Phosphoproteomic signatures for early detection and stratification of AML

用于 AML 早期检测和分层的磷酸化蛋白质组学特征

• 批准号: 8893365
• 负责人: Alex Kentsis
• 金额: $ 22.87万
• 依托单位: DANA-FARBER CANCER INST
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-08-01 至 2017-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8893365
• 关键词: Acute Myelocytic Leukemia; Agreement; Biological Assay; Biological Markers; Biology; Blood capillaries; Cancer Center; Cell Survival; Clinical; DNA Sequence; DNA Sequence Alteration; DNA lesion; Data; Detection; Development; Diagnosis; Diagnostic; Disease; Drug Targeting; Early Diagnosis; FLT3 gene; Failure; Foundations; Generations; Genes; Genetic; Genome; Genotype; Goals; Health; Hematopoietic Neoplasms; Human; Individual; Karyotype; Lead; Lesion; Leukemic Cell; Link; Mass Spectrum Analysis; Measures; Mediating; Molecular; Mutation; NPM1 gene; Ohio; Oncogenic; Outcome; Pathway interactions; Patients; Performance; Phenotype; Phosphoproteins; Phosphorylation; Phosphotransferases; Population; Primary Neoplasm; Prognostic Marker; Proteome; Proteomics; Recurrence; Research Proposals; Resistance; Sampling; Secure; Signal Pathway; Signal Transduction; Signaling Molecule; Specimen; Stem cells; Stratification; Techniques; Technology; Time; Tissues; Translating; Translations; Treatment Failure; Validation; Work; base; capillary; cell growth; chemotherapy; clinical application; cohort; disease phenotype; drug efficacy; exome sequencing; genetic analysis; genome-wide; high risk; improved; leukemogenesis; member; nanoimmunoassay; next generation; novel; point of care; prognostic; success; tumor; validation studies

 DESCRIPTION (provided by applicant): Acute myeloid leukemia remains a lethal disease, especially for patients who do not respond to intensive chemotherapy. Despite significant new data resulting from next-generation DNA sequencing techniques, it remains difficult to link specific genetic mutations to the molecular mechanisms that drive aberrant proliferation of leukemic cells. Moreover, some 50% of AML patients who display a normal karyotype and who lack mutations in key genes such as FLT3 or NPM1 are resistant to induction chemotherapy. In contrast to gene sequencing, phosphoproteomic approaches provide a direct and quantitative measure of the signaling molecules that mediate leukemogenesis, circumventing the need to decipher 'driver' mutations from the very large number of non-functional 'passenger' DNA lesions that accumulate in tumors. In preliminary studies for this research proposal, we discovered novel mechanisms by which signaling pathways support AML cell growth and survival. In related work, we have developed DEEP SEQ mass spectrometry, a scalable platform that provides true, genome-wide proteome quantification. Consistent with the objectives described in FOA PA-12-220, which encourages "development and improvement of specific technologies for quantitative detection of novel biomarkers associated with hematopoietic malignancies," with a focus on "early detection, prediction of progression, and recurrence of hematopoietic malignancies, especially in high-risk individuals," we will further improve and utilize our DEEP SEQ platform to identify phosphorylation-based, functional biomarkers in primary tumors of high-risk AML patients. We will validate our phosphoproteomic data using capillary nanoimmunoassays and targeted mass spectrometry, analytical formats suitable for point-of-care clinical applications. Our novel discovery and validation strategy will capture the signaling pathways associated with therapy failure in high-risk AML patients, and lay the foundation for translation of our phosphoproteomic data into improved diagnostics. Our study plan has two specific aims: Specific Aim 1 is to establish phosphoproteomic signatures of chemoresistance and therapy failure in diagnostic specimens from patients with AML. We will use DEEP SEQ mass spectrometry to identify phosphorylation markers associated with chemoresistance and therapy failure. Importantly, we have secured collaborative agreements with colleagues at four major cancer centers to access primary AML patient samples that are well-annotated with respect to genetic background and clinical outcomes. Specific Aim 2 is to validate phosphoproteomic markers and access their prognostic performance in independent cohorts of AML patients. We will validate our phosphoproteomic biomarkers across a larger, independent patient cohort to determine prognostic performance. To facilitate eventual translation to clinical application we will perform these validation studies using the complementary techniques of capillary nanoimmunoassays and targeted mass spectrometry.

 描述（由适用提供）：急性髓细胞性白血病仍然是一种致命的疾病，尤其是对于对强化化学疗法反应的患者。尽管下一代DNA测序技术产生了重大的新数据，但仍然很难将特定的遗传突变与驱动白血病细胞异常增殖的分子机制联系起来。此外，大约有50％的AML患者表现出正常的核型并且缺乏关键基因（例如FLT3或NPM1）的突变具有抗诱导化疗性。与基因测序相反，磷酸蛋白质组学方法提供了介导白血病发生的信号分子的直接和定量度量，从而从大量非功能的“乘客” DNA疾病中解解了“驱动器”突变的需求，这些DNA在肿瘤中积聚。在该研究建议的初步研究中，我们发现了信号通路支持AML细胞生长和生存的新机制。在相关工作中，我们开发了深SEQ质谱，这是一个可扩展的平台，可提供真正的全基因组蛋白质组定量。与FOA PA-12-220中描述的目标一致，该目标鼓励“对与造血性恶性肿瘤相关的新型生物标志物进行定量检测的特定技术的发展和改进，重点是“早期发现，预测，预测的进展，对血液恶性肿瘤的恢复，尤其是在高层型中，我们将识别和识别我们的实质性，我们将改善我们的境地，我们可以改善我们的进步，我们可以进一步改善我们的进步，我们可以进一步改善我们的进步，我们可以进一步改善我们的进步，我们可以进一步改善我们的进步，我们可以进一步改善我们的进一步范围。高危AML患者原发性肿瘤中的功能性生物标志物。我们将使用毛细血管纳米免疫测定和靶向质谱，分析格式验证磷酸蛋白质组学数据，适用于护理临床应用。我们的新颖发现和验证策略将捕获高危AML患者中与治疗失败相关的信号通路，并为将我们的磷蛋白质组学数据转化为改进的诊断剂。我们的研究计划有两个具体的目的：特定目的1是在AML患者的诊断标本中建立化学耐药和治疗失败的磷酸蛋白质组学特征。我们将使用深层SEQ质谱法来鉴定与化学耐药性和治疗衰竭相关的磷酸化标记。重要的是，我们已经与四个主要癌症中心的同事达成了协作协议，以获取有关遗传背景和临床结果井井有条的初级AML患者样本。具体目的2是验证磷酸蛋白质组学标记并在独立的AML患者中获得预后性能。我们将在更大的独立患者队列中验证我们的磷酸蛋白质组学生物标志物，以确定预后性能。为了促进最终转换为临床应用，我们将使用毛细管纳米免疫测定和靶向质谱的完整技术进行这些验证研究。