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: 5201271
• 负责人: L M NECKERS
• 金额: --
• 依托单位: DIVISION OF CANCER TREATMENT
• 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/5201271
• 关键词: antineoplastics; antisense nucleic acid; astrocytoma; binding proteins; bioenergetics; cell adhesion; cell cycle; cell differentiation; cell growth regulation; endocytosis; extracellular matrix; gene expression; germ cells; growth inhibitors; human tissue; intracellular transport; neoplasm /cancer pharmacology; neoplastic cell; nucleotide metabolism; oligonucleotides; protein tyrosine kinase; 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 events 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.

该项目的重点有三个方面：（1）确定吸收的特点， 未修饰和修饰的寡核苷酸的细胞内加工;（2） 在几种体外模型中利用反义和反基因技术 用于识别细胞增殖/病毒中关键事件的系统 复制;（3）研究反义和反义基因的功效 作为基因表达的体内调节剂的试剂。 (l)我们有 其特征在于修饰的寡核苷酸的摄取是能量依赖性的， 内吞过程，由至少一种细胞表面结合蛋白介导。 我们设计了一种新的技术来研究olig摄取， 定位以及与蛋白质和核酸的结合。 这个非- 侵入性技术将允许随着时间推移， 内化寡核苷酸 (2)我们已经证实，c-myc抑制是 对正常和恶性淋巴细胞和某些伯基特淋巴细胞的细胞生长抑制剂 淋巴瘤细胞可以在体外特异性地生长抑制， c-myc反义核酸 我们已经证实，N-myc抑制导致 继发于分化状态改变的生长减少 神经外胚层来源的细胞系。 (3)我们已经证明c-myc 反义对几种实体瘤特别有效， 包括人类和大鼠胶质母细胞瘤。 在实体瘤中，c-myc 反义寡核苷酸除了其反义作用外， 序列特异性，非反义介导，对细胞附着的影响 细胞外基质 (4)我们已经鉴定出能够 特异性抑制bcr-abl酪氨酸激酶和其它酪氨酸激酶 通过与蛋白质的直接相互作用。 (5)我们已经能够 证明动物存活时间显著延长， 注射用c-myc反义核酸处理的肿瘤细胞。 在同一 这种反义治疗对正常骨髓细胞没有影响。