C/EBPalpha-mediated therapy for acute myeloid leukemia

C/EBPα 介导的急性髓系白血病治疗

• 批准号: 6513675
• 负责人: HANNA S RADOMSKA
• 金额: $ 9.23万
• 依托单位: BETH ISRAEL DEACONESS MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-09-15 至 2005-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6513675
• 关键词: acute myelogenous leukemia; biotechnology; bone marrow; chimeric proteins; clinical research; gene induction /repression; high throughput technology; human tissue; mixed tissue /cell culture; neoplasm /cancer therapy; protein transport; technology /technique development; transcription factor

DESCRIPTION (provided by applicant): The transcription factor C/EBPa is necessary and sufficient for neutrophil differentiation. Expression and/or function of C/EBPalpha were found to be perturbed by various mechanisms in many cases of acute myeloid leukemia (AML). Consequently, conditional expression of C/EBPa in leukemic cells re-established their neutrophilic differentiation. This proposal offers novel C/EBPalpha-mediated transcriptional therapies for AML by two approaches. In one, noninvasive delivery of functional C/EBPa protein into nuclei of diseased cells will be utilized. Cell permeable peptides will be fused in frame with C/EBPa protein and expressed in eukaryotic cells. Intracellular protein transport will be achieved through co-culture of the producer cell line with the recipient cells, or by supplying the purified cell permeable C/EBPa proteins into culture media of the recipient cells. Optimized procedure will be applied to primary leukemic bone marrow cells. The expected effects of C/EBPa protein delivery, such as induction of differentiation and apoptosis, will be monitored. In the second approach, a rapid and reproducible cell-based high throughput screen of small molecule chemicals was developed. Briefly, a stable indicator line was made, which harbors luciferase gene under the control of C/EBP-responsive element. Increase in luciferase activity will likely result from augmentation of C/EBPa expression and/or function. Active compounds will be identified and studied to determine the mechanisms of their action. Although this proposal is focused on development of new therapies for AML, successful C/EBPa targeting therapies might be also used for treatment of CML refractory to interferon` or ST1571. The applicant has considerable skills necessary to perform these experiments. Nevertheless, completion of the proposed career development program will further expand her research experience and will enable her to pursue an independent research career.

描述（由申请人提供）： 转录因子C/EBPa是中性粒细胞分化所必需和充分的。在许多急性髓性白血病（AML）病例中，发现C/EBPalpha的表达和/或功能受到各种机制的干扰。因此，C/EBPa在白血病细胞中的条件性表达重新建立了它们的嗜酸性分化。该提议通过两种方法提供了用于AML的新型C/EBPa介导的转录疗法。在一种情况下，将利用功能性C/EBPa蛋白质非侵入性递送到患病细胞的细胞核中。细胞渗透性肽将与C/EBPa蛋白框内融合并在真核细胞中表达。细胞内蛋白质转运将通过生产细胞系与受体细胞的共培养，或通过将纯化的细胞可渗透C/EBPa蛋白质供应到受体细胞的培养基中来实现。优化的程序将应用于原代白血病骨髓细胞。将监测C/EBPa蛋白递送的预期效果，例如诱导分化和凋亡。在第二种方法中，开发了一种快速且可重复的基于细胞的小分子化学品的高通量筛选。简言之，制备了稳定的指示线，其携带在C/EBP响应元件控制下的荧光素酶基因。荧光素酶活性的增加可能由C/EBPa表达和/或功能的增强引起。将对活性化合物进行鉴定和研究，以确定其作用机制。尽管该提议集中于开发AML的新疗法，但成功的C/EBPa靶向疗法也可用于治疗干扰素或ST 1571难治的CML。申请人具有进行这些实验所需的相当多的技能。尽管如此，完成拟议的职业发展计划将进一步扩大她的研究经验，并使她能够追求独立的研究生涯。