Quantitative Mass Spectrometry Assays to Detect Multiple Myeloma and Assess Relap

检测多发性骨髓瘤并评估复发的定量质谱分析

• 批准号: 7712397
• 负责人: John M Koomen
• 金额: $ 22.03万
• 依托单位: H. LEE MOFFITT CANCER CTR & RES INST
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-06-01 至 2011-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7712397
• 关键词: Antibodies; Antibody-Producing Cells; Area; Argon; Biological; Biological Assay; Biological Markers; Blood; Blood specimen; Bone Marrow; Cataloging; Catalogs; Characteristics; Chemicals; Cleaved cell; Clinic; Clinical; Clinical Chemistry; Collection; Complex; Complex Mixtures; Coupled; Detection; Development; Diabetes Mellitus; Diagnostic; Disease; Drug Delivery Systems; Drug Kinetics; Electrophoresis; Evaluation; Gases; Hour; Immunoglobulins; Intervention; Ions; Kidney; Kidney Diseases; Lesion; Light; Liquid Chromatography; Literature; Malignant - descriptor; Mass Spectrum Analysis; Measurable; Measurement; Measures; Methods; Molecular Weight; Monitor; Multiple Myeloma; Outcome; Patient Care; Patient Monitoring; Patients; Peptides; Pharmacodynamics; Phase; Physicians; Plasma Cells; Principal Investigator; Process; Proliferating; Proteins; Proteinuria; Proteome; Proteomics; Reaction; Recurrent disease; Regulation; Renal function; Research; Role; Sampling; Serum; Serum Proteins; Severities; Shotgun Sequencing; Signal Transduction; Staging; Symptoms; System; Techniques; Technology; Testing; Treatment Protocols; Urine; assay development; base; bone; clinically relevant; disease diagnosis; gel electrophoresis; improved; internal control; mass spectrometer; multiple reaction monitoring; new technology; numb protein; patient population; prognostic; programs; public health relevance; urinary

DESCRIPTION (provided by applicant): Treatment regimens and clinical intervention are driven by information obtained by ongoing patient monitoring. Current methods for protein biomarker detection are based on well established techniques, including gel electrophoresis and antibody-based detection. Current technologies show outstanding promise for improving the information obtained from patient samples; these benefits can also be coupled with a decrease in the amount of material that has to be collected from the patient. Chief among these emerging methods is quantitative mass spectrometry. Building on reaction monitoring techniques that have been developed to study drug delivery and distribution, assays can be developed to quantify protein biomarkers from complex mixtures, such as blood or urine. The development of these assays relies on selective detection of specific peptides, which are enzymatically cleaved segments of a protein. Peptides are used as a surrogate to quantify their protein of origin. Using a triple quadrupole mass spectrometer, the intact peptide can be filtered out by mass and fragmented by collisions with background gas molecules (e.g. argon); then, selective fragment ions are filtered out for detection. This two-stage mass spectrometry selection enables focused detection of molecules that have the same intact molecular weight and structural fragments, in this case, similar peptide composition and sequence. Coupled with reverse phase liquid chromatography separations, these pairs of intact molecular weight and fragment ion mass, known as transitions, provide three chemical characteristics to isolate the signal of the target molecule from a complex matrix like blood or urine. Quantitative mass spectrometry can be used to measure a large number of protein biomarkers in each sample, enabling evaluation of large panels of clinically relevant targets to be assessed in parallel in the same sample. In this proposal, mass spectrometry assays are developed to detect and quantify immunoglobulins from multiple myeloma patients, which will be compared with the current clinical techniques. In myeloma, malignant plasma cells proliferate in the bone marrow, creating several symptoms in the patient. The most damaging are lesions of the bone, but the large amounts of antibodies produced by these cells can also effect regulation of protein levels in the blood and kidney function. The malignant plasma cells secrete large quantities of immunoglobulins, which serve as biomarkers for disease diagnosis and severity. In the clinic, these proteins are monitored by electrophoresis in blood serum and urine; these measurements are used as part of the system for determining treatment regimens for the patients. Quantitative mass spectrometry methods supplement, and may eventually replace, these current clinical techniques, because they provide more sensitive analysis and more comprehensive information about the condition of the myeloma patient PUBLIC HEALTH RELEVANCE Aim 1: This research will examine improvements in detection of multiple myeloma in blood samples collected from patients. Determination of disease relapse will provide the highest current clinical impact; better sensitivity assays based on newer technology can provide more information to the physician, accurately directing patient treatment. Aim 2: Improvements in sampling the urine of multiple myeloma patients will enable simultaneous assessment of the disease and related kidney damage. Together, these aims will also enable comparison of quantitative mass spectrometry tests to current clinical assays, illustrating the benefits of implementing this new technology in patient care.

描述（由申请人提供）：治疗方案和临床干预措施由持续的患者监测获得的信息驱动。当前用于蛋白质生物标志物检测的方法基于建立的技术，包括凝胶电泳和基于抗体的检测。当前的技术在改善从患者样本获得的信息方面表现出了杰出的希望；这些好处也可以与必须从患者那里收集的材料量减少相结合。这些新兴方法中的主要是定量质谱法。基于已开发用于研究药物输送和分布的反应监测技术，可以开发测定方法来量化复杂混合物（例如血液或尿液）的蛋白质生物标志物。这些测定法的开发依赖于对特定肽的选择性检测，这些肽是蛋白质的酶裂片片段。肽被用作替代其原籍蛋白的替代物。使用三极四极质谱仪，可以通过质量过滤完整的肽，并与背景气体分子（例如氩气）碰撞。然后，将选择性片段离子过滤出来以进行检测。这种两阶段的质谱选择可以重点检测具有相同完整分子量和结构片段的分子，在这种情况下，相似的肽组成和序列。结合反相液相色谱分离，这些完整的分子量重量和碎片离子质量（称为跃迁）提供了三种化学特性，可将靶分子的信号与复杂基质（如血液或尿液）（如血液或尿液）分离。定量质谱法可用于测量每个样品中的大量蛋白质生物标志物，从而评估可以在同一样品中并行评估的大型临床相关靶标。在此提案中，开发了质谱分析，以检测和量化来自多个骨髓瘤患者的免疫球蛋白，这将与当前的临床技术进行比较。在骨髓瘤中，恶性血浆细胞在骨髓中增殖，在患者中产生了几种症状。最具破坏力的是骨骼的病变，但是这些细胞产生的大量抗体也可以影响血液和肾功能中蛋白质水平的调节。恶性血浆细胞分泌大量的免疫球蛋白，这些免疫球蛋白是疾病诊断和严重程度的生物标志物。在诊所中，这些蛋白质通过血清和尿液中的电泳监测。这些测量值用作系统确定患者治疗方案的一部分。定量质谱法补充了这些当前的临床技术，并最终可能取代这些临床技术，因为它们提供了更敏感的分析和有关骨髓瘤患者公共健康相关性状况的更全面的信息目标：这项研究将研究从患者收集的血液样本中检测到多发性骨髓瘤的改善。疾病复发的确定将提供当前最高的临床影响；基于新技术的更好的敏感性测定可以为医师提供更多信息，从而准确指导患者治疗。 AIM 2：改善对多发性骨髓瘤患者的尿液进行取样，将同时评估该疾病和相关的肾脏损伤。这些目标共同将定量质谱测试与当前的临床测定法进行比较，这说明了在患者护理中实施这项新技术的好处。