CMYB DURING EARLY MYELOPOIESIS

描述(申请者描述)：申请者的职业目标 血液病理学研究与临床实践相结合是合乎逻辑的 他在致癌方面的兴趣的结果，细胞 分化和转录因子。虽然他已经有了一个富有成效的 骨骼肌细胞和B细胞转录因子的研究经验， 这一提议代表着研究对象和用途的重大变化 许多技术方法对他来说都是新奇的。 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干扰，从而导致细胞 白血病细胞周期停滞或凋亡。基于这些发现， 反义寡核苷酸治疗白血病的临床研究 细胞正在进行中。尽管c-myb是一个合理的目标，但它仍然是 对白血病的治疗不是很完美，因为它还表现为 正常的造血细胞，对它们的正常发育是必不可少的。一个 更好地了解c-myb生物学可能会发现 可能还有更好的目标。 MYB可能通过与以下物质相互作用在白血病发生中发挥作用 转录辅助因子和激活促进细胞生长的特定基因 扩散。这些辅因子和c-myb激活的基因是潜在的。 针对白血病的反义治疗的靶点。这是有可能的 其中一些靶点对某些细胞周期进程是必不可少的 白血病细胞，但在正常造血祖细胞中是必不可少的。为了测试 这就是已知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抑制 将测量正常细胞和白血病细胞。新的c-myb激活基因 将使用差异显示进行识别。这些的表达方式 基因将被用反义方法抑制，并对 将测量细胞增殖。在某些白血病中，c-myb可能是 突变的导致蛋白质相互作用或基因激活改变的。 将对原发白血病进行自发性c-myb突变筛查，这些 突变株将被分析它们对细胞增殖的影响 细胞计数或氚胸苷掺入，基因激活使用 RT-PCR或Northern印迹分析，以及辅因子相互作用 免疫共沉淀法或哺乳动物双杂交试验。

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.