"BCR/ABL-PI-3k-ROS pathway induce genomic instability ...."

“BCR/ABL-PI-3k-ROS 通路导致基因组不稳定......”

• 批准号: 8271268
• 负责人: TOMASZ SKORSKI
• 金额: $ 30.8万
• 依托单位: TEMPLE UNIV OF THE COMMONWEALTH
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-07-01 至 2015-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8271268
• 关键词: 1-Phosphatidylinositol 3-Kinase; Acute Lymphocytic Leukemia; Amino Acid Substitution; Apoptosis; Appearance; Benign; Biological; Biological Assay; Blast Phase; Cell Proliferation; Cells; Chromosomal translocation; Chromosome abnormality; Chronic Myeloid Leukemia; Chronic Phase; Chronic-Phase Myeloid Leukemia; Complementary DNA; Complex; DNA Repair; Development; Disease; Disease Progression; Dominant-Negative Mutation; Drug resistance; Extracellular Matrix; Figs - dietary; Generations; Genetic; Genomic Instability; Growth Factor; Hematopoietic; Hematopoietic stem cells; Imatinib; Imatinib mesylate; In Vitro; Knockout Mice; Laboratories; Malignant - descriptor; Mediating; Mitochondria; Molecular; Mutation; Myelogenous; NADPH Oxidase; Oncogenes; Oncogenic; Pathway interactions; Patients; Perifosine; Phosphatidylinositols; Phosphotransferases; Play; Point Mutation; Population; Prevention; Production; Protein Tyrosine Kinase; Protein-Serine-Threonine Kinases; Proteins; Reactive Oxygen Species; Reporting; Resistance; Resistance development; Respiratory Chain; Role; Signal Pathway; Small Interfering RNA; Source; Stem cells; Therapeutic Effect; Tyrosine Kinase Inhibitor; Work; bcr-abl Fusion Proteins; cohort; granulocyte; improved; inhibitor/antagonist; leukemia; leukemic stem cell; leukemogenesis; macrophage; mouse model; mutant; novel strategies; oxidative DNA damage; prevent; progenitor; rac GTP-Binding Proteins; research study; small molecule; stem

Chromosomal translocation t(9;22) is responsible for appearance of an oncogene encoding BCR/ABL fusion tyrosine kinase, which induce chronic myelogenous leukemia (CML) and a cohort of acute lymphocytic leukemia (ALL). CML usually starts as a relatively benign chronic phase (CML-CP), which progresses to an aggressive disease - blast crisis (CML-BC). Malignant transformation of the disease is associated with accumulation of additional genetic errors. Imatinib mesylate (IM), a small molecule inhibitor of BCR/ABL kinase, revolutionized the treatment of CML-CP. Unfortunately patients may develop resistance to the drug caused by point mutations encoding amino acid substitutions in the BCR/ABL kinase domain. In conclusion, CML cells display genomic instability leading to resistance to IM and malignant progression of the disease. This proposal is focused on determination the mechanisms responsible for these phenomena and subsequently on prevention/inhibition of the development of IM resistance and CML-BC. BCR/ABL kinase stimulates numerous signaling pathways to induce and maintain transformation of hematopoietic cells. We, and others found that phosphatidylinositol-3 kinase (PI-3k) play an essential role in growth factor independent proliferation and protection from apoptosis in CML. Here we propose to study the role of PI-3k and its downstream effectors Akt and Rac in genomic instability in CML stem and progenitor cell populations. Using genetic approach (dominant-negative mutants, siRNA, antisense cDNA, knockout mice) and small molecule inhibitors (for example perifosine, NSC23766) we will determine how PI-3k and its downstream effectors generate the reactive oxygen species (ROS), which in turn may cause oxidative DNA damage and facilitate genomic instability. Long-term in vitro culture and mouse models of CML will be applied here. IM resistance will be detected in clonogenic assays and by sequencing BCR/ABL kinase domain, and chromosomal aberrations will be detected by SNPs and SKY. If successfully accomplished, these experiments will determine if PI-3k pathway inhibitors should be used to improve therapeutic effect of IM and prevent/delay CML progression toward lethal blast crisis.

染色体易位t（9; 22）负责癌细胞编码BCR/ABL融合的外观 酪氨酸激酶，诱导慢性粒细胞性白血病（CML）和一群急性淋巴细胞 白血病（全部）。 CML通常以相对良性的慢性相（CML-CP）开始 侵略性疾病 - 爆炸危机（CML -BC）。疾病的恶性转化与 额外遗传错误的积累。 伊马替尼梅赛酸酯（IM）是BCR/ABL激酶的小分子抑制剂，彻底改变了治疗的处理 CML-CP。不幸的是，患者可能会产生对点突变引起的药物的抗性 BCR/ABL激酶结构域中的氨基酸取代。 总之，CML细胞显示出基因组不稳定性，导致对IM和恶性肿瘤的抗性 疾病的进展。该提案的重点是确定负责这些的机制 现象以及随后的预防/抑制IM耐药性和CML-BC的发展。 BCR/ABL激酶刺激了许多信号通路，以诱导和维持的转化 造血细胞。我们和其他人发现，磷脂酰肌醇3激酶（PI-3K）在 生长因子独立的增殖和CML中凋亡的保护。 在这里，我们建议研究PI-3K及其下游效应子AKT和RAC在基因组中的作用 CML茎和祖细胞群体中的不稳定性。使用遗传方法（主要阴性突变体， siRNA，反义cDNA，基因敲除小鼠）和小分子抑制剂（例如perifosine，NSC23766）We 将确定PI-3K及其下游效应子如何产生活性氧（ROS），该物种在 转弯可能会导致氧化DNA损伤并促进基因组不稳定性。长期体外培养和小鼠 CML的模型将在此处应用。在克隆性测定中将检测到IM电阻，并通过测序 BCR/ABL激酶结构域和SNP和Sky将检测到染色体畸变。 如果成功完成，这些实验将确定PI-3K途径抑制剂是否应为 用于改善IM的治疗作用，并防止/延迟CML向致命的爆炸危机发展。