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: 7955951
• 负责人: GIAMPAOLO MERLINI
• 金额: $ 1.16万
• 依托单位: BOSTON UNIVERSITY MEDICAL CAMPUS
• 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 2010-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7955951
• 关键词: Abdomen; Adipose tissue; Amyloid; Amyloid deposition; Amyloidosis; Apolipoprotein E; Aspirate substance; Biology; 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; Mass Spectrum Analysis; Medicine; 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.

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