Capturing cancer cells with peptide bead technology

利用肽珠技术捕获癌细胞

• 批准号: 6446600
• 负责人: DERICK LAU
• 金额: $ 14.83万
• 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 2004-02-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6446600
• 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.

该提案的目的是开发一种基于肽的技术来捕获和扩增体液（特别是胸腔积液）中的癌细胞。胸腔积液是与恶性肿瘤尤其是肺癌相关的常见并发症。通过常规细胞学方法检测胸水中的癌细胞的灵敏度通常较低。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，我们利用共同研究员Kit Lam博士发明的“一面包一肽”组合库技术，鉴定出对恶性细胞具有高亲和力的肽配体。我们的实验室还开发了一种灵敏且特异性的珠上细胞生长测定法，用于筛选数百万个肽珠的特定肽基序，以利用上皮癌细胞的附着和生长。使用这种方法，我们已经鉴定了对人类恶性细胞系（例如非小细胞肺癌细胞系）的细胞表面受体特异的肽特异性基序。迄今为止，我们已经鉴定出两种对肺癌细胞具有高亲和力的特异性六肽。使用与这些肽缀合的珠子，初步研究证明了从胸水中分离上皮细胞的可行性，其中含有大量红细胞和细胞碎片中已证实的恶性细胞。我们假设，可以通过使用与我们迄今为止已鉴定的两种特定六肽配体缀合的珠子进行珠上细胞生长测定来分离、富集和保存胸水中的恶性细胞。该提案的具体目标被表述为细胞： 1. 大规模合成携带这两种特定六肽基序之一的珠子； 2. 使用这些肽珠捕获并扩增胸膜液中的上皮细胞，优化珠上细胞生长测定的条件； 3. 开发保存和鉴定从细胞珠上生长实验富集的恶性细胞的方法； 4. 开发高产、高通量的磁性肽珠技术，用于从胸水中分离和富集恶性细胞。 该技术的发展将大大提高我们诊断恶性胸腔积液的能力，并为捕获和扩增其他体液中脱落的恶性细胞提供模型。