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）：最近提供的证据表明，甲状腺激素也可能在癌变中起作用，并且该领域的开拓性研究表明，甲状腺激素（例如3,5,3'-triioodo-l-硫代硫代硫代氨酸（T3）（T3）（T3）和L-甲状腺素（T4）可以刺激癌症的生长量。我们已经表明，T3和T4在人真皮微血管内皮细胞分析以及血管生成的鸡绒毛膜膜（CAM）和基质模型中都具有有效的促血管生成作用。综上所述，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代表了治疗癌症的引人注目的靶标。我们实验室的最新研究表明，四甲状腺乙酸（​​Tetrac）是一种脱氨基甲状腺激素类似物，能够防止T4与1VB3在细胞膜上的结合，从而对细胞增殖和血管生成产生了深远的抑制作用。 Tetrac抑制癌细胞增殖的发现促使我们研究了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。我们的结果表明，Tetrac对耐药性的影响至少牵涉到至少两种在细胞对化学疗法反应中起关键作用的途径。在此探索性应用中，我们将研究Tetrac作为治疗癌症的治疗工具的潜在用途，尤其是在抑制化学疗法抗性的发展中的作用方面。目的是确定tetrac的抗血管生成和抗增殖作用是在质膜上而不是在基因组水平上启动的。为此，我们共同生成了PLGA纳米颗粒 - 与其表面上的Tetrac相连。初步数据表明，在与癌细胞孵育后，颗粒优先定位于质膜。假设：使用与纳米颗粒的共连接（TCLNPS）将TETRAC靶向质膜（TCLNPS）将为研究这种激素拮抗剂的质膜受体介导的作用提供独特的方法。在拟议的研究的背景下，我们将研究其抗血管生成和抗增殖作用。由于系统使用的甲状腺剂可能会对多种生理途径产生潜在影响，因此在质膜处将Tetrac靶向Tetrac对受体的选择性靶向可能足以显着抑制副作用较少的攻击性肿瘤的增殖。用化学治疗剂紫杉醇加载的可生物降解的TCLNP也可以用于在肿瘤部位受控释放该药物，因此预计会导致抗蛋白酶毒性降低的抗肿瘤活性更好。具体目的：1。将进行实验，以评估MCF7细胞和内皮细胞中TCLNPS在体外的功能活性。使用荧光显微镜的初步数据表明，荧光素加载的TCLNP优先在乳腺癌MCF7细胞的质膜上定位。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。 TCLNP的作用还将在与肿瘤细胞中耐药性发育或逆转相关的途径上进行测试。在这些系统中，将将TCLNP的效果与游离Tetrac的效果进行比较。 2。确定TCLNP的最佳配方的试验实验将在OVA中在鸡肉绒毛膜膜膜（CAM）肿瘤植入物模型的肿瘤生长模型和血管生成模型中进行，然后在纳米颗粒靶向裸鼠的治疗进行之前。 CAM模型允许在OVA预筛查中获得生物活性，同时限制了使用更有意义和昂贵的鼠类。在CAM模型中显示出最佳抗肿瘤和抗血管生成活性的纳米颗粒制剂将在实验方法中描述的原位乳腺癌模型中进行测试。 3。雌性小鼠将抗药性MCF7人乳腺癌细胞原始植入第四个乳腺。我们将评估靶向纳米颗粒TCLNP的有效性，从而减少肿瘤生长和肿瘤血管生成。我们将评估这些制剂在限制与全身性tetrac给药相关的潜在毒性方面的疗效，并确定tclnps是否有或没有紫杉醇的作用是限制限制药物耐药性的发展，如我们的初步体外研究所表明的那样。在这些研究中要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，Tetrac，Paclitaxel和Plga纳米颗粒系统都被批准用于患者，因此可以在可预见的将来找到潜在的临床应用。 公共卫生相关性：癌细胞具有发展对化学治疗药物的耐药性的独特能力，因此，对扭转这种现象的方法的研究将在癌症患者的治疗中具有重要价值。该项目将使用两种药物的组合，其中一种会损害癌细胞在乳腺癌的小鼠模型中损害癌细胞发展耐药性的能力。一种新的技术，即将纳米颗粒封装测试药物，并将其引导到肿瘤中，以确定这些纳米颗粒是否可以改善药物的递送并最大程度地减少相关的毒性。