Peptide Fingerprint multiplex analysis by MS/MS for primary immundeficiency

通过 MS/MS 进行原发性免疫缺陷的肽指纹多重分析

• 批准号: 7774925
• 负责人: Sihoun Hahn
• 金额: $ 29.25万
• 依托单位: SEATTLE CHILDREN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-07-01 至 2012-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7774925
• 关键词: Affect; Area; Binding; Blood; Blood Volume; Brain; CD3 Antigens; Ceruloplasmin; Cessation of life; Characteristics; Chromosomes; Clinic; Clinical; Computer Simulation; Congenital Disorders; Copper; DNA; Data; Daughter; Defect; Detection; Diagnosis; Diagnostic; Diagnostic tests; Digestion; Disease; Disease model; Early Diagnosis; Early identification; Excision; Genetic Recombination; Goals; Hemorrhage; Hepatolenticular Degeneration; Human; Human Genome; Immunologic Deficiency Syndromes; Immunology; Inborn Genetic Diseases; Infection; Ions; Lead; Life; Lung; Mass Spectrum Analysis; Membrane; Menkes Kinky Hair Syndrome; Metabolism; Methodology; Methods; Molecular; Morbidity - disease rate; Neonatal Screening; Organ; Outcome; Paper; Patient Care; Patients; Peptide Mapping; Peptides; Plasma; Plasma Proteins; Process; Productivity; Proteins; Proteomics; Quality of life; Research; Sampling; Screening procedure; Sensitivity and Specificity; Severe Combined Immunodeficiency; Spottings; T-Cell Receptor; T-Cell Receptor Genes; Technology; Testing; Time; Validation; Wisconsin; Wiskott-Aldrich Syndrome; Work; X-Linked Agammaglobulinemia; base; design; early childhood; experience; improved; infancy; innovation; medical attention; meetings; multidisciplinary; population based; prevent; protein aminoacid sequence; protein complex; public health relevance; rapid diagnosis; research study; tandem mass spectrometry; tool

DESCRIPTION (provided by applicant): Severe Combined Immunodeficiency (SCID), Wiskott-Aldrich Syndrome (WAS), and X-Linked Agammaglobulinemia (XLA) are three primary immune deficiency disorders associated with early lethality if not diagnosed and treated appropriately. Early recognition, diagnosis, and preemptive treatment are critical for improving outcomes in these disorders however; there is a fundamental gad in our ability to perform population-based screening for more than just SCID. The long-term goal is to develop a viable multiplex testing methodology that will allow simultaneous identification of multiple congenital disorders characterized by absence of specific proteins using small sample volumes of blood dried onto filter paper. The objective of this particular application is to identify and characterize unique "fingerprints" peptides that can be used to rapidly identify patients with three severe immunodeficiency syndromes that will be used as model disorders: Severe Combined Immunodeficiency (SCID), Wiskott-Aldrich Syndrome (WAS), and X-Linked Agammaglobulinemia (XLA). Dried blood spots will be used as the substrate and the testing approach will be tandem mass spectrometry. The central hypothesis of this application is that tandem mass spectrometry can used to detect the presence or absence of key "fingerprint" peptides derived from CD3, WASP and BTK (markers for SCID, WAS, and XLA respectively). This hypothesis has been formulated based on our preliminary data (deWilde et.al., 2008). The rationale for the proposed research is that rapid, early detection of severe primary immunodeficiency disorder such as SCID, WAS, and XLA will allow for preemptive treatment that will prevent death and morbidity thereby improving quality of life and long-term productivity. The central hypothesis will be tested by pursuing three specific aims: 1.) Identify key "fingerprint" peptides for CD3, WASP, and BTK; marke proteins for SCID, WAS. And XLA respectively. 2.) Define the suitability of each predicted "fingerprint" peptide for use in tandem mass spectrometry (MS/MS) and optimize its detection. 3.) Validate the ability of each "fingerprint" peptide to identify the absence of CD3, WASP, and BTK in dried blood spots from patients with SDIC, WAS, and XLA as well as controls. The approach is innovative because of the new methods we have developed, expanding the use of an existing technology to diagnose congenital disorders that result from absence of a specific protein. These tools have the potential to increase our capacity for early and rapid identification of several severe primary immunodeficiency disorders. The proposed research is significant, because it will provide a testing methodology that is viable both for newborn screening and for rapid diagnostics for these and other severe immunodeficiency and congenital disorders. This will provide opportunities to identify patients early in life and provide life-saving therapies. In addition, this is expected to fundamentally advance the use of proteomic methodologies for diagnostic testing, an area that holds great promise but is largely untapped. PUBLIC HEALTH RELEVANCE: The proposed studies focus on early and rapid diagnosis of the severe, life-threatening immunodeficiency syndromes Severe Combined Immunodeficiency (SCID), Wiskott-Aldrich Syndrome (WAS), and X-Linked Agammaglobulinemia (XLA). This application is designed to utilize an innovative tandem mass spectroscopy approach to quickly diagnose these disorders using blood spots dried onto filter paper, because of the high-throughput, multiplex potential of this method, it will dramatically expand our ability to perform newborn screening and rapid diagnostic testing for these and other disorders using very small blood volumes. This will directly impact patient care by providing early identification and the opportunity for preemptive treatment for life threatening immunodeficiency disorders.

描述（由申请人提供）：严重联合免疫缺陷（SCID）、Wiskott-Aldrich综合征（WAS）和X-Linked Agammaglobulinemia （XLA）是三种原发性免疫缺陷疾病，如果不适当诊断和治疗，会导致早期死亡。然而，早期识别、诊断和预防性治疗对于改善这些疾病的预后至关重要；我们进行基于人群的筛查的能力有一个根本的缺陷，而不仅仅是SCID。长期目标是开发一种可行的多重检测方法，利用在滤纸上干燥的少量血液样本，可以同时识别以缺乏特定蛋白质为特征的多种先天性疾病。这种特殊应用的目的是识别和表征独特的“指纹”肽，可用于快速识别三种严重免疫缺陷综合征患者，这些综合征将用作模型疾病：严重联合免疫缺陷综合征（SCID）， Wiskott-Aldrich综合征（WAS）和X-Linked Agammaglobulinemia （XLA）。干燥的血斑将被用作底物，测试方法将采用串联质谱法。该应用的中心假设是，串联质谱法可用于检测来自CD3、WASP和BTK（分别为SCID、WAS和XLA的标记）的关键“指纹”肽的存在或缺失。这一假设是根据我们的初步数据制定的（deWilde等）。, 2008)。拟议研究的基本原理是，快速、早期发现严重的原发性免疫缺陷障碍，如SCID、WAS和XLA，将允许先发制人的治疗，从而防止死亡和发病率，从而提高生活质量和长期生产力。中心假设将通过追求三个具体目标来检验：鉴定CD3、WASP和BTK的关键“指纹”肽；标记SCID、WAS蛋白。和XLA。2.） 确定每个预测的“指纹”肽在串联质谱（MS/MS）中的适用性，并优化其检测。3.） 验证每个“指纹”肽在SDIC、WAS和XLA患者以及对照的干血斑中识别CD3、WASP和BTK缺失的能力。这种方法是创新的，因为我们开发了新的方法，扩大了现有技术的使用范围，以诊断由于缺乏特定蛋白质而导致的先天性疾病。这些工具有可能提高我们早期和快速识别几种严重的原发性免疫缺陷疾病的能力。拟议的研究意义重大，因为它将为新生儿筛查和快速诊断这些和其他严重免疫缺陷和先天性疾病提供一种可行的测试方法。这将为在生命早期识别患者并提供挽救生命的治疗提供机会。此外，这有望从根本上推动蛋白质组学方法在诊断测试中的应用，这是一个前景广阔但尚未开发的领域。