MYOBLAST TRANSFER FOR DELIVERY OF ANTINEOPLASTIC AGENTS

用于递送抗肿瘤药物的成肌细胞转移

• 批准号: 6300447
• 负责人: LAURENCE H KEDES
• 金额: $ 18.41万
• 依托单位: UNIVERSITY OF SOUTHERN CALIFORNIA
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-06-01 至 2000-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6300447
• 关键词: analog; antineoplastics; breast neoplasms; cell transplantation; complementary DNA; cytokine; gene therapy; genetic enhancer element; genetic promoter element; laboratory mouse; metastasis; muscle transplantation; myoblasts; neoplasm /cancer therapy; nonhuman therapy evaluation; recombinant proteins; somatostatin; tissue /cell culture; tissue inhibitor of metalloproteinases; transfection; transfection /expression vector; xenotransplantation

This pre-clinical project will explore the unique biological attributes of muscle cells for their potential to act as gene therapy delivery vehicles for the continuous production of anti-neoplastic recombinant polypeptides in the treatment of malignancies. The overall hypothesis to be tested is that muscle cells can be engineered to secrete high levels of active polypeptides that can effect tumor growth and metastasis. Two forms of delivery will be evaluated: systemic delivery from cells implanted in skeletal muscle and local delivery from cells implanted at the sites of tumors. The effectiveness of myogenic cells will be compared with that of fibroblasts. Stable myogenic cell lines or primary myoblasts producing and secreting antineoplastic peptides in vitro will be created by transduction of expression vectors for several polypeptides including tissue inhibitors of metalloproteases (TIMPs) and somatostatin analogs (SSA) and, in collaboration with Project 4, cytokine genes. To insure muscle specific and high level expression, a series of combinations of muscle-specific enhancer and promoter elements will be evaluated in an efficient cell culture test system. The best performing promoter will he used for transduction of primary myoblasts and compared with expression from two standard promoters (CMV and beta-actin) and, in collaboration with Project 6, with a hypoxia responsive GRP78 promoter. Myogenic conversion of primary non-muscle cells, by the transduction or transfection of cDNAs expressing myogenic determination factors, may provide a readily available, expandable and renewable primary cell substrate for myoblast transfer gene therapy. The capacity of such cells to serve as constitutive secretors of recombinant polypeptides will also be tested. These experiments will attempt to determine the therapeutic effects in vivo of TIMPs and cytokine genes delivered locally by genetically engineered myoblasts in pre-tumor implantation models and established tumor growth models. The planned experiments will also determine the in vivo therapeutic efficacy against human breast cancer of systemically delivered SSA by transduced myoblasts in a pre-tumor implantation model, in an established tumor growth model and in a model of metastatic disease following the surgical removal of the primary tumor.

这个临床前项目将探索独特的生物学特性 肌肉细胞可能充当基因治疗递送车辆的潜力 为了连续产生抗塑性重组多肽 在治疗恶性肿瘤中。要测试的总体假设是 可以设计肌肉细胞以分泌高水平的活跃 可以影响肿瘤生长和转移的多肽。两种形式 将评估交付：从植入的细胞中进行全身传递 骨骼肌和局部从植入的细胞中分娩 肿瘤。将肌原细胞的有效性与 成纤维细胞。稳定的肌原细胞系或产生原发性肌细胞和原发性肌细胞和 在体外分泌的抗肿瘤肽将通过转导创建 包括组织抑制剂在内的几种多肽的表达矢量 金属蛋白酶（TIMP）和生长抑素类似物（SSA）以及在 与Project 4 Cytokine基因合作。确保特定肌肉 和高水平的表达，一系列肌肉特异性的组合 增强子和启动子元素将在有效的细胞中评估 文化测试系统。他使用的表现最好的发起人 原代成肌细胞的转导，并与两个表达进行比较 标准启动子（CMV和β-肌动蛋白），并与项目合作 6，缺氧反应性GRP78启动子。肌源性转化 原代非肌肉细胞，通过转染CDNA的转染或转染 表达肌源性的确定因素，可能会轻松提供 可用于成肌细胞的可用，可扩展和可再生的原始细胞基板 转移基因治疗。此类细胞充当本构的能力 重组多肽的捐助者也将进行测试。这些 实验将尝试确定体内的治疗作用 TIMPS和细胞因子基因在本地通过基因工程进行的 肿瘤前植入模型和确定肿瘤生长的成肌细胞 型号。计划的实验还将确定体内 针对人类乳腺癌的治疗功效 SSA通过在肿瘤前植入模型中转导的成肌细胞，在 建立的肿瘤生长模型和转移性疾病模型 手术切除原发性肿瘤后。