2011-08-012024-05-13https://scholars.lib.ntu.edu.tw/handle/123456789/646510摘要：基因治療可以將基因轉殖至體內特定的位置，達到特定區域內高度表現某種治療用蛋白的效果，對於腫瘤治療頗具前瞻性。之前之基因治療策略大多採用病毒載體作為基因轉殖的工具，乃因其效率較高。基因治療之成效已見曙光，但最近也因病毒載體引發之插入性突變或過度性免疫反應的問題，造成某些病人產生血癌甚或死亡，而引起大眾對基因治療之可靠性產生疑慮。相對地，非病毒基因轉殖方法就比較安全，雖然它的轉殖效率比病毒載體低。近年來，已經有很多非病毒性的基因轉殖方式被提出，超音波就是其中一個新的方式。配合顯影劑的使用，超音波的轉殖效率可以大大的被提高。本計畫目的有三；(一)、利用超音波技術轉殖抗血管新生基因，包括endostatin (ED)及calreticulin(CRT)兩種於動物肌肉組織細胞中，藉由肌肉細胞表現出抗血管新生蛋白質，並由血流將這些蛋白質傳送到遠端的目標區域，如原位腫瘤處，以抑制其血管新生。我們將以三種不同原位腫瘤之動物模式，包括肝腫瘤，肺腫瘤及腦瘤，來測試肌肉細胞在超音波轉殖抗血管生成基因後所產生之產物，是否能抑制這些原位腫瘤的生長。(二)、為增加治療效果，我們將把上述超音波基因轉殖技術與免疫治療或化療法合併，以測試是否可達到加成性的抗腫瘤效果；(三)為延長治療時效，我們將探討多種抗血管因子合併使用時，可否減少腫瘤產生抗血管新生治療之抗性；為進一步延長治療時效，我們也將測試ED及CRT 製備成Fc 之融合蛋白形式，或將ED 或CRT 與非病毒性載體結合，如liposome或 PEI(ExGen)，以期延長其在體內或肌肉細胞內的半衰期，達到減少治療之次數，和增加抗腫瘤的療效。<br> Abstract: Gene therapy is a newly developed biomedical technique that can target gene expression atspecific site, leading to a locally high expression of the therapeutic proteins, thusproviding great potentials for cancer treatment. In the past, viral vectors are predominantlyused for gene therapy due to their high transduction efficiency. However, the issuesconcerning about the safety and the immunogenecity of viral vectors have recently beenraised because of the leukemia and the fatal cases caused by the use of viral vectors. Onthe other hand, nonviral gene transfer is a safer and easier gene delivery method comparedto viral vectors, though the transduction efficiency is relatively low. In recent years, manynon-viral transfection systms have been developed for efficient gene delivery. Thepotential of therapeutic ultrasound (TUS) in combination with contrast agent to enhancegene transduction efficiency has recently received great attention. In this study, we plan todeliver antiangiogenic genes, i.e., endostatin (ED) or calreticulin (CRT), to the musclevia TUS exposure to treat distant orthotopic tumors (i.e., tumors growing in theirauthentic tissues). Three specific aims are proposed herein: (1).To evaluate the antitumoreffects of intramuscular, TUS-mediated transfer of antiangiogenic genes on distantorthotopic tumors, e.g., liver tumor, lung tumor, or brain tumor; (2). To evaluate theenhancement of antitumor effects by combining TUS-mediated antiangiogenic genetherapy with immunotherapy or chemotherapy; (3). (3). To prolong tumor therapy ofTUS-mediated antiangiogenic gene transfection, antiangiogenic gene will be use to couplewith multiple antiangiogenic factors, Fc domain of IgG or non-viral vecor such asliposome or PEI(ExGen).超音波基因轉殖聚乙烯亞胺