Myb protein specificity in human hematopoietic cells

人类造血细胞中的 Myb 蛋白特异性

• 批准号: 7624373
• 负责人: SCOTT A. NESS
• 金额: $ 25.08万
• 依托单位: UNIVERSITY OF NEW MEXICO
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-08-01 至 2010-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7624373
• 关键词: Adenovirus Vector; Adult Acute Myeloblastic Leukemia; Affect; Alternative Splicing; Animals; Apoptosis; Base Sequence; Biological; Biological Assay; Biological Testing; Birds; Breast; C-terminal; CD34 gene; Cells; Characteristics; Clinical; Colon; DNA Binding; Data; Data Analyses; Exons; Gene Expression; Genes; Hematopoietic; Human; Investigation; Lentivirus Vector; MYB gene; Messenger RNA; Methods; Minor; Monitor; Mus; Mutation; Myeloid Cells; Myeloid Leukemia; Normal Cell; Oncogene Proteins; Outcome; Pancreas; Patients; Pattern; Proteins; Proto-Oncogene Proteins c-myb; Publishing; RNA Splicing; Reading; Research Personnel; Rodent; Sampling; Solid Neoplasm; Specificity; Structure; System; Testing; Transcript; Transcription factor genes; Variant; base; bone marrow hyperplasia; cohort; human disease; leukemia; leukemia/lymphoma; leukemogenesis; monocyte; novel; outcome forecast; progenitor; programs; research study; transcription factor; v-myb Genes

DESCRIPTION (provided by applicant): This revised application proposes to compare the activities of the wild type and variant versions of c-Myb expressed by normal cells and leukemias, and to determine their relevance to human disease. The c-myb proto-oncogene encodes a DNA binding transcription factor with latent transforming activity. Relatively minor changes to the c-Myb protein unleash its ability to cause leukemias in birds and rodents and microarray assays have shown that even minor changes to its C-terminal domains result in dramatic changes in transcriptional activity. Alternative RNA splicing generates multiple c-myb mRNAs that encode proteins with altered C-terminal domains and the alternative splicing is more abundant and more varied in leukemias than in normal cells, suggesting that variants of c-Myb may participate in leukemogenesis. In Aim 1, the structures of the most abundant variant c-myb mRNAs expressed in a representative group of normal hematopoietic cells and leukemias will be determined. To compare their activities, the normal and variant forms of c-Myb will be expressed in human monocytes and microarray assays will be used to identify telltale changes in gene expression. Aim 2 will test the biological relevance of the variant c-Myb proteins. First, alternative splicing of c-myb gene products will be studied in differentiating hematopoietic cells, then wild type and variant c-Myb proteins will be expressed in normal hematopoietic progenitors, to see if changes in differentiation or proliferation are induced. In Aim 3, quantitative assays will be used to follow the expression of normal and variant c-myb RNAs in a defined cohort of adult AML samples, and then statistical analyses will determine whether specific variants correlate with clinical parameters, prognosis or gene expression patterns in AML patient samples. The proposed experiments build on our expertise and extensive preliminary and published data and have great relevance to the study of alternative RNA splicing, the activity of Myb proteins and human leukemia.

描述（由申请人提供）：此修订后的申请建议比较正常细胞和白血病表达的野生型和变体版本的活动，并确定它们与人类疾病的相关性。 C-MYB原始癌基因编码具有潜在转化活性的DNA结合转录因子。 C-MYB蛋白释放其引起鸟类和啮齿动物和微阵列分析的性白血病的能力的相对较小的变化表明，即使对其C末端结构域的微小变化也会导致转录活性的巨大变化。替代RNA剪接会产生多个C-MYB mRNA，这些C-MYB mRNA编码具有改变C末端结构域的蛋白质，而替代剪接在白血病中比正常细胞更丰富，并且在白血病中更具变化，这表明C-MYB的变异可能参与白血病。 在AIM 1中，将确定在正常造血细胞和白血病的代表性组中表达的最丰富变体的C-MYB mRNA的结构。为了比较其活动，C-MYB的正常和变异形式将在人类单核细胞中表达，微阵列分析将用于识别基因表达的变化变化。 AIM 2将测试变体C-MYB蛋白的生物学相关性。首先，将在分化的造血细胞中研究C-MYB基因产物的替代剪接，然后将在正常造血祖细胞中表达野生型和变体C-MYB蛋白，以查看是否诱发分化或增殖的变化。在AIM 3中，定量测定将用于遵循定义的成年AML样品队列中正常和变体C-MYB RNA的表达，然后统计分析将确定特定变体在AML患者样品中是否与临床参数，预后或基因表达模式相关。 提出的实验基于我们的专业知识以及广泛的初步和发布的数据，并且与替代RNA剪接，MYB蛋白和人类白血病的活性的研究非常相关。