CMYB DURING EARLY MYELOPOIESIS

早期骨髓生成期间的 CMYB

• 批准号: 6513117
• 负责人: JOHN Kim CHOI
• 金额: $ 9.43万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 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 2003-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6513117
• 关键词: CD34 molecule; DNA binding protein; acute myelogenous leukemia; antisense nucleic acid; apoptosis; cell cycle; cell differentiation; cell proliferation; cofactor; gene expression; gene mutation; growth factor receptors; hematopoietic stem cells; human tissue; immunoprecipitation; leukopoiesis; neoplastic cell; northern blottings; oligonucleotides; oncogenes; polymerase chain reaction; protein structure function; protein tyrosine kinase; transcription factor

DESCRIPTION (Applicant's Description): The applicant's career goal to combine research with clinical practice of hematopathology is a logical outcome of his c o n t inued interests in carcinogenesis, cellular differentiation, and transcription factors. While he has had a productive research experience with skeletal myocytes and B cell transcription factors, this proposal represents a significant change in research subject and uses many technical approaches that are novel to him. The expression of the hematopoietic-restricted transcription factor c-myb can be disrupted using antisense oligonucleotides and this leads to cell cycle arrest or apoptosis of leukemic cells. Based on these findings, clinical trials of antisense oligonucleotide based therapy against leukemic cells are in progress. Although c-myb is a rational target, it is still less than perfect for treatment of leukemias because it is also expressed by normal hematopoietic cells and is essential for their normal development. A better fundamental understanding of c-myb biology may identify additional and possibly better targets. MYB may play a role in leukemogenesis by physically interacting with transcription co-factors and activating specific genes that promote cellular proliferation. These co-factors and c-myb activated genes are potential targets for antisense based therapies against leukemias. It is possible that some of these targets are essential for cell cycle progression in some leukemic cells but dispensable in normal hematopoietic progenitors. To test this, the expression of known c-myb activated genes and interacting proteins will be suppressed using antisense approaches and cellular proliferation of normal and leukemic cells will be measured. Novel c-myb activated genes will be identified using differential display. The expression of these genes will be suppressed using an antisense approach and the effect on cellular proliferation will be measured. In some leukemias, c-myb may be mutated resulting in altered protein interaction or gene activation. Primary leukemias will be screened for spontaneous c-myb mutations and these mutants will be analyzed for their effects on cellular proliferation using cell counting or tritiated thymidine incorporation, gene activation using RT-PCR or northern blot analysis, and co-factor interaction using co-immunoprecipitation or mammalian two hybrid assay.

描述（申请人的描述）：申请人的职业目标 将血液病理学的研究与临床实践相结合是合乎逻辑的 他对致癌作用、细胞学等方面持续感兴趣的结果 分化和转录因子。 虽然他已经取得了丰硕成果 骨骼肌细胞和 B 细胞转录因子的研究经验， 该提案代表了研究主题和用途的重大变化 许多技术方法对他来说都是新颖的。 造血限制性转录因子c-myb的表达 可以使用反义寡核苷酸来破坏，这会导致细胞 白血病细胞的周期停滞或凋亡。 基于这些发现， 基于反义寡核苷酸的白血病治疗的临床试验 细胞正在进行中。 尽管c-myb是一个理性目标，但它仍然 对于白血病的治疗并不完美，因为它也由 正常造血细胞，对其正常发育至关重要。 一个 对 c-myb 生物学更好的基本了解可能会发现更多 以及可能更好的目标。 MYB 可能通过与以下物质的物理相互作用在白血病发生中发挥作用： 转录辅助因子并激活促进细胞生长的特定基因 增殖。 这些辅助因子和 c-myb 激活基因是潜在的 基于反义的白血病疗法的靶标。 有可能 其中一些目标对于某些细胞周期进展至关重要 白血病细胞，但在正常造血祖细胞中是可有可无的。 测试 这是已知的 c-myb 激活基因和相互作用蛋白的表达 将使用反义方法和细胞增殖来抑制 将测量正常细胞和白血病细胞。 新型 c-myb 激活基因 将使用差异显示来识别。 这些的表达 基因将使用反义方法被抑制，并且对 将测量细胞增殖。 在某些白血病中，c-myb 可能是 突变导致蛋白质相互作用或基因激活改变。 将筛查原发性白血病的自发 c-myb 突变，这些 将使用以下方法分析突变体对细胞增殖的影响 细胞计数或氚化胸苷掺入，使用基因激活 RT-PCR 或 Northern blot 分析，以及使用辅助因子相互作用 免疫共沉淀或哺乳动物两种杂交测定。