Enhancement of Tumor-Targeted Transgene Expression

增强肿瘤靶向转基因表达

• 批准号: 6736018
• 负责人: Esther H Chang
• 金额: $ 20.17万
• 依托单位: SYNERGENE THERAPEUTICS, INC.
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-09-01 至 2005-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6736018
• 关键词: atomic force microscopy; cytotoxicity; electron microscopy; enzyme linked immunosorbent assay; flow cytometry; gene delivery system; gene expression; gene therapy; laboratory mouse; liposomes; p53 gene /protein; pancreas neoplasms; reporter genes; technology /technique development; terminal nick end labeling; transfection; western blottings

DESCRIPTION (provided by applicant): In this application, the long-term goal is the development of an effective tumor-targeting lipoplex for systemic treatment of pancreatic cancer (PanCa), which has one of the worst prognosis of all human malignancies. Our previous studies indicate that cancer cells can be sensitized to radiation/chemotherapy via p53 restoration ,using a tumor-targeting liposomal gene delivery system. For the delivery of the wtp53 gene, as well as other genes, we have developed a ligand, tumor-targeting liposome complex, a nanoparticle delivery system (NDS). This resulted not only in tumor growth inhibition and/or increased survival, but also, in complete, long-term elimination of some tumors in mouse models of cancer when combined with either radiation or chemotherapy. To further achieve improvement in transfection efficiency of our NDS, recently we have designed the inclusion of a pH sensitive cationic HK peptide (histidylated oligolysine) in the complex to enhance DNA release from the endosome. One of the advantages of our NDS is that it can be broadly employed for numerous tumor types. Thus, in this application we will focus on PanCa. The major aims of this STTR I proposal are to optimize this modified complex and evaluate its transfection efficiency in vitro and its in vivo targeting ability for PanCa tumors. The assessment of this novel NDS with respect to its potential for efficient tumor-targeting delivery and antitumor efficacy can be cumbersome when based solely on biological assays, particularly in regards to product release criteria for production of therapeutic agents. Consequently, the development of methods for characterizing NDS using physical parameters would be essential. Joining forces with Dr. John Dagata at NIST, we propose to use advanced imaging analysis including atomic force microscopy, electric force microscopy, and phase imaging to characterize the NDS and correlate the results of these physical analyses with the observed biological effects of the complex. This will allow us to leverage our efforts in a pioneering attempt to bridge the gap between two distinct technologies, thus developing new engineering methodologies to characterize, design and improve therapeutic delivery systems. In addition, establishing new technologies to assess the physical characteristics of NDS will also aid in developing more precise product release criteria for these biological therapeutics.

描述（由申请人提供）：在本申请中，长期目标是开发用于胰腺癌（PanCa）全身治疗的有效肿瘤靶向脂质复合物，胰腺癌是所有人类恶性肿瘤中预后最差的肿瘤之一。我们以前的研究表明，癌细胞可以通过p53修复，使用肿瘤靶向脂质体基因递送系统对放射/化疗敏感。对于wtp 53基因以及其他基因的递送，我们已经开发了一种配体、肿瘤靶向脂质体复合物、纳米颗粒递送系统（NDS）。这不仅导致肿瘤生长抑制和/或存活率增加，而且当与放射或化学疗法组合时，在小鼠癌症模型中完全长期消除一些肿瘤。为了进一步提高我们的NDS的转染效率，最近我们设计了在复合物中包含pH敏感的阳离子HK肽（组氨酸寡聚赖氨酸）以增强DNA从内体的释放。我们的NDS的优势之一是它可以广泛用于许多肿瘤类型。因此，在本申请中，我们将重点关注PanCa。该STTR I提案的主要目的是优化这种修饰的复合物，并评估其体外转染效率及其对PanCa肿瘤的体内靶向能力。当仅基于生物测定时，特别是关于用于生产治疗剂的产品放行标准时，这种新型NDS关于其有效的肿瘤靶向递送和抗肿瘤功效的潜力的评估可能是繁琐的。因此，开发使用物理参数表征NDS的方法至关重要。与NIST的John Dagata博士合作，我们建议使用先进的成像分析，包括原子力显微镜，电力显微镜和相位成像来表征NDS，并将这些物理分析的结果与观察到的复合物的生物效应相关联。这将使我们能够利用我们的努力，开拓性地尝试弥合两种不同技术之间的差距，从而开发新的工程方法来表征，设计和改进治疗给药系统。此外，建立评估NDS物理特性的新技术也将有助于为这些生物治疗剂制定更精确的产品放行标准。