Sanda M G, Ayyagari S R, Jaffee E M, Epstein J I, Clift S L, Cohen L K, Dranoff G, Pardoll D M, Mulligan R C, Simons J W
Brady Urological Institute, Johns Hopkins Oncology Center, Baltimore, Maryland.
J Urol. 1994 Mar;151(3):622-8. doi: 10.1016/s0022-5347(17)35032-2.
The potential efficacy and clinical feasibility of gene therapy for prostate cancer were tested. Efficacy was tested using the Dunning rat prostate carcinoma model. Rats with anaplastic, hormone refractory prostate cancer treated with irradiated prostate cancer cells genetically engineered to secrete human granulocyte-macrophage colony-stimulating factor (GM-CSF) showed longer disease-free survival compared to either untreated control rats or rats receiving prostate cancer cell vaccine mixed with soluble human GM-CSF. A gene modified prostate cancer cell vaccine thus provided effective therapy for anaplastic, hormone refractory prostate cancer in this animal model. An evaluation of the clinical feasibility of gene therapy for human prostate cancer based on these findings was then undertaken. Prostate cancer cells from patients with stage T2 prostate cancer undergoing radical prostatectomy were first transduced with MFG-lacZ, a retroviral vector carrying the beta-galactosidase reporter gene. Efficient gene transfer was achieved in each of 16 consecutive cases (median transduction efficiency 35%, range 12 to 65%). Cotransduction with a drug-selectable gene was not required to achieve high yield of genetically modified cells. Histopathology confirmed malignant origin of these cells and immunofluorescence analysis of cytokeratin 18 expression confirmed prostatic luminal-epithelial phenotype in each case tested. Cell yields (2.5 x 10(8) cells per gram of prostate cancer) were sufficient for potential entry into clinical trials. Autologous human prostate cancer vaccine cells were then transduced with MFG-GM-CSF, and significant human GM-CSF secretion was achieved in each of 10 consecutive cases. Sequential transductions increased GM-CSF secretion in each of 3 cases tested, demonstrating that increased gene dose can be used to escalate desired gene expression in individual patients. These studies show a preclinical basis for proceeding with clinical trials of gene therapy for human prostate cancer.
对前列腺癌基因治疗的潜在疗效和临床可行性进行了测试。使用邓宁大鼠前列腺癌模型测试疗效。与未治疗的对照大鼠或接受与可溶性人粒细胞巨噬细胞集落刺激因子（GM-CSF）混合的前列腺癌细胞疫苗的大鼠相比，用经基因工程改造以分泌人粒细胞巨噬细胞集落刺激因子（GM-CSF）的照射前列腺癌细胞治疗的间变性、激素难治性前列腺癌大鼠显示出更长的无病生存期。因此，一种基因修饰的前列腺癌细胞疫苗为该动物模型中的间变性、激素难治性前列腺癌提供了有效的治疗方法。然后基于这些发现对人类前列腺癌基因治疗的临床可行性进行了评估。对接受根治性前列腺切除术的T2期前列腺癌患者的前列腺癌细胞首先用携带β-半乳糖苷酶报告基因的逆转录病毒载体MFG-lacZ进行转导。在连续16例病例中均实现了高效基因转移（中位转导效率35%，范围12%至65%）。实现高产量的基因修饰细胞不需要与药物选择基因共转导。组织病理学证实了这些细胞的恶性起源，并且对细胞角蛋白18表达的免疫荧光分析在每个测试病例中均证实了前列腺管腔上皮表型。细胞产量（每克前列腺癌2.5×10⁸个细胞）足以进入临床试验。然后用MFG-GM-CSF转导自体人类前列腺癌疫苗细胞，在连续10例病例中均实现了显著的人GM-CSF分泌。在3例测试病例中，连续转导均增加了GM-CSF分泌，表明增加基因剂量可用于提高个体患者中所需基因的表达。这些研究显示了进行人类前列腺癌基因治疗临床试验的临床前基础。
