Quantitative Mass Spectrometry Assays to Detect Multiple Myeloma and Assess Relap

描述(由申请人提供)：治疗方案和临床干预是由持续的患者监测获得的信息驱动的。目前的蛋白质生物标记物检测方法是基于成熟的技术，包括凝胶电泳法和基于抗体的检测。目前的技术在改善从患者样本中获得的信息方面表现出了突出的前景；这些好处还可以伴随着必须从患者那里收集的材料数量的减少。在这些新兴的方法中，最主要的是定量质谱学。在研究药物传递和分布的反应监测技术的基础上，可以开发分析方法来量化血液或尿液等复杂混合物中的蛋白质生物标记物。这些分析的发展依赖于对特定多肽的选择性检测，这些多肽是蛋白质的酶切片段。多肽被用作替代，以量化其来源的蛋白质。使用三重四极杆质谱仪，完整的多肽可以按质量过滤出来，并通过与背景气体分子(如Ar)的碰撞而碎裂；然后，选择性碎片离子被过滤出来进行检测。这种两级质谱学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。结合反相液相色谱分离，这对完整的相对分子质量和碎片离子质量被称为过渡，提供了三个化学特征来从血液或尿液等复杂基质中分离目标分子的信号。定量质谱学可以用来测量每个样本中的大量蛋白质生物标志物，使得能够在同一样本中对大量临床相关目标进行平行评估。在这项建议中，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，在患者身上产生几种症状。最具破坏性的是骨骼损伤，但这些细胞产生的大量抗体也可以影响血液和肾脏功能中的蛋白质水平的调节。恶性浆细胞分泌大量的免疫球蛋白，作为疾病诊断和病情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法进行监测；这些测量被用作确定患者治疗方案的系统的一部分。定量质谱学方法是对当前临床技术的补充，并可能最终取代这些技术，因为它们提供了关于骨髓瘤患者公共卫生状况的更灵敏的分析和更全面的信息。目标1：本研究将检验从患者采集的血液样本中检测多发性骨髓瘤的改进情况。确定疾病复发将提供当前最高的临床影响；基于新技术的更好的敏感性分析可以为医生提供更多信息，准确指导患者治疗。目的2：改进多发性骨髓瘤患者的尿液采样将使其能够同时评估疾病和相关的肾脏损害。总而言之，这些目标还将使定量质谱学测试与当前的临床分析相比较，说明在患者护理中实施这项新技术的好处。

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.