Capturing cancer cells with peptide bead technology

利用肽珠技术捕获癌细胞

• 批准号: 6622462
• 负责人: DERICK LAU
• 金额: $ 14.85万
• 依托单位: UNIVERSITY OF CALIFORNIA AT DAVIS
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-03-18 至 2005-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6622462
• 关键词: cell adhesion; cell sorting; flow cytometry; high throughput technology; human tissue; lung neoplasms; neoplasm /cancer genetics; neoplastic cell; oncogenes; peptides; pleural cavity effusion; point mutation; polymerase chain reaction; polystyrenes; single strand conformation polymorphism; technology /technique development

The purpose of this proposal is to develop a peptide-based technology to capture and expand cancer cells in body fluids, specifically pleural effusion. Pleural effusion is a common complication associated with malignancies especially lung cancer. The sensitivity is generally low in detecting cancer cells in pleural fluid by conventional cytologic methods. Detection of cancer cells in body fluids is commonly limited by the small number of malignant cells which are obscured by the presence of mesothelial cells, normal blood cells, fibroblasts, microorganisms and cellular debris. In our laboratory, we have identified peptide ligands with high affinity for malignant cells using the technology of 'one-bread one-peptide' combinatorial library invented by the co-investigator, Dr. Kit Lam. A sensitive and specific cell-growth-on-bead assay has also been developed in our laboratory for screening millions of peptide beads for specific peptide motifs to use epithelial cancer cells can attach and grow. Using this method, we have identified peptide specific motifs specific for cell- surface receptors of human malignant cell lines such as that from non- small cell lung cancers. To date, we have identified two specific hexapeptides with high affinity for lung cancer cells. Using beads conjugated with these peptides, pilot studies have demonstrated the feasibility of isolating epithelial cells from pleural fluids with proven malignant cells amid a large number of red blood cells and cell debris. We hypothesize that malignant cells from pleural fluid can be isolated, enriched and preserved by the cell-growth-on-bead assay using beads conjugated with the two specific hexapeptide ligands that we have identified to date. The specific aims of this proposal are formulated as cells: 1. To synthesize, in large scale, beads carrying one of these two specific hexapeptide motifs; 2. To optimize conditions of cell-growth- on-bead assay using these peptide beads to capture and expand epithelial cells from pleural fluid; 3. To develop methods to preserve and identify malignant cells enriched from the cell-growth-on-bead assay; 4. To develop a high-yield and high-throughput technology of magnetic peptide-beads for isolating and enriching malignant cells from pleural fluid. The development of this technology will greatly enhance our ability of diagnosing malignant pleural effusion, and it will provide a model for capturing and expanding exfoliated malignant cells in other body fluids.

这项提议的目的是开发一种基于多肽的技术来捕获和扩大体液中的癌细胞，特别是胸腔积液。胸腔积液是恶性肿瘤尤其是肺癌的常见并发症。常规细胞学方法检测胸水中癌细胞的敏感性普遍较低。体液中癌细胞的检测通常受到少量恶性细胞的限制，这些细胞被间皮细胞、正常血细胞、成纤维细胞、微生物和细胞碎片的存在所掩盖。在我们的实验室里，我们利用共同研究人员林洁仪博士发明的一面包一肽组合文库技术，筛选出了对恶性肿瘤细胞具有高亲和力的多肽配体。我们实验室还开发了一种灵敏和特异的细胞珠上生长试验，用于筛选数百万个特定多肽基序的多肽微珠，以利用上皮性癌细胞可以附着和生长。利用这种方法，我们已经鉴定了人类恶性细胞系(如非小细胞肺癌)细胞表面受体的特异性多肽基序。到目前为止，我们已经鉴定出两个对肺癌细胞具有高亲和力的特异性六肽。使用与这些多肽结合的珠子，初步研究证明了从大量红细胞和细胞碎片中分离出具有已证实的恶性细胞的胸水中的上皮细胞的可行性。我们假设，胸水中的恶性细胞可以通过珠上细胞生长试验分离、浓缩和保存，该试验使用的是与我们迄今鉴定的两种特定六肽配体结合的珠子。这项建议的具体目标是细胞：1.大规模合成携带这两个特定六肽基序之一的小球；2.优化使用这些小球进行细胞生长小球实验的条件，以捕获和扩增胸水中的上皮细胞；3.开发保存和鉴定从细胞小球生长实验中浓缩的恶性细胞的方法；4.开发一种高产率和高通量的磁多肽小球技术，用于分离和浓缩胸水中的恶性细胞。这项技术的发展将极大地提高我们诊断恶性胸腔积液的能力，并将为捕获和扩增其他体液中的脱落恶性细胞提供一个模型。