Genomics of AML: Contribution of Cytokine Signaling Pathways to Leukemogenesis

AML 基因组学：细胞因子信号通路对白血病发生的贡献

基本信息

批准号：
7465878
负责人：
MICHAEL H TOMASSON
金额：
$ 28.14万
依托单位：
WASHINGTON UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2008
资助国家：
美国
起止时间：
2008-04-01 至 2013-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7465878
关键词：
Acute Myelocytic Leukemia Affect Alleles Animal Model Appendix Biochemical Biological Biological Assay Biological Models Biological Process Bone Marrow Cell Line Cell membrane Cells Co-Immunoprecipitations Confocal Microscopy Critical Pathways Cultured Cells Cytokine Gene Cytokine Receptors Cytokine Signaling Data Depth Differentiation and Growth Disease Disease Progression FLT3 gene Family Fetal Liver Frequencies Funding Gene Mutation Genes Genetic Polymorphism Genomics Genotype Goals Growth Hematopoietic Hematopoietic stem cells Human JAK1 gene JAK2 gene JAK3 gene Janus kinase KRAS2 gene Kinase Family Gene Laboratories Mediating Mediator of activation protein Mitogen-Activated Protein Kinases Modeling Molecular Target Mus Mutation Myelogenous Myeloid Cells Myeloid Leukemia Myeloproliferative disease N-ras Genes NRAS gene NTRK3 gene Nucleotides Numbers Oncogenes PDGFRB gene Pathogenesis Pathway interactions Patients Phosphotransferases Play Population Control Protein Tyrosine Kinase Proteins Ras/Raf Receptor Protein-Tyrosine Kinases Refractory Reporting Role Sampling Signal Pathway Signal Transduction Single Nucleotide Polymorphism Somatic Mutation TYK2 Testing Transplantation Validation abl Oncogene cell transformation in vivo leukemia leukemogenesis metaplastic cell transformation mouse model mutant novel p21 N-Ras Protein protein activation receptor research study stem trafficking tumor tyrosine receptor

项目摘要

The long-term goal of Project 4 is to validate the biological significance of discovered mutations so that specific molecular targets can be used to guide the treatment of patients with acute myeloid leukemia (AMI). Mutations in genes involved in cytokine signaling (e.g. PL73, KRAS, NRAS, KIT) can be identified in nearly 50% of AML cases. We hypothesized that every AML tumor harbors at least one mutation in a cytokine signaling gene, and that these mutations cooperate to cause disease progression. Focused high-throughput exonic re-sequencing of expression-prioritized receptor tyrosine kinase (RTK), cytoplasmic tyrosine kinase (CTK), and Ras-MAPK pathway genes has been performed on 94 selected primary de novo AML patient samples. Sequence data has been analyzed and novel, non-synonymous mutations have been identified in multiple genes in the Janus kinase (JAK) family, including JAK1, JAK3, and TYK2. We propose the following Specific Aims: Specific Aim 1: We will validate the biological significance of somatic Janus kinase (JAK) family mutations by assessing the growth and differentiation of primary hematopoietic cells expressing mutant JAK1 and TYK2, and we will characterize the mechanisms by which these somatic mutations contribute to leukemic transformation. Our laboratory has validated the functional significance of several myeloid leukemia-associated oncogenes using biochemical, cell culture, and mouse model assays. We will express JAK1, TYK2, and JAK3 mutations in cell lines and primary murine bone marrow assays to determine the effect of these mutations on growth and survival of hematopoietic cells. The effects of these mutations on the subcellular localization of the mutant proteins themselves, and associated proteins will be examined using confocal microscopy, biochemical analysis, and localization-tagged mutants. Specific Aim 2: We will validate the biological significance of high-priority germline single nucleotide polymorphisms (SNPs) in JAK family genes TYK2 and JAK3 by characterizing their functions using cell culture and animal model systems. We have found known and previously unidentified nonsynonymous SNPs in our AML discovery set. To prioritize these SNPs for further study, we will collaborate with Project 5 to determine the frequency of these SNPs in control populations. We will express high-priority non-synonymous SNPs in cell lines and in murine bone marrow transduction-transplantation assays to characterize their role in transformation. We will also perform biological validation experiments with mutations and SNPs in cooperation as they are found in our patients to rigorously assess the contribution of discovered mutations to disease pathogenesis.

项目 4 的长期目标是验证已发现突变的生物学意义，以便特定的分子靶点可用于指导急性髓系白血病（AMI）患者的治疗。细胞因子信号转导相关基因（例如 PL73、KRAS、NRAS、KIT）的突变几乎可以在 50% 的 AML 病例。我们假设每个 AML 肿瘤都至少含有一种细胞因子突变信号基因，并且这些突变共同导致疾病进展。聚焦高通量表达优先受体酪氨酸激酶 (RTK)、细胞质酪氨酸激酶的外显子重测序 (CTK) 和 Ras-MAPK 通路基因已对 94 名选定的原发性新发 AML 患者进行了检测样品。对序列数据进行了分析，并在其中鉴定出了新的非同义突变 Janus 激酶 (JAK) 家族中的多个基因，包括 JAK1、JAK3 和 TYK2。我们提出以下建议具体目标：具体目标 1：我们将验证体细胞 Janus 激酶的生物学意义通过评估原代造血细胞的生长和分化来检测 (JAK) 家族突变表达突变体 JAK1 和 TYK2，我们将描述这些突变体的机制体细胞突变有助于白血病转化。我们的实验室已经验证了功能使用生化、细胞培养和小鼠研究几种髓系白血病相关癌基因的重要性模型分析。我们将在细胞系和原代小鼠骨中表达 JAK1、TYK2 和 JAK3 突变骨髓测定以确定这些突变对造血细胞生长和存活的影响。这这些突变对突变蛋白本身的亚细胞定位的影响，以及相关的将使用共聚焦显微镜、生化分析和定位标记突变体来检查蛋白质。具体目标2：我们将验证高优先级种系单核苷酸的生物学意义 JAK 家族基因 TYK2 和 JAK3 的多态性 (SNP) 通过使用以下方法表征它们的功能细胞培养和动物模型系统。我们发现了已知的和以前未识别的非同义的我们的 AML 发现集中的 SNP。为了优先考虑这些 SNP 进行进一步研究，我们将合作与项目 5 一起确定这些 SNP 在对照人群中的频率。我们将高度优先表达细胞系和小鼠骨髓转导移植试验中的非同义 SNP 描述他们在转型中的角色。我们还将进行生物验证实验在我们的患者中发现突变和单核苷酸多态性（SNP），以严格评估它们的贡献发现了疾病发病机制的突变。