Inducible NOS as a Treatment Target in CLL

诱导型 NOS 作为 CLL 的治疗靶点

• 批准号: 6320364
• 负责人: Joe Brice Weinberg
• 金额: $ 26.56万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-06-01 至 2006-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6320364
• 关键词: apoptosis; chronic lymphocytic leukemia; clinical research; cysteine endopeptidases; enzyme activity; enzyme induction /repression; enzyme inhibitors; human subject; messenger RNA; neoplasm /cancer classification /staging; neoplasm /cancer genetics; nitrates; nitric oxide; nitric oxide synthase; nitrites; phosphomonoesterases; protein kinase; transcription factor; western blottings

DESCRIPTION: (Applicant's Abstract) Chronic lymphocytic leukemia (CLL), the most common form of leukemia in North America and Europe, accounts for more than 30 percent of all cases. CLL is characterized by the accumulation of non-dividing CD5+ B cells in GO of the cell cycle. This is currently an incurable malignancy; there is a great need for new insights into disease mechanisms. Nitric oxide (NO) is produced in vivo from L-arginine by the enzyme NO synthase (NOS). NO has multiple actions that play important roles in physiology and pathology. Recent work has demonstrated that NO inhibits apoptosis. We have discovered that human CLL cells spontaneously express high levels of inducible nitric oxide synthase (NOS2) mRNA and protein, and have high NOS enzyme activity. When the CLL cells are cultured with NOS inhibitors, there is high level cell killing associated with apoptosis. The specific aims of the proposal are to: (1) Determine if in vivo NO production is increased in patients with CLL, and whether in vivo NO production correlates with NOS2 expression in CLL cells, stage of disease, and subsets of disease. We will measure serum and urine levels of the stable NO catabolites nitrite and nitrate in patients with CLL, and correlate levels with CLL cell NOS activity & NOS2 antigen content and clinical stage of disease. (2) Determine the sensitivity of patients' CLL cells to a variety of NOS inhibitors and NO quenchers. We will analyze CLL cells in vitro for viability and apoptosis after treatment with selected NOS (NOS2 specific and nonspecific) inhibitors and NO quenchers. We will also investigate the effects of combining NOS inhibitors & NO quenchers with conventional chemotherapeutic agents. (3) Determine the mechanism by which NOS2 expression is increased in CLL cells. We will determine whether the increases in NOS2 mRNA expression are due to alterations in mRNA transcription or stabilization, and whether there is constitutive activation of transcription factors known to alter NOS2 mRNA transcription. NOS2 and NO represent new molecular targets for treatment in CLL, Accomplishment of the proposed aims may provide a deeper understanding of the mechanisms of disease in CLL. Furthermore, since NOS inhibitors and NO quenchers/scavengers are now being used safely in early trials in normal volunteers and in patients with other diseases, our work may lead to rapid initiation of trials with these agents in CLL patients.

描述：（申请人摘要）慢性淋巴细胞白血病（CLL）， 北美和欧洲最常见的白血病形式，占更多 占所有病例的 30% 以上。 CLL的特点是积累 细胞周期 GO 中的非分裂 CD5+ B 细胞。目前这是一个 无法治愈的恶性肿瘤；非常需要对疾病有新的见解 机制。一氧化氮 (NO) 在体内由 L-精氨酸通过酶产生 一氧化氮合酶（NOS）。 NO 具有多种作用，在 生理学和病理学。最近的研究表明，NO 抑制 细胞凋亡。我们发现人类 CLL 细胞自发地高表达 诱导型一氧化氮合酶 (NOS2) mRNA 和蛋白质水平，并且具有 NOS酶活性高。 当CLL细胞与NOS抑制剂一起培养时，有高水平的细胞 与细胞凋亡相关的杀伤。该提案的具体目标是： (1) 确定 CLL 患者体内 NO 产生是否增加，以及 体内NO产生是否与CLL细胞中NOS2表达相关， 疾病的阶段和疾病的子集。我们将测量血清和尿液 CLL 患者稳定的 NO 分解代谢物亚硝酸盐和硝酸盐的水平， 并将水平与 CLL 细胞 NOS 活性和 NOS2 抗原含量相关联 疾病的临床阶段。 (2)测定患者CLL细胞的敏感性 多种 NOS 抑制剂和 NO 猝灭剂。我们将分析 CLL 细胞 用选定的 NOS（NOS2 特异性和非特异性）抑制剂和 NO 猝灭剂。我们也会调查 NOS抑制剂和NO猝灭剂与常规组合的效果 化疗剂。 (3)确定NOS2表达的机制 CLL 细胞中增加。我们将确定 NOS2 mRNA 是否增加 表达是由于 mRNA 转录或稳定性的改变造成的，并且 是否存在已知的转录因子的组成型激活 改变 NOS2 mRNA 转录。 NOS2 和 NO 代表了 CLL 治疗的新分子靶点， 实现所提出的目标可以使人们更深入地了解 CLL 的疾病机制。此外，由于 NOS 抑制剂和 NO 猝灭剂/清除剂现已在正常情况下的早期试验中安全使用 志愿者和患有其他疾病的患者，我们的工作可能会导致快速 开始在 CLL 患者中使用这些药物进行试验。