Controlled Release of Macromolecules

描述(由申请人提供)：这是对蛋白质和核酸等高分子量制剂配方研究的续批。在过去的29年里，这笔特别的赠款一直是我们药物输送研究的核心。我们相信，我们NIH资助的研究在这一领域的影响是巨大的，我们的工作帮助提供了一些大分子控制释放和局部药物释放的临床开发所必需的基本概念。这种送货系统的例子有Lupron Depot、Garia del和Nutroin Depot。在撰写我们最后一份竞争性续签拨款提案时，我们认为多肽和蛋白质释放的许多基本原理已经建立，最大的影响可能来自于延长我们的研究核酸输送。我们在这一领域的研究导致了更大的机械性理解，并产生了35篇同行评议的论文，包括发表在《科学》和《自然》等期刊上的论文。基因治疗在临床上取得成功的主要障碍是缺乏安全有效的DNA输送方法。经过修饰的病毒虽然能有效地将DNA转移到细胞中，但也存在严重的毒性和生产问题。相比之下，非病毒系统提供了许多重要的潜在优势，包括生产容易、稳定性、低毒性和减少载体大小限制。从我们的研究中我们发现，最佳的细胞内给药到体内不同的组织需要不同的材料。因此，我们建议开发新材料的组合库，以产生临床上有用的、用于基因治疗的非病毒方法。肺癌将被用作模式疾病。我们建议探索两种互补的方法：1)开发肺肿瘤细胞特异性DNA递送系统用于抗肿瘤治疗；2)通过靶向和激活天然免疫细胞来开发抗肺癌DNA疫苗。我们的具体目标是：1.开发下一代可生物降解聚合物，用作肺癌和DNA疫苗的高效无毒载体。2.肺肿瘤细胞特异性DNA递送系统及树突状细胞靶向研究3.检测以PEI和PBAE为载体的生物可降解载体及其静电配基包裹的三元复合载体在正常小鼠和肺癌小鼠模型中的体内基因传递效率。4.检测基于PEI和PBAE的载体作为DNA疫苗的潜力。 公共卫生相关性：基因治疗在临床上取得成功的主要障碍是缺乏安全和有效的DNA输送方法。因此，我们建议开发新材料的组合库，以产生临床上有用的、用于基因治疗的非病毒方法。

DESCRIPTION (provided by applicant): This is a grant renewal for studies on formulations of high molecular weight agents, such as proteins and nucleic acids. For the past 29 years, this particular grant has been central for our drug delivery research. We believe the impact of our NIH funded research in this area has been significant and that our work has helped provide some of the fundamental concepts necessary for the clinical development of controlled release of macromolecules and for localized drug delivery. Examples of such delivery systems are Lupron Depot, Gliadel and Nutropin Depot. In writing our last competing renewal grant proposal we felt that much of the fundamentals of peptide and protein release had been established and that the greatest impact could come from extending our research nucleic acid delivery. Our studies in this area have led to a greater mechanistic understanding, and have resulted in 35 peer-reviewed papers, including in such journals such as Science and Nature. The major barrier to the success of gene therapy in the clinic is the lack of safe and efficient DNA delivery methods. Modified viruses, while effective at transferring DNA to cells, suffer from serious toxicity and production problems. In contrast, non-viral systems offer a number of significant potential advantages, including ease of production, stability, low toxicity, and reduced vector size limitations. From our studies we have discovered that optimal intracellular delivery to different tissues in the body requires different materials. Therefore, we propose to develop combinatorial libraries of new materials with the specific goal of generating clinically useful, non-viral methods for gene therapy. Lung cancer will be used as a model disease. We propose to explore two complementary approaches: 1) developing lung-tumor cell specific DNA delivery systems for the anti-tumor therapy and 2) developing anti-lung cancer DNA vaccines by targeting and activating native immune cells. Our specific aims are: 1. To develop next-generation biodegradable polymers for use as efficient and non-toxic vectors for lung cancer and DNA vaccines. 2. Development of cell specific DNA delivery systems for lung tumor and dendritic cell targeting. 3. To test the in vivo gene delivery efficiency of biodegradable vectors composed of PEI- and PBAE- based polymer and that of their electrostatic ligand coated ternary polyplexes in normal and lung cancer mouse models. 4. To examine potential of PEI- and PBAE- based vectors for their use as DNA vaccines. Public Health Relevance: The major barrier to the success of gene therapy in the clinic is the lack of safe and efficient DNA delivery methods. Therefore, we propose to develop combinatorial libraries of new materials with the specific goal of generating clinically useful, non-viral methods for gene therapy.