and T4 exert potent pro-angiogenic effects in the human dermal microvascular endothelial cell assays and in the chick chorioallantoic membrane (CAM) and Matrigel model of angiogenesis. Taken together, these findings suggest that antagonist for thyroid hormones and/or related signaling pathways may represent compelling targets to treat cancer. Recent studies from our laboratory have shown that tetraiodothyroacetic acid (tetrac), a deaminated thyroid hormone analog, capable of preventing the binding of T4 to 1vB3 at the cell membrane exerted profound inhibitory effects on cellular proliferation and angiogenesis. The finding that tetrac inhibits cancer cell proliferation prompted us to investigate whether it may also stimulate cellular response to stress and thus reverse the development of drug resistance. Our results demonstrate that tetrac impacts on drug resistance and implicate at least two pathways that play key roles in cellular response to chemotherapy. In this Exploratory Application we will investigate the potential use of tetrac as a therapeutic tool for the treatment of cancer, particularly with regards to its possible role in suppressing the development of resistance to chemotherapy. The goal is to determine whether the anti-angiogenic and anti-proliferative actions of tetrac are initiated at the plasma membrane and not at the genomic level. To this end, we have generated PLGA nanoparticles covalently - linked to Tetrac on their surfaces. Preliminary data have shown that upon incubation with cancer cells, particles are preferentially localized to the plasma membrane. Hypothesis: Preferential targeting of Tetrac to the plasma membrane using Tetrac covalently-linked to nanoparticles (TclNPs) will provide a unique approach to study plasma membrane receptor-mediated actions of this hormone antagonist. In the context of the proposed study, we will investigate its anti- angiogenic and anti-proliferative actions. Because thyroid agents used systemically could potentially impact on multiple physiological pathways, the selective targeting of Tetrac to receptor(s) at the plasma membrane may be sufficient to significantly inhibit proliferation of aggressive tumors with fewer side effects. The use of biodegradable TclNPs loaded with a chemotherapeutic agent Paclitaxel can also be used for a controlled release of this drug at the tumor site and thus, is expected to result in better anti-tumor activity with reduced Paclitaxel toxicity. Specific Aims: 1. Experiments will be performed to evaluate the functional activities of TclNPs in vitro in MCF7 cells and in endothelial cells. Preliminary data using fluorescence microscopy showed that fluorescein-loaded TclNPs localize preferentially at the plasma membrane of the breast cancer MCF7 cells. Further studies will be performed to determine binding characteristics of TclNPs, and their effects on proliferation and cell migration. The effects of TclNPs also will be tested on pathways associated with the development or reversal of drug resistance in tumor cells. The effect of TclNPs will be compared to those of free Tetrac in these systems. 2. Pilot experiments to determine the optimum formulations of TclNPs will be performed in ova in the chick chorioallantoic membrane (CAM) tumor implant model of tumor growth and angiogenesis prior to performance of nanoparticle-targeted treatments of nude mice. The CAM model permits in ova pre-screening for bioactivity while limiting the use of more sentient and costly murine species. Nanoparticle formulations that show optimum anti-tumor and anti-angiogenesis activity in the CAM model will be tested in the orthotopic breast cancer model described in Experimental Methods. 3. Female athymic mice will have drug-resistant MCF7 human breast cancer cells implanted orthotopically into the fourth mammary gland. We will evaluate the effectiveness of targeted nanoparticles TclNPs, in reducing tumor growth and tumor angiogenesis. We will evaluate the efficacy of these formulations in limiting potential toxicities associated with systemic Tetrac administration and determine whether TclNPs, with or without Paclitaxel act to limit the development of drug resistance, as suggested by our preliminary in vitro studies. The novel nanoparticle system to be evaluated in these studies combines the properties of targeting and anti-tumor activities, and because Tetrac, Paclitaxel and PLGA nanoparticle systems are all approved for use in patients, could find potential clinical application in the foreseeable future. PUBLIC HEALTH RELEVANCE: Cancer cells have the unique ability to develop resistance to chemotherapeutic drugs, and so research on ways to reverse this phenomenon would have significant value in the treatment of cancer patients. This project will use a combination of two drugs, one of which impairs the cancer cell's ability to develop drug resistance, in a mouse model of breast cancer. A novel technology, the use of nanoparticles to encapsulate the test drugs, and direct them to tumors will be tested to determine whether these nanoparticles can improve the delivery of drugs and minimize the associated toxicities.

描述（由申请人提供）：最近有证据表明甲状腺激素也可能在致癌作用中发挥作用，该领域的开创性研究表明，甲状腺激素如3，5，3 '-三碘-L-甲状腺原氨酸（T3）和L-甲状腺素（T4）可以刺激癌细胞增殖，从而起到生长因子的作用。我们已经表明，T3和T4发挥强大的促血管生成的作用，在人真皮微血管内皮细胞测定和鸡胚绒毛尿囊膜（CAM）和基质胶模型的血管生成。总之，这些发现表明，甲状腺激素和/或相关信号通路的拮抗剂可能是治疗癌症的有力靶点。我们实验室最近的研究表明，四碘甲状腺乙酸（tetraiodothyroacetic acid，tetrac），脱氨基的甲状腺激素类似物，能够阻止T4与1vB 3在细胞膜上的结合，对细胞增殖和血管生成产生深远的抑制作用。Tetrac抑制癌细胞增殖的发现促使我们研究它是否也可能刺激细胞对压力的反应，从而逆转耐药性的发展。我们的研究结果表明，tetrac对耐药性的影响，并涉及至少两个途径，在细胞对化疗的反应中发挥关键作用。在本探索性应用中，我们将研究Tetrac作为治疗癌症的治疗工具的潜在用途，特别是关于其在抑制化疗耐药性发展方面的可能作用。目的是确定tetrac的抗血管生成和抗增殖作用是否在质膜而不是在基因组水平上启动。为此，我们已经产生了PLGA纳米粒子共价连接到其表面上的Tetrac。初步数据显示，在与癌细胞孵育后，颗粒优先定位于质膜。假设：使用Tetrac共价连接至纳米颗粒（TclNPs）的Tetrac优先靶向质膜将提供研究这种激素拮抗剂的质膜受体介导的作用的独特方法。在拟议的研究中，我们将研究其抗血管生成和抗增殖作用。由于全身使用的甲状腺药物可能会影响多种生理途径，Tetrac选择性靶向质膜上的受体可能足以显著抑制侵袭性肿瘤的增殖，副作用较少。使用负载有化疗剂Paclitaxel的可生物降解的TclNP也可以用于在肿瘤部位控制释放该药物，因此预期导致更好的抗肿瘤活性和降低的Paclitaxel毒性。具体目标：1。将进行实验以评估TclNP在MCF 7细胞和内皮细胞中的体外功能活性。使用荧光显微镜的初步数据显示，荧光素负载的TclNP优先定位在乳腺癌MCF 7细胞的质膜上。将进行进一步的研究以确定TclNP的结合特征及其对增殖和细胞迁移的影响。还将测试TclNP对与肿瘤细胞中药物抗性的发展或逆转相关的途径的作用。将TclNP的效果与游离Tetrac在这些系统中的效果进行比较。2.在进行裸鼠的纳米颗粒靶向治疗之前，将在肿瘤生长和血管生成的鸡绒毛尿囊膜（CAM）肿瘤植入模型中的卵中进行确定TclNP的最佳制剂的先导实验。CAM模型允许在卵中预筛选生物活性，同时限制使用更敏感和昂贵的鼠物种。将在实验方法中描述的原位乳腺癌模型中测试在CAM模型中显示最佳抗肿瘤和抗血管生成活性的纳米颗粒制剂。3.雌性无胸腺小鼠将具有耐药性MCF 7人乳腺癌细胞原位植入第四乳腺中。我们将评估靶向纳米颗粒TclNPs在减少肿瘤生长和肿瘤血管生成中的有效性。我们将评估这些制剂在限制与全身性Tetrac施用相关的潜在毒性方面的功效，并确定TclNP（有或没有Paclitaxel）是否起到限制耐药性发展的作用，如我们的初步体外研究所建议的。在这些研究中评估的新型纳米颗粒系统结合了靶向和抗肿瘤活性的特性，并且由于Tetrac、Paclitazone和PLGA纳米颗粒系统都被批准用于患者，因此在可预见的未来可能会发现潜在的临床应用。 公共卫生相关性：癌细胞具有对化疗药物产生耐药性的独特能力，因此研究逆转这种现象的方法对癌症患者的治疗具有重要价值。该项目将在乳腺癌小鼠模型中使用两种药物的组合，其中一种药物会削弱癌细胞产生耐药性的能力。一种新的技术，使用纳米粒子封装测试药物，并将其引导到肿瘤将进行测试，以确定这些纳米粒子是否可以改善药物的输送和最大限度地减少相关的毒性。