Short-Interfering RNA-Gold Nanoparticle Bioconjugates: A New Cancer Therapy

描述(申请人提供)：2006年，癌症超过心脏病成为美国的头号死因。虽然密集的研究工作导致癌症患者的5年存活率总体上有所改善，但那些被诊断为肺癌和乳腺癌的患者在生活质量和存活率方面只有名义上的改善。RNAi是一种酶介导的途径，它使用短干扰RNA寡核苷酸(SiRNAs)来特异性地降解靶序列并下调靶基因的表达。SiRNAs有望成为治疗癌症的新方法，因为下调某些在癌细胞中过度表达的基因，如Survivin，可以诱导细胞凋亡(细胞死亡)，而正常细胞不受影响。因此，这些基因是siRNA治疗的有吸引力的治疗靶点。然而，siRNA疗法的发展一直受到低效率的胞外递送和寡核苷酸摄取的阻碍。金纳米颗粒寡核苷酸生物结合物是一种无毒的反义剂，可以将寡核苷酸运送到细胞膜上，增加对核酸酶的抗性，并特异性地下调基因的表达。这项研究计划的目的是开发siRNA金纳米颗粒制剂，以沉默肺癌和乳腺癌模型中的Survivin基因，最终诱导细胞死亡。SiRNA纳米制剂的制备方法主要有固相合成法、溶液法和金硫醇化学法。这些反义剂的细胞摄取和特异性基因沉默能力将通过基因表达分析(PCR和Western Blot)对不同的模型细胞株进行定量测定。此外，除了siRNA外，还将向纳米颗粒支架中添加靶向剂，以增加癌细胞对siRNA纳米颗粒的摄取。这一治疗癌症的创新战略有可能大幅提高癌症患者的存活率和生活质量，同时利用纳米技术的根本进步来洞察疾病的功能和治疗。这项研究计划将使我能够在我的科学背景的基础上，在一个跨学科的环境中工作，探索使用纳米技术治疗和发挥疾病的功能。我希望开发和研究这个反义平台将是一次宝贵的经历，因为我作为一名大学教员开始了独立的职业生涯，他的研究致力于将纳米技术工具应用于与健康相关的问题。癌症是美国的主要死因，这种疾病的治疗是公共卫生的优先事项。这项拟议的研究使用纳米技术敲除致癌基因，这将导致治疗乳腺癌和肺癌的强大疗法。

DESCRIPTION (provided by applicant): In 2006, cancer surpassed heart disease as the leading cause of death in the United States. While intensive research efforts have led to an overall improvement in the 5-year survival rate of cancer patients, those diagnosed with lung and breast cancer have experienced only nominal improvements in quality of life and survival rate. RNAi is an enzyme-mediated pathway that uses short-interfering RNA oligonucleotides (siRNAs) to specifically degrade a target sequence and knockdown expression of the target gene. siRNAs are promising new treatments for cancer because down-regulation of certain genes, such as survivin, that are over-expressed in cancer cells can induce apoptosis (cell death), while leaving normal cells unaffected. As a result, these genes are attractive therapeutic targets for siRNA therapy. Development of siRNA therapeutics, however, has been hampered by inefficient extracellular delivery and uptake of the oligonucleotides. Gold nanoparticle oligodeoxynucleotide bioconjugates are non-toxic antisense agents that deliver oligonucleotides across the cell membrane, increase resistance to nucleases, and knockdown gene expression specifically. The objective of this research proposal is to develop siRNA gold-nanoparticle agents to silence the survivin gene in lung and breast cancer models, ultimately inducing cell-death. The siRNA nanoparticle agents will be prepared by solid phase synthesis, solution methods and gold-thiol chemistry. The cellular uptake and specific gene silencing ability of these antisense agents will be quantitatively determined for different model cell lines using gene-expression analysis (PCR and Western Blot). Additionally, targeting agents that increase the uptake of the siRNA nanoparticles by cancer cells will be added to the nanoparticle scaffold in addition to the siRNAs. This innovative strategy for the treatment of cancer has the potential to create substantive improvements in survival rates and quality of life for cancer patients, while using fundamental advances in nanotechnology to provide insight into the function and treatment of disease. This research program will allow me to build upon my scientific background by working in an interdisciplinary environment exploring the treatment and functioning of disease using nanotechnology. I expect that developing and studying this antisense platform will be a valuable experience as I start an independent career as a university faculty member whose research is devoted to applying the tools of nanotechnology to health-related issues. Cancer is the leading cause of death in the United States and treatment of this disease is a public health priority. The proposed research uses nanotechnology to knock out cancer-causing genes, which will lead to powerful therapies for treating breast and lung cancer.