Multiplexed Kinase Biosensor Technology to Detect Leukemia Signaling with Mass Sp

多重激酶生物传感器技术通过 Mass Sp 检测白血病信号传导

• 批准号: 8930087
• 负责人: Laurie L. Parker
• 金额: $ 32.24万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-09-19 至 2017-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8930087
• 关键词: ABL1 gene; Address; Adoption; Advanced Development; Amino Acids; Antibodies; Antineoplastic Agents; Award; Bcr-Abl tyrosine kinase; Biology; Biosensor; Cancer Center; Cancer Patient; Cell Line; Cells; Chromosomes, Human, Pair 9; Chronic Myeloid Leukemia; Clinic; Clinical; Clinical Trials; Code; Collaborations; Collection; Cost Control; Detection; Development; Disease remission; Dose; Drug resistance; Exploratory/Developmental Grant for Diagnostic Cancer Imaging; Face; Family; Funding; Future; Generic Drugs; Gleevec; Gold; Health; Hematologic Neoplasms; Imatinib; Indiana; Individual; Isotopes; JAK2 gene; Label; Laboratories; Learning; Legal patent; Life; Malignant Neoplasms; Marketing; Mass Spectrum Analysis; Measures; Methodology; Methods; Monitor; Outcome; Patient Care; Patients; Peptides; Pharmaceutical Preparations; Pharmacodynamics; Philadelphia Chromosome; Phosphopeptides; Phosphorylation; Phosphotransferases; Physicians; Primary Cell Cultures; Protein Kinase Inhibitors; Proteins; Protocols documentation; Publishing; Reagent; Relapse; Relative (related person); Reproducibility; Resistance; Resistance development; Sampling; Signal Transduction; Staging; Standardization; Techniques; Technology; Therapeutic; Time; Tissue Banking; Tissue Banks; Transcript; Treatment Failure; Universities; Validation; Western Blotting; Work; anticancer research; base; bcr-abl Fusion Proteins; cancer therapy; chemotherapy; cost; drug development; drug discovery; drug market; improved; inhibitor/antagonist; instrumentation; kinase inhibitor; leukemia; leukemia treatment; multiple reaction monitoring; next generation; oncology; pre-clinical; protein kinase inhibitor; response; src-Family Kinases; success; technology development; technology validation

DESCRIPTION (provided by applicant): Kinase inhibitor drugs have changed the face of chemotherapy and are projected to remain a major focus of leukemia treatment. However, their long-term efficacy is variable and can be attributed to differences in their activity on their targts from patient to patient that can be observed as early as one week after beginning treatment. We plan to validate a technology that enables quantification of efficacy at the very beginning of treatment, giving physicians and patients the opportunity to monitor an individual's response and likelihood of success immediately, rather than relying on the current watch-and-wait approach. We have established proof-of-concept for analyzing Bcr-Abl kinase activity and inhibition in chronic myelogenous leukemia (CML) with the support of an IMAT R21 award to the Parker laboratory. An IMAT award to the Turk laboratory at Yale University also supported work leading to a collaborative effort on proof-of-concept for developing substrates for other kinases, including the Src-family and JAK2 kinases. In this R33 advanced technology development and validation application, we will leverage both of these outcomes to further develop our approach to generate new reagents and protocols for detecting the activity and inhibition of specific kinases or kinase families in intact, live cells. We will also validate the quantitative and multiplexing capabilities of this technology in larger panels of patient samples, collected in collaboration with the Indiana University Simon Cancer Center Hematological Malignancies Tissue Bank. In Aim 1, we plan to address throughput and standardization by developing an isotope-coding strategy that will allow analyses of the same biosensor to be performed in different cells (controls, multiple patient samples) in parallel, followed by pooling and simultaneous analysis. In Aim 2, we will expand the biosensor and mass spectrometry analysis toolbox to include other kinases and kinase families that are important to CML drug resistance and/or other aspects of leukemia biology. In Aim 3, we will validate the application of this technology to patient cells, including optimization of biostability and delivery. This advanced development will establish the quantitative utility of the methodology, and expand the applicability of this technology to kinases and inhibitors relevant beyond CML alone. Ultimately, this work could transform cancer treatment by enabling physicians and patients to make informed decisions about inhibitor treatment choice, monitor efficacy in real time, and catch treatment failure in time to intervene before resistance and relapse become a problem.

描述（由申请人提供）：激酶抑制剂药物已经改变了化疗的面貌，预计仍将是白血病治疗的主要焦点。然而，它们的长期疗效是可变的，这可以归因于它们对目标的活性在患者之间的差异，这种差异可以在开始治疗一周后观察到。我们计划验证一项技术，该技术可以在治疗一开始就量化疗效，使医生和患者有机会立即监测个人的反应和成功的可能性，而不是依赖于目前的观察和等待方法。在帕克实验室获得IMAT R21奖的支持下，我们已经建立了分析慢性髓性白血病（CML）中Bcr-Abl激酶活性和抑制的概念验证。耶鲁大学土耳其实验室还获得了IMAT奖，以支持开发其他激酶（包括src家族和JAK2激酶）底物的概念验证合作工作。在这项R33先进技术开发和验证应用中，我们将利用这些结果进一步开发我们的方法，以产生新的试剂和方案，用于检测完整活细胞中特定激酶或激酶家族的活性和抑制。我们还将与印第安纳大学西蒙癌症中心血液学恶性肿瘤组织库合作，在更大的患者样本中验证该技术的定量和多路复用能力。在目标1中，我们计划通过开发一种同位素编码策略来解决吞吐量和标准化问题，该策略将允许在不同细胞（对照，多个患者样本）中并行执行相同生物传感器的分析，然后进行池化和同时分析。在Aim 2中，我们将扩展生物传感器和质谱分析工具箱，包括其他激酶和激酶家族，这些激酶和激酶家族对CML耐药性和/或白血病生物学的其他方面很重要。在Aim 3中，我们将验证该技术在患者细胞中的应用，包括生物稳定性和递送的优化。这一先进的发展将建立该方法的定量效用，并将该技术扩展到CML以外的激酶和抑制剂的适用性。最终，这项工作可以通过使医生和患者对抑制剂治疗选择做出明智的决定，实时监测疗效，及时发现治疗失败，在耐药性和复发成为问题之前进行干预，从而改变癌症治疗。