Development of gene delivery system using nanocoloid and its application for gene targeting therapy

纳米胶体基因递送系统的开发及其在基因靶向治疗中的应用

• 批准号: 18390531
• 负责人: SHINDOH Masanobu
• 金额: $ 10.76万
• 依托单位: Hokkaido University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2006
• 资助国家: 日本
• 起止时间: 2006 至 2007
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-18390531/
• 关键词: nanocoloid; cell-specific gene transfer; adrenomedullin; 遺伝子導入; EGFレセプター; 口腔扁平上皮がん; ナノテクノロジー

Trials of gene therapies have been carried out for malignant tumors and congenital gene disorders,; however, there are problems such as the low efficiency and risk for applying human being, and ideal methods of gene therapy are not yet developed.. We designed nanocoloid that enclosed expression plasmid inside and, ligand peptide outside of small particle for development of a specific gene delivery system. When We searched expression of EGFR in oral squamous cell carcinoma cell lines (9.22, HSC2, HSC3, HSC4, Ca SAS) by Western blot, and all cell lines expressed EGFR We examined the binding affinity of EGFR and synthesized EGF peptide by BireCore X. As a result, it became clear that synthesized peptide bound to EGFR. Next, we examined the induction effect of nanocoloid to oral carcinoma cell lines. The efficiency of transfection was approximately 30%, and we aimed to improve the efficiency. We next examined the effects of adrenomedullin antagonist that has the ability to inhibit angiogenesis. The vector that has the sequence for inhibitory effects on aderenomedullin reduced the Growth of tumor xenografts in nude mice.These results imply that specific gene delivery for tumor environment could be the therapeutic agents for solid cancers.

恶性肿瘤和先天性基因疾病的基因治疗试验已经展开，但存在应用于人类的效率低和风险大等问题，理想的基因治疗方法尚未开发出来。我们设计了纳米粒子，将表达载体包裹在小颗粒内，将配体多肽包裹在小颗粒外面，以开发出一种特异的基因递送系统。用免疫印迹法检测EGFR在口腔鳞癌细胞系(9.22、HSC2、HSC3、HSC4、CaSAS)中的表达，并用BireCore X检测EGFR与合成的EGF多肽的亲和力，结果表明合成肽与EGFR结合。接下来，我们检测了纳米类化合物对口腔癌细胞株的诱导作用。转染率约为30%，我们的目标是提高效率。接下来，我们研究了肾上腺髓质素拮抗剂对血管生成的抑制作用。该载体具有抑制肾上腺髓质素作用的序列，可抑制裸鼠移植瘤的生长，提示针对肿瘤环境的特异性基因传递可能成为实体瘤的治疗药物。

项目成果

Suppression of tumor growth by intra-muscular transfer of naked DNA encoding adrenomedullin antagonist

• DOI: 10.1038/sj.cgt.7700979
• 发表时间: 2007-01-01
• 期刊: CANCER GENE THERAPY
• 影响因子: 6.4
• 作者: Miseki, T.;Kawakami, H.;Kobayashi, M.
• 通讯作者: Kobayashi, M.

Expression profiles to mode of invasion and lymph-node metastasis of oral squamous cell carcinoma.

口腔鳞状细胞癌侵袭和淋巴结转移模式的表达谱。

• 发表时间: 2007
• 作者: Kondoh N ;et. al.
• 通讯作者: et. al.

ポリリン酸の細胞増殖におよぼす役割

多聚磷酸对细胞增殖的作用

• 发表时间: 2006
• 作者: 山本栄治;他
• 通讯作者: 他