PROTEOMIC APPROACH TO THE STUDY OF SYSTEMIC AMYLOIDOSIS

系统性淀粉样变性研究的蛋白质组学方法

• 批准号: 8170917
• 负责人: GIAMPAOLO MERLINI
• 金额: $ 1.13万
• 依托单位: BOSTON UNIVERSITY MEDICAL CAMPUS
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-06-01 至 2011-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8170917
• 关键词: Abdomen; Adipose tissue; Amyloid; Amyloid deposition; Amyloidosis; Apolipoprotein E; Aspirate substance; Computer Retrieval of Information on Scientific Projects Database; Crystallins; Deposition; Diagnosis; Disease; Down-Regulation; E protein; Employee Strikes; Fatty acid glycerol esters; Fine needle aspiration biopsy; Fingerprint; Functional disorder; Funding; Gel; Grant; Infiltration; Institution; Length; Light-Chain Immunoglobulins; Methodology; Mutation; Organ; Oxidoreductase; Patients; Pattern; Peptides; Post-Translational Protein Processing; Prealbumin; Protein Isoforms; Proteins; Proteome; Proteomics; Research; Research Personnel; Resources; Sampling; Serum; Source; Specimen; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Spottings; Therapeutic; Two-Dimensional Polyacrylamide Gel Electrophoresis; United States National Institutes of Health; Variant; design; insight; novel; peroxiredoxin; subcutaneous; two-dimensional

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. In systemic amyloidoses, widespread deposition of protein as amyloid causes severe organ dysfunction. It is necessary to discriminate among the different forms of amyloid to design an appropriate therapeutic strategy. We developed a proteomics methodology utilizing two-dimensional polyacrylamide gel electrophoresis followed by matrix-assisted laser desorption/ionization mass spectrometry and peptide mass fingerprinting to directly characterize amyloid deposits in abdominal subcutaneous fat obtained by fine needle aspiration from patients diagnosed as having amyloidoses typed as immunoglobulin light chain or transthyretin. Striking differences in the two-dimensional gel proteomes of adipose tissue were observed between controls and patients and between the two types of patients with distinct, additional spots present in the patient specimens that could be assigned as the amyloidogenic proteins in full-length and truncated forms. In patients heterozygotic for transthyretin mutations, wild-type peptides and peptides containing amyloidogenic transthyretin variants were isolated in roughly equal amounts from the same protein spots, indicative of incorporation of both species into the deposits. Furthermore novel spots unrelated to the amyloidogenic proteins appeared in patient samples; some of these were identified as isoforms of serum amyloid P and apolipoprotein E, proteins that have been described previously to be associated with amyloid deposits. Finally changes in the normal expression pattern of resident adipose proteins, such as down-regulation of alphaB-crystallin, peroxiredoxin 6, and aldo-keto reductase I, were observed in apparent association with the presence of amyloid, although their levels did not strictly correlate with the grade of amyloid deposition. This proteomics approach not only provides a way to detect and unambiguously type the deposits in abdominal subcutaneous fat aspirates from patients with amyloidoses but it may also have the capability to generate new insights into the mechanism of the diseases by identifying novel proteins or protein post-translational modifications associated with amyloid infiltration. During the current year, preliminary results for a proposal were prepared and the proposal was submitted, together with other centers, that proposed an expansion of this project, but it has not yet been funded.

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 在系统性淀粉样变性中，蛋白质作为淀粉样蛋白广泛沉积导致严重的器官功能障碍。有必要区分不同形式的淀粉样蛋白，以设计适当的治疗策略。我们开发了一种蛋白质组学方法，利用二维聚丙烯酰胺凝胶电泳，然后基质辅助激光解吸/电离质谱和肽质量指纹直接表征淀粉样蛋白沉积在腹部皮下脂肪获得细针穿刺从诊断为淀粉样蛋白病分型为免疫球蛋白轻链或甲状腺素运载蛋白的患者。在对照组和患者之间以及两种类型的患者之间观察到脂肪组织的二维凝胶蛋白质组的显著差异，患者标本中存在不同的额外斑点，这些斑点可以被分配为全长和截短形式的淀粉样蛋白。在甲状腺素运载蛋白突变杂合子的患者中，从相同的蛋白质斑点中分离出大致等量的野生型肽和含有淀粉样蛋白生成性甲状腺素运载蛋白变体的肽，这表明两种物质均掺入沉积物中。此外，在患者样本中出现了与淀粉样蛋白无关的新斑点;其中一些被鉴定为血清淀粉样蛋白P和载脂蛋白E的同种型，这些蛋白质先前已被描述为与淀粉样蛋白沉积相关。最后，观察到居民脂肪蛋白的正常表达模式的变化，如α B-晶状体蛋白、过氧化物氧还蛋白6和醛酮还原酶I的下调，与淀粉样蛋白的存在明显相关，尽管它们的水平与淀粉样蛋白沉积的程度并不严格相关。这种蛋白质组学方法不仅提供了一种方法来检测和明确类型的沉积物在腹部皮下脂肪吸出物从患者淀粉样蛋白，但它也可能有能力产生新的见解的疾病的机制，通过识别新的蛋白质或蛋白质翻译后修饰与淀粉样蛋白浸润。在本年度，拟定了一项提案的初步结果，并与其他中心一起提交了该提案，建议扩大该项目，但尚未获得资金。