Rapid unbiased isolation and in situ RNA analysis of circulating tumor cells using a magnetic micropore-based diagnostic chip

使用基于磁性微孔的诊断芯片对循环肿瘤细胞进行快速无偏分离和原位 RNA 分析

• 批准号: 9277638
• 负责人: David Aaron Issadore
• 金额: $ 38.16万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-09-20 至 2020-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9277638
• 关键词: Address; Algorithmic Analysis; Benchmarking; Benign; Biological Assay; Biological Markers; Biology; Biopsy; Blood; Blood Platelets; Blood Volume; Blood specimen; Body part; Cancer Etiology; Cells; Cessation of life; Clinical; Computer software; Cultured Cells; Cultured Tumor Cells; Custom; Data; Detection; Development; Devices; Diagnosis; Diagnostic; Disease; Disease Progression; Drug Targeting; Elements; Epithelial; Erythrocytes; Fluorescent in Situ Hybridization; Genetic Engineering; Genetic Transcription; Goals; Gold; Histology; Hour; Human; Hybrids; Image; Image Analysis; In Situ; Individual; Italy; Label; Lead; Lesion; Leukocytes; Magnetism; Malignant - descriptor; Malignant Neoplasms; Malignant neoplasm of pancreas; Measurement; Measures; Methods; Microfluidics; Modeling; Molecular; Molecular Profiling; Monitor; Mus; Neoplasm Circulating Cells; Neoplasm Metastasis; Onset of illness; Outcome; Pancreas; Patient-Focused Outcomes; Patients; Performance; Primary Neoplasm; Procedures; Process; Publishing; RNA; RNA Probes; RNA Sequences; RNA analysis; Sampling; Series; Sorting - Cell Movement; Specificity; Stream; Survival Rate; Suspensions; Techniques; Technology; Testing; Time; Tracer; United States; Whole Blood; base; cancer biomarkers; design; drug efficacy; experimental study; improved; innovation; iterative design; minimally invasive; molecular marker; mouse model; nanomaterials; next generation; non-invasive monitor; noninvasive diagnosis; novel; novel strategies; pancreatic cancer cells; personalized medicine; single molecule; tool; transcriptome sequencing

ABSTRACT: Pancreatic cancer is the fourth most common cause of cancer related death in the United States, with a five year survival rate of only 4%. We have recently shown that circulating pancreatic cells (CPCs) can be detected in the blood at the onset of the disease cycle in both mice and humans. These findings present an opportunity to develop a non-invasive diagnostic for pancreatic cancer that has enormous potential to improve patient outcomes. The current gold-standard for circulating tumor cell (CTC) detection fails to measure the extremely sparse and heterogeneous CPCs. Even next generation microfluidic technologies, which do not rely on epithelial markers, have limited utility due to the inherently low-throughput of microfluidics and the large sample volumes of blood (V > 10 mL) necessary for ultra-rare cell detection. To address these challenges, we are developing a new approach to rare cell detection, using magnetic micropores, which combines the benefits of micro-scale sorting with extremely fast flow rates (100x faster than typical microfluidic approaches2,5-7). In addition to its improved throughput, our technique is robust against unprocessed clinical samples, allowing rare CPCs and CPC clusters to be isolated directly from whole blood. Moreover, we are integrating this approach with ultra-rapid on-chip RNA fluorescence in-situ hybridization (RNA FISH), enabling single molecule, multiplexed RNA analysis in individual rare cells. Our device, which combines the above features into a self-contained, automated format, is designed to extract the RNA expression of single CPCs in clinical settings.

抽象的： 胰腺癌是美国癌症相关死亡的第四大常见原因 美国的五年生存率仅为 4%。我们最近表明，流通 在两种疾病的疾病周期开始时，都可以在血液中检测到胰腺细胞（CPC） 老鼠和人类。这些发现为开发非侵入性诊断提供了机会 对于胰腺癌来说，它具有改善患者预后的巨大潜力。目前的 循环肿瘤细胞（CTC）检测的金标准无法测量极其稀疏的细胞 和异构 CPC。即使是下一代微流体技术，也不依赖于 由于微流体固有的低通量和上皮标记物的实用性有限 超稀有细胞检测所需的大量血液样本 (V > 10 mL)。到 为了应对这些挑战，我们正在开发一种新的稀有细胞检测方法，使用 磁性微孔，结合了微尺度分选和极快速的优点 流速（比典型微流体方法快 100 倍2,5-7）。除了其改进的 吞吐量，我们的技术对未处理的临床样本具有稳健性，允许稀有的 CPC 和CPC簇直接从全血中分离。此外，我们正在整合这个 超快速芯片上 RNA 荧光原位杂交 (RNA FISH) 方法，使 对单个稀有细胞进行单分子、多重 RNA 分析。我们的设备结合了 将上述功能转化为独立的、自动化的格式，旨在提取 RNA 临床环境中单个 CPC 的表达。