MEASURING c-Abl ACTIVITY IN CML USING A HIGH-AFFINITY NANOSENSOR

使用高亲和力纳米传感器测量 CML 中的 c-Abl 活性

• 批准号: 7085849
• 负责人: Robert Gellibolian
• 金额: $ 11.23万
• 依托单位: N-ABL THERAPEUTICS, INC.
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-09-14 至 2008-03-13
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7085849
• 关键词: chronic myelogenous leukemia; diagnosis; neoplasm /cancer; peptides; phosphates; phosphorylation; protein kinase; protein sequence

DESCRIPTION (provided by applicant): The ability to monitor and measure protein kinase activity in tumorigenesis and cancer can be indicative of and critical to the transformation process and therefore represents an attractive diagnostic strategy, with a multi-billion dollar market opportunity. In this proposal, we aim to develop a novel, high affinity fluorescent nanosensor and homogeneous assay system for monitoring the phosphorylation of a bona fide peptide substrate by c-Abl, an important kinase involved in the etiology of chronic myelogenous leukemia (CML), using FRET. The technology deployed during this phase of the proposal will directly translate into the development of a sensitive platform for the diagnosis of CML. In specific aim 1, we will synthesize the high-affinity nanosensor. In specific aim 2, we will develop a sensitive 1-step assay using this reagent. Specific aim 3 will elaborate on this and test the effectiveness of the nanosensor in measuring c-Abl activity in CML-positive cell lysates. Given the fact that ~400 disorders such as cancer have been associated with protein kinases, the development of a family of sensitive nanosensors such as the 1 proposed in this grant application will facilitate the diagnosis of these diseases sensitively and selectively and therefore becomes of paramount importance and will find immense utility in all the various facets of our healthcare, from drug-discovery to patient health and point-of-care diagnostics.

描述（由申请人提供）：监测和测量肿瘤发生和癌症中蛋白激酶活性的能力可以指示转化过程并对其至关重要，因此代表了具有数十亿美元市场机会的有吸引力的诊断策略。在这项提案中，我们的目标是开发一种新的，高亲和力的荧光纳米传感器和均匀的检测系统，用于监测磷酸化的一个真正的肽底物的c-Abl，一个重要的激酶参与慢性粒细胞白血病（CML）的病因，使用FRET。在该提案的这一阶段部署的技术将直接转化为诊断CML的敏感平台的开发。具体目标一是合成高亲和性的纳米传感器。在具体目标2中，我们将使用该试剂开发一种灵敏的一步测定法。具体目标3将对此进行详细说明，并测试纳米传感器在测量CML阳性细胞裂解物中c-Abl活性方面的有效性。鉴于癌症等约400种疾病与蛋白激酶有关，开发一系列敏感的纳米传感器，例如本资助申请中提出的1种传感器，将有助于灵敏地、选择性地诊断这些疾病，因此变得至关重要，并将在我们医疗保健的各个方面发现巨大的用途，从药物发现到患者健康和护理点诊断。