Nanofluidic Wet-Cell for Genomic Analysis of Single Circulating Tumor Cells via Electron Microscopy

通过电子显微镜对单个循环肿瘤细胞进行基因组分析的纳流体湿细胞

• 批准号: 478454-2015
• 负责人: Reisner, Walter
• 金额: $ 4.31万
• 依托单位: McGill University
• 依托单位国家: 加拿大
• 项目类别: Collaborative Health Research Projects
• 财政年份: 2015
• 资助国家: 加拿大
• 起止时间: 2015-01-01 至 2016-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=579520
• 关键词: Nanofluidic; Wet; Cell; Genomic; Analysis

Intratumor genetic heterogeneity is a fundamental challenge in cancer treatment. Genomic variation can occur spatially within a given tumor and between metastatic sub-populations established in different regions of the body. In addition, cancer evolves in time in response to therapy. In order to choose the most effective therapy, there is need for technologies that can yield a comprehensive picture of the "genomic landscape" of a patient's cancer and a measure of how this landscape is evolving. While repeated tissue biopsies are impractical, and provide only local information, circulating tumor cells (CTCs, cancer cells that have entered the bloodstream) may provide a minimally invasive picture of a patient's cancer status. CTCs, however, are both rare and necessarily heterogeneous: CTC-based genomic analysis must thus necessarily be performed at the single cell level. In this proposal we develop a novel approach for direct analysis of single-genomes extracted from single-CTCs at the single-molecule level. Our approach is based on developing a nanofluidic wet-cell that can be operated inside an electron microscope. The wet-cell, which is completely sealed, will permit cycling of DNA through nanochannel arrays positioned below an electronically transparent membrane (formed from either thin silicon nitride or atomically thin graphene sheets). The DNA molecules (specifically labeled with conductive as opposed to fluorescent tags) will be extended in the channels and then mapped via electron microscopy. The approach will yield a comprehensive highresolution map of the genome of a single CTC.

肿瘤内遗传异质性是癌症治疗中的基本挑战。基因组变异可以在空间上发生 在给定的肿瘤内和在身体不同区域建立的转移性亚群之间。此外，癌症 会随着治疗的进行而变化为了选择最有效的疗法，需要能够 产生患者癌症的“基因组景观”的全面图片，并衡量这种景观是如何 进化虽然重复的组织活检是不切实际的，并且仅提供局部信息，但循环肿瘤细胞（CTC， 已经进入血流的癌细胞）可以提供患者癌症状态的微创图片。 然而，CTC是罕见的并且必然是异质的：因此基于CTC的基因组分析必然是不可避免的。 在单细胞水平上进行。在这个建议中，我们开发了一种新的方法，直接分析单基因组 在单分子水平上从单CTC中提取。我们的方法是基于开发一种纳米流体湿细胞， 可以在电子显微镜内操作。湿细胞，这是完全密封的，将允许循环的DNA通过 纳米通道阵列位于电子透明膜（由薄氮化硅或氮化硅形成）下方， 原子级薄的石墨烯片）。DNA分子（用导电标记而不是荧光标记） 将在通道中延伸，然后通过电子显微镜进行映射。该方法将产生全面的高分辨率 单个CTC的基因组图谱。