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由持续患者监测获得的信息驱动。目前用于蛋白质生物标志物检测的方法基于完善的技术，包括凝胶电泳和基于抗体的检测。目前的技术显示出改善从患者样本中获得的信息的突出前景;这些益处还可以与必须从患者收集的材料量的减少相结合。这些新兴方法中最主要的是定量质谱法。基于已经开发用于研究药物递送和分布的反应监测技术，可以开发测定来量化来自复杂混合物（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特异性肽的选择性检测，所述肽是蛋白质的酶促裂解片段。肽被用作替代物来定量其来源蛋白质。使用三重四极杆质谱仪，可以通过质量过滤出完整的肽，并通过与背景气体分子（例如氩气）的碰撞进行碎片化;然后，过滤出选择性碎片离子用于检测。这种两阶段质谱选择能够集中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，具有相似的肽组成和序列。与反相液相色谱分离相结合，这些完整分子量和碎片离子质量对（称为跃迁）提供了三种化学特征，以从复杂基质（如血液或尿液）中分离目标分子的信号。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，从而能够在同一样品中平行评估大量临床相关靶标。在该提案中，开发了质谱分析来检测和定量多发性骨髓瘤患者的免疫球蛋白，并将其与当前的临床技术进行比较。在骨髓瘤中，恶性浆细胞在骨髓中增殖，在患者中产生几种症状。最具破坏性的是骨骼病变，但这些细胞产生的大量抗体也会影响血液和肾脏功能中蛋白质水平的调节。恶性浆细胞分泌大量的免疫球蛋白，作为疾病诊断和严重程度的生物标志物。在临床上，这些蛋白质通过血清和尿液中的电泳进行监测;这些测量结果用作确定患者治疗方案的系统的一部分。定量质谱法补充，并可能最终取代，这些目前的临床技术，因为他们提供了更灵敏的分析和更全面的信息，骨髓瘤患者的条件公共卫生相关目标1：这项研究将检查改善多发性骨髓瘤的血液样本中收集的患者的检测。确定疾病复发将提供最高的当前临床影响;基于更新技术的更好的灵敏度测定可以为医生提供更多信息，准确指导患者治疗。目的2：改善多发性骨髓瘤患者的尿液采样将能够同时评估疾病和相关的肾损害。总之，这些目标还将使定量质谱测试与当前的临床检测进行比较，说明在患者护理中实施这项新技术的好处。