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.

该提案的目的是开发基于肽的技术来捕获和扩展体液中的癌细胞，特别是胸膜积液。胸腔积液是与恶性肿瘤有关的常见并发症，尤其是肺癌。通过常规细胞学方法检测胸膜液中癌细胞的敏感性通常很低。人体液中癌细胞的检测通常受到少量恶性细胞的限制，这些恶性细胞被间皮细胞，正常血细胞，成纤维细胞，微生物和细胞碎屑所掩盖。在我们的实验室中，我们使用了由共同研究器Kit Lam博士发明的“单蛋白一面肽”组合库的技术确定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敏感且特异性的细胞生长测定，用于筛选数百万肽珠，用于使用上皮癌细胞的特定肽基序可以附着并生长。使用这种方法，我们已经确定了针对人类恶性细胞系的细胞表面受体（例如非小细胞肺癌的细胞表面受体）的特异性基序。迄今为止，我们已经确定了两个对肺癌细胞高亲和力的特定六肽。使用与这些肽结合的珠，试验研究表明，在大量的红细胞和细胞碎屑的情况下，从胸膜流体中分离出具有可证明的恶性细胞的上皮细胞的可行性。我们假设可以使用与我们迄今为止已经鉴定出的两个特定的六肽配体的珠子，可以通过细胞生长测定的细胞生长测定来分离，富集和保存来自胸膜液的恶性细胞。该提案的具体目的被表述为细胞：1。在大规模合成载有这两个特定六肽肽基序之一的珠子； 2。使用这些肽珠优化细胞生长的孔内测定条件，从胸膜液中捕获和扩展上皮细胞； 3。开发方法来保存和鉴定从细胞生长珠测定法中富含的恶性细胞； 4。开发一种高产量和高通量技术的磁性肽珠技术，用于从胸膜液中分离和富集恶性细胞。 该技术的开发将大大增强我们诊断恶性胸腔积液的能力，并将为捕获和扩展其他体液中的去角质恶性细胞提供模型。