MODULATION OF CELL GROWTH BY ANTISENSE AND ANTIGENE REAGENTS

通过反义和抗原试剂调节细胞生长

• 批准号: 2464446
• 负责人: L M NECKERS
• 金额: --
• 依托单位: DIVISION OF CLINICAL SCIENCES - NCI
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/2464446
• 关键词: antineoplastics; antisense nucleic acid; biological transport; bone marrow purging; cell growth regulation; gene induction /repression; human genetic material tag; human tissue; method development; neoplasm /cancer genetics; nucleic acid sequence; oligonucleotides; protooncogene; virus replication

The focus of this project is three-fold: (1) to characterize uptake and intracellular processing of unmodified and modified oligonucleotides; (2) to utilize antisense and antigene technology in several in vitro model systems to identify critical elements in cell proliferation/Viral replication; and (3) to study the efficacy of antisense and antigene reagents as in vivo modulators of gene expression. (l) We have characterized the uptake of modified oligos as an energy-dependent, endocytic process, mediated by at least one cell surface-binding protein. We have devised a novel technique to study olig uptake, intracellular localization, and association with protein and nucleic acids. This non- invasive technique will permit subcellular localization over time of an internalized oligo. (2) We have confirmed that c-myc inhibition is cytostatic for normal and malignant lymphoid cells and some Burkitt lymphoma cells can be specifically growth-arrested in vitro with a novel c-myc antisense. We have confirmed that N-myc inhibition leads to reduction in growth secondary to alteration in differentiative status of neuroectoderm-derived cell lines. (3) We have demonstrated that c-myc antisense is particularly effective against several solid tumors, including human and rat glioblastoma. In solid tumors, the c-myc antisense oligonucleotide has, in addition to its antisense effects, a sequence-specific, non-antisense mediated, effect on cellular attachment to extracellular matrix. (4) We have identified sequences capable of specifically inhibiting bcr-abl tyrosine kinase and other tyrosine kinases via direct interaction with the proteins. (5) We have been able to demonstrate significant prolongation of animal survival following injection of tumor cells treated with antisense to c-myc. At the same time such antisense treatment has no effect on normal bone marrow cells. These last findings have led to initiation of the process required to undertake a phase 1 clinical trial using antisense (c-myc-targeted) and anti-protein (bcr-abl-targeted) oligonucleotides as bone marrow purging agents.

该项目的重点有三个：(1) 描述吸收和吸收的特征 未修饰和修饰寡核苷酸的细胞内加工； (2) 在多种体外模型中利用反义和反基因技术 识别细胞增殖/病毒关键要素的系统 复制； (3) 研究反义和反基因的功效 作为体内基因表达调节剂的试剂。 (l) 我们有 将修饰寡核苷酸的吸收表征为能量依赖性， 由至少一种细胞表面结合蛋白介导的内吞过程。 我们设计了一种新技术来研究细胞内寡核苷酸的摄取 定位以及与蛋白质和核酸的关联。 这个非 侵入性技术将允许随着时间的推移进行亚细胞定位 内化寡核苷酸。 (2) 我们已经证实c-myc抑制作用是 对正常和恶性淋巴细胞以及某些 Burkitt 细胞具有细胞抑制作用 淋巴瘤细胞可以在体外用一种新的特异性生长抑制 c-myc反义。 我们已经证实 N-myc 抑制会导致 继发于分化状态改变的生长减少 神经外胚层衍生的细胞系。 (3) 我们证明了c-myc 反义对几种实体瘤特别有效， 包括人和大鼠的胶质母细胞瘤。 在实体瘤中，c-myc 反义寡核苷酸除了具有反义作用外，还具有 序列特异性、非反义介导的对细胞附着的影响 至细胞外基质。 (4) 我们已经鉴定出能够 特异性抑制 bcr-abl 酪氨酸激酶和其他酪氨酸 激酶通过与蛋白质直接相互作用。 （5）我们已经能够 证明动物存活率显着延长 注射用c-myc反义蛋白处理的肿瘤细胞。 同时 此时这种反义治疗对正常骨髓细胞没有影响。 这些最后的发现导致启动了所需的流程 使用反义（c-myc 靶向）进行 1 期临床试验，并且 抗蛋白（bcr-abl 靶向）寡核苷酸作为骨髓净化 代理。