DNA-linked dendrimer nanoparticle systems for cancer diagnosis and treatment

描述（由申请人提供）： 靶向的基于树枝状聚合物的纳米器件已经在体外细胞培养和体内动物研究中显示出作为癌症治疗剂的良好前景。然而，每个装置必须针对特定的靶向分子、成像剂和所需治疗剂的集合定制合成。我们提出了一个独特的解决方案，通过开发由互补寡核苷酸连接的单功能树枝状聚合物模块来解决这一限制。这允许靶向、成像和治疗性树枝状聚合物简单地通过将这些试剂的混合物加热到高于寡核苷酸双链体的退火温度而组合成多功能治疗剂。该项目将包括五个具体目标。具体目标1将涉及与聚（酰胺基）胺树枝状聚合物或树枝状分子缀合的互补寡核苷酸的设计和合成，以实现所需的结构拓扑。将使用明确定义的靶向分子（叶酸、her 2抗体和RGD肽）、药物（甲氨蝶呤、紫杉醇、顺铂和多柔比星）、成像剂（钆螯合剂和荧光染料）评估构建复杂器械的能力。具体目标2将使用包括PAGE、HPLC、CE、质谱、NMR、AFM和NSOM在内的技术来表征自组装纳米器件。具体目标3涉及测试DNA连接的纳米器件在体外的结合和内化;结合的亲合力和特异性将使用CD、差示量热法和Biacore分析来检查。将测试携带治疗剂的器械诱导细胞死亡的有效性，还将测试所有器械的固有细胞毒性。具体目标4采用动物模型来评估树状聚合物连接的治疗剂在体内治疗肿瘤的有效性。此外，将使用放射性标记材料和使用飞秒脉冲双光子激发的新型光纤探针评估治疗剂的生物分布。最后，在具体目标5下，我们将与弗雷德里克的NCI纳米颗粒表征实验室合作，使该计划中开发的材料可供其他研究人员使用。这个平台有可能彻底改变癌症治疗方法，促进“个性化医疗”。" 铺设说明：我们正在设计一种方法和工具，用于开发可以根据个体患者需求量身定制的靶向癌症药物。医生可以选择各种成分，然后将这些成分像“修补玩具”一样连接在一起，以制作个性化的药物。这种药物将选择性地只针对癌症，从而避免常规癌症化疗引起的恶心，脱发和疾病。

DESCRIPTION (provided by applicant): Targeted dendrimer-based nanodevices have shown excellent promise in both in vitro cell culture and in vivo animal studies as cancer therapeutics. However, each device must be custom synthesized for a particular set of targeting molecules, imaging agents, and desired therapeutics. We propose a unique solution to this limitation by developing single function dendrimer modules linked by complementary oligonucleotides. This allows targeting, imaging, and therapeutic dendrimers to be combined into multifunctional therapeutics simply by heating mixtures of these agents above the annealing temperature of the oligonucleotide duplex. The project will consist of five specific aims. Specific Aim 1 will involve the design and synthesis of complementary oligonucleotides conjugated to poly(amido)amine dendrimers or dendrons to achieve the desired structural topologies. The ability to construct complex devices will be assessed using well-defined targeting molecules (folic acid, her2 antibodies and RGD peptides), drugs (methotrexate, Taxol, cis-platin and doxorubicin), imaging agents (Gadolinium chelators and fluorescent dyes). Specific Aim 2 will characterize the self-assembled nanodevices using techniques including PAGE, HPLC, CE, Mass Spectroscopy, NMR, AFM, and NSOM. Specific Aim 3 involves testing the DNA-linked nanodevices for binding and internalization in vitro; the avidity and specificity of binding will be examined using CD, differential calorimetry and Biacore analyses. Devices carrying therapeutics will be tested for effectiveness at inducing cell death, and all devices will also be tested for inherent cytotoxicity. Specific Aim 4 employs animal models to assess the effectiveness of the dendrimer linked therapeutics to treat tumors in vivo. In addition, the biodistribution of the therapeutics will be assessed using radiolabeled material and a novel fiber optic probe that uses two-photon excitation with femto-second pulses. Finally, under Specific Aim 5 we will work with the NCI nanoparticle characterization lab in Frederick to make the materials developed in this program available to other investigators. This platform has the potential to revolutionize cancer therapeutics and facilitate "personalized medicine." Lay description: We are designing a method and the tools for developing targeted cancer drugs that can be tailored to the needs of an individual patient. The physician can select various components and the components are then linked together like "tinker toys" to make a personalized medicine. This medicine would selectively target only the cancer, thereby avoiding the nausea, hair loss and illness caused by regular cancer chemotherapy.