Optimization of enrichment for MALDI TOF/TOF identification of cancer biomarkers

癌症生物标志物 MALDI TOF/TOF 鉴定的富集优化

• 批准号: 7151111
• 负责人: RADOSLAV GOLDMAN
• 金额: $ 7.76万
• 依托单位: GEORGETOWN UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-09-13 至 2008-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7151111
• 关键词: biomarker; clinical research; early diagnosis; infection; magnetism; mass spectrometry; neoplasm /cancer; peptides; serum

DESCRIPTION (provided by applicant): Early detection of cancer and precancerous conditions improves patient survival. This study optimizes methods for discovery of a diagnostic set of peptides and proteins (P/P) in serum of patients with hepatocellular carcinoma (HCC). To identify biomarkers of HCC, we developed enrichment of low molecular weight (LMW) fraction of serum for matrix assisted laser desorption ionization-time of flight (MALDI- TOF/TOF) mass spectrometry (MS). This method allows high-throughput population screening as well as identification of the peptides of interest by TOF/TOF sequencing. We tested the biomarker discovery method on a pilot set of HCC cases and matched controls, which identified a set of P/P predictive of HCC with greater than 90% prediction accuracy. In this study, we propose to adjust the method to complementary enrichment using WCX and Cu-IMAC magnetic beads, expand the study to include a comparison with premalignant liver disease (150 cases, 150 matched controls, 50 cirrhosis), and to begin identification of the biomarker-candidates by sequencing. The samples come from our ongoing case-control study of HCC in Egypt, a country with an epidemic of hepatitis C infection and HCC. Our goal is to identify biomarkers that would improved early detection of HCC and track the natural progression of chronic viral infection to cancer. The identity of the biomarkers will be verified by MALDI-TOF/TOF sequencing and complementary liquid chromatography/mass spectrometry methods. Defining clinically applicable biomarkers of early-stage cancer has potentially far-reaching consequences for disease management and patient health. The biomarkers could be used to screen high risk populations for early signs of disease; to design and test improved chemoprevention strategies; and to follow disease progression after treatment. Identification of the P/P is expected to generate new hypotheses for targeted disease management.

描述（由申请人提供）：早期发现癌症和癌前病变可提高患者生存率。本研究优化了发现肝细胞癌（HCC）患者血清中诊断性肽和蛋白质（P/P）的方法。为了鉴定HCC的生物标志物，我们开发了用于基质辅助激光解吸电离飞行时间（MALDI-TOF/TOF）质谱（MS）的血清低分子量（LMW）组分的富集。该方法允许高通量群体筛选以及通过TOF/TOF测序鉴定感兴趣的肽。我们在一组试点HCC病例和匹配的对照组上测试了生物标志物发现方法，该方法确定了一组预测HCC的P/P，预测准确率大于90%。在这项研究中，我们建议使用WCX和Cu-IMAC磁珠将该方法调整为互补富集，将研究扩展到包括与癌前肝病（150例，150例匹配对照，50例肝硬化）的比较，并通过测序开始鉴定生物标志物候选物。这些样本来自我们在埃及进行的HCC病例对照研究，埃及是一个丙型肝炎感染和HCC流行的国家。我们的目标是确定生物标志物，以改善HCC的早期检测，并跟踪慢性病毒感染到癌症的自然进展。将通过MALDI-TOF/TOF测序和互补液相色谱/质谱法验证生物标志物的身份。定义早期癌症的临床适用生物标志物对疾病管理和患者健康具有潜在的深远影响。这些生物标志物可用于筛查高危人群的早期疾病体征;设计和测试改进的化学预防策略;以及在治疗后跟踪疾病进展。P/P的鉴定有望为靶向疾病管理产生新的假设。