A Nano-Sensing/Micro-Fluidic System for Circulating Tumor Cells (CTCs) Rapid Detection and Diagnosis/Prognosis
应用于循环肿瘤细胞之快速诊断与预后判断之纳米感测与生医微流体系统
Fangang Tseng（曾繁根）
ABSRACT
Despite the recent advancement of biotechnology and pharmaceutical research, cancers remain the leading cause of human mortality. It is vital to diagnose cancers at an early stage when treatment can dramatically improve prognosis. So far, low-cost and easy to operate devices, which allow efficient isolation and sensitive detection of circulating tumor cells (CTCs) for routine blood screening, remain lacking. This talk will introduce a novel micro fluidic platform which can isolate CTCs from the real blood sample in 30 minutes together with a nano sensing system for rapid CTCs identification and diagnosis: this system includes a high throughput 3D Micro-Dialysis1 and Self Assembled Cell Array (SACA) chip2 to quickly self-assemble cells into a densed monolayer (106 cells/cm2) for rapid staining and in-parallel inspection at high speed (6x106 cells/1 hour for 4 images). The distinguished ratio can reach 1 to 1 billion, recovery rate to 80%, and whole process can be finished in two hours for 4 ml blood sample3. In this research, we have successfully enumerating CTC by self-assembled cell array (SACA) chip system for more than 500 patients with newly diagnosed colorectal cancers. Epithelial cell adhesion molecule positive (EpCAM(+)), cluster of differentiation 45 negative (CD45(-)) cells were isolated and enumerated from 2 mL of peripheral vein blood (PB) and inferior mesenteric vein blood (IMV) samples obtained during surgery. We found that the CTC count in PB but not IMV correlates with disease stages. Neoadjuvant chemotherapy did not lead to decreased CTC count in both types of blood samples. With cutoffs of four CTCs per 2 mL of blood, and serum carcinoembryonic antigen (CEA) level of 5 ng/mL, patients with non-metastatic disease were more likely to experience recurrence if they had high PB CTC count and high serum CEA concentration (odds ratio, 8.9). Our study demonstrates a fast CTCs detecting system and the feasibility of enumerating CTCs with a SACA chip in patients with colorectal cancer. Due to the rapid and gentle process, the on-chip isolated CTCs are still in vital and can be further characterized and cultivated for the identification by using nano-mushroom sensors4,5 for further prognosis by reading the information from cancer specific miRNAs. The cultivated/expanded CTCs on chip can be further used as drug testing targets to screening combinatory drugs for AI assisted precision medicine.
尽管近年来生物技术与药物的相关发展在疾病诊断与治疗上日新月异，但癌症的发生与预后判断仍然高占人类死亡原因的前几名。因此能够早期诊断并且做准确的预后判断对癌症的治疗有相当的重要性。最近十年在液态切片活检的发展上，除了分子等级的检测外，细胞等级的侦检也成为相当重要的依据，因为可以提供更完整与有功能意义的数据来提升癌症诊断与治疗的准确性。血液中循环肿瘤细胞的检测也在近年成为相当重要的依据之一。但如何快速且精确地在血液样本众多的血球细胞中，找到数量仅千万分之一的循环肿瘤细胞仍然是一个困难且重要的议题。因为数量的稀少,因此这些细胞找到之后的单细胞基因体以及蛋白质体的检测与分类，以及放大培养成为药物筛检的工具，仍然是相当困难但重要的议题。本演讲将介绍一个在实验室过去几年所发展的纳米感测微流体系统,其利用细胞自组装特性以及纳米感测材料来针对循环肿瘤细胞(CTCs)做分析、撷取、诊断、以及预后判断。这个细胞阵列自组装系统 (Self Assembled Cell Array, or SACA, chip)可以在五分钟内将血液中所有细胞自组装成为一个单一的高密度细胞层(106cells/cm2)，用来做稀少细胞的搜寻、计数、分类、撷取、染色、以及其基因体或蛋白质体的单细胞检测。目前此系统在过去五年内已完成超过500例的临床病人检体实验，对于大肠直肠癌第三与四期的病人，其两年内癌症复发的预后判断的准确率可以达到五成以上，预测成功与失败的比值可以超过10以上，有相当不错的预后判断能力。此系统未来将搭配纳米检测用于单细胞的基因体与蛋白质体分析,以提供医生癌症化疗或免疫治疗的用药依据。以此系统所筛检出的循环肿瘤细胞在数量扩张后，可以应用于组合性药物的筛检，以供精准医疗使用。
BIOGRAPHY
Dr. Fan-Gag (kevin) Tseng received his Ph.D. degree in mechanical engineering from UCLA, USA, in 1998. He is currently a distinguished professor of ESS Dept. as well as NEMS I., and the Vice President for R&D at NTHU. He was elected an ASME fellow in 2014. His research interests are in the fields of BioNEMS, Biosensors, Micro-Fluidics, Tissue Chips, and Fuel Cells. He received 60 patents, wrote 8 book chapters, published more than SCI 220 Journal papers and 360 conference technical papers. He received several awards, including, Shakelton Scholar, twice National Innovation Award, twice Outstanding in research award, and Mr. Wu, Da-Yo Memorial Award from MOST, Taiwan, and more than twenty best papers and other awards in various international conferences and competitons.
曾繁根博士于1998年毕业于美国洛杉矶加州大学的机械工程学系。他目前是台湾清华大学的特聘教授兼研发长，服务于工程与系统科学系、纳米工程与系统科学研究所、以及前瞻物质科技研究中心，也合聘于中央研究院应用科学中心研究员。他在2014年获选为美国机械工程协会会士。他的研究兴趣在于生医纳微机电系统、微流体系统、生医感测器、组织晶片、以及纳米氢能与燃料电池系统等。他拥有超过60个以上的专利，出版超过220篇期刊论文、360篇国际研讨会论文以及8本以上专书。他曾经得过超过30个以上的奖项，包含两次国家新创奖、科技部沙克尔顿学者奖助、两次科技部杰出研究奖、科技部吴大猷先生纪念奖、有庠科技论文奖、以及东元科技奖等。