GENE-SPECIFIC THERAPY OF BREAST AND PANCREATIC CANCER

乳腺癌将在1992年袭击15万美国妇女，HER2被放大 并且在约30%的晚期和早期乳腺癌中过表达， 肿瘤的 1992年胰腺癌将侵袭3万名美国男性和女性， 几乎对所有病人都是致命的 超过80%的胰腺肿瘤 第12位密码子突变的c-Ki-ras癌基因。 反义和反基因DNA 治疗提供了强有力的工具， 癌基因在恶性细胞中。 该提案的目标是具体 消除乳腺癌中HER-2过表达和突变c-Ki-ras 在胰腺癌中的表达。 三种不同的新型DNA衍生物 将在四种不同的交付模式中使用， 这将产生最大的功效。 DNA衍生物将被测试 在免疫受损小鼠中对新鲜人肿瘤异种移植物的功效。 成功的衍生物将准备用于配方研究， 小型/大型动物毒理学试验由本基金的结论 期 在这个项目中，将首先植入新鲜的人类肿瘤样本 在免疫功能低下的小鼠中。 然后对这些异种移植物进行HER- 2乳腺癌中的基因扩增和蛋白质过表达 组织. 将筛选胰腺癌异种移植物中是否存在 密码子12 c-Ki-ras突变体。 为了抑制HER-2蛋白表达， 将针对mRNA的5 '末端制备DNA（5'-dCCA ATG GAG GGG AAT），和抗寡嘌呤-寡嘧啶区的抗基因DNA， 启动子（5'-dGGTGGTGGTTGTGGT）。 抑制c-Ki-ras蛋白 表达时，将针对两种最常见的 观察到的突变体序列（5'dTAC GCC AAC AGC TCC和5'dTAC GCC ATC AGC TCC）。 对于每个序列，DNA硼烷膦酸酯，DNA二硫代磷酸酯， 和立体特异性DNA甲基膦酸酯将被合成，以及 感应和控制系统 DNA衍生物将被注射到 腹腔内、口服、溶液中微泵和静脉内 在脂质体中。

Breast cancer will attack 150,000 US women in 1992, and HER2 is amplified and overexpressed in about 30% of both advanced and early stage mammary tumors. Pancreatic cancer will attack 30,000 US men and women in 1992 and is fatal in virtually all patients. Over 80% of pancreatic tumors display c-Ki-ras oncogenes with 12th codon mutations. Antisense and antigene DNA therapeutics offer powerful tools for intervention against activated oncogenes in malignant cells. The goal of this proposal is to specifically ablate HER-2 overexpression in breast cancer, and mutant c-Ki-ras expression in pancreatic cancer. Three different novel DNA derivatives will be utilized, in four different delivery modes, to find a combination which will yield maximum efficacy. The DNA derivatives will be tested for efficacy against fresh human tumor xenografts in immunocompromised mice. Successful derivatives will be ready for formulation studies and small/large animal toxicology testing by the conclusion of this funding period. In this project, fresh human tumor samples will first be implanted in immunocompromised mice. These xenografts will then be screened for HER- 2 gene amplification and protein overexpression in the case of breast tissue. Pancreatic cancer xenografts will be screened for the presence of codon 12 c-Ki-ras mutants. To inhibit HER-2 protein expression, antisense DNAs will be prepared against the 5' end of the mRNA(5'-dCCA ATG GAG GGG AAT), and antigene DNAs against an oligopurine-oligopyrimidine region in the promoter (5'-dGGTGGTGGTTGTGGT). To inhibit c-Ki-ras protein expression, antisense DNAs will be prepared against the two most frequently observed mutant sequences (5'dTAC GCC AAC AGC TCC and 5'dTAC GCC ATC AGC TCC). For each sequence, DNA borane phosphonates, DNA phosphorodithioates, and stereospecific DNA methylphosphonates will be synthesized, as well as sense and scrambled controls. The DNA derivatives will be administered intraperitoneally, orally, and by micropump in solution, and intravenously in liposomes.