肿瘤血行微转移的传感预警与即时监测技术的研究

Tumor blood-way micrometastasis is the initial stage of tumor cells invading into the blood circulation, where the formed tumors are too minuscule to be detected clinically by classic analysis technologies. Its accurate diagnostics and early warning, and point-of-care (POC) monitoring is of significance for the control of micrometastasis, so as to increase the tumor curative rate and improve the therapy efficiency of radiotherapy, chemotherapy and surgery for enhanced survival rate of the patients. In this project, a series of sensitive, selective and efficient biosensing platforms are systematically established for the detection of direct micrometastasis biomarkers in blood, circulating tumor cells (CTC), aiming at early warning of tumor micrometastasis. A microsieve device is fabricated to screen CTC combining with magnetic separation technique using "positive" and "negative" magnetic bio-beads to capture CTC. Some isothermal gene amplification techniques will be introduced to achieve real-time reverse transcription and one-step amplification of targeting genes. Also, the synergetic effects of tool enzymes of nucleic acids and the signal-amplification strategies will be employed. The biosensing platforms can allow for the ultra-sensitive detection of rare CTC in blood toward accurate early warning of clinical tumor micrometastasis. Moreover, new analysis methodologies are developed for POC monitoring of tumor micrometastasis during its therapies of radiotherapy, chemotherapy and surgery by way of simultaneous detection of different tumor biomarkers, i.e., tumor-specific proteins and tumor cell-free genes. A lab-made microarray of microwells is constructed with the agglutination principle of nanoprobes. Functionalized nanomaterials are also synthesized by bio-templates with high biocompatibility as bio-probe labels. The resulting analysis methodologies are simple, rapid, sensitive, cost-effective and high-throughput, enabling for one-time analysis of multiple tumor biomarkers for POC monitoring of micrometastasis. They might circumvent the analysis "bottlenecks" of classical detection means, promising new detection tools for the early warning, and POC monitoring or bedside self-checking of tumor blood-way micrometastasis.

肿瘤血行微转移是肿瘤细胞侵入血循环、常规检验技术难以检出的肿瘤转移最初阶段。其准确预警与监控对及时控制病灶、提高肿瘤的治愈率和患者放化疗与术后的存活率意义重大。项目一方面通过构造微孔筛并结合阴阳性生物磁珠,高效捕获与识别复杂血样中低丰度循环肿瘤细胞（CTC）,进而将界面磁分离法和等温信号扩增手段相结合,一步式实现靶基因序列的实时逆转录和扩增分析，构建灵敏、特异、高效的传感检测平台，用于肿瘤微转移的临床诊断与预警; 另一方面，结合光电微孔阵列和纳米探针凝集原理，并利用生物模板制得生物相容性纳米信号探针，发展简便、敏感、高通量的微孔阵列分析技术，一次性联合检获血样中多种肿瘤标志物(如特异蛋白和靶基因)，实现术后和放化疗过程中肿瘤微转移的即时监测。由此优选的传感预警平台和即时监测技术，可望突破当前相关检验技术某些瓶颈,为肿瘤血行微转移的临床诊断和现场监控及患者自查提供新的检测工具。

近年来，肿瘤、糖尿病、帕金森病、重金属中毒症等疾患发生率呈现逐年上升趋势，发展相关标志物的高效检测技术成为当务之急，特别是，肿瘤血行微转移的准确预警监控，对提高肿瘤的治愈率和患者术后的存活率意义重大。项目按照既定的研究计划，一方面，通过制备一些高性能磁性载体和纳米信号探针，并结合核酸工具酶（如核酸外切酶）选择性催化与信号倍增手段，研发了一些高效的传感检测平台，用以检测血样或尿液中低丰度的CTC细胞和microRNA等肿瘤标志物，借以满足肿瘤血行微转移的临床诊断与预警的需求；另一方面，针对肿瘤、糖尿病、重金属中毒症等重大疾患的现场监控与自查的需要，通过构建超疏水性界面并引入功能酶与纳米氧化锌等催化材料，并结合制备尺寸可控的贵金属纳米簇（如纳米金簇、纳米银簇）、功能碳材料（如石墨烯、碳纳米角）、有机功能探针（如SO2、ClO−、pH敏感探针）、和载体材料（如介孔三聚氰胺和水溶胶），发展了一批简便、特异、敏感的微孔阵列即时监测技术，用以高通量或一次性检获血/尿样中多种疾病标志物(如CTC、hela细胞、MicroRNA、血红蛋白、血糖、重金属离子、硫化氢、SO2、ClO−、有机磷)，并在有机光电探针材料和功能载体的绿色合成方法学方面，取得了一系列具有广泛影响的技术成果；由此优选的传感预警平台和即时监测技术，将为肿瘤血行微转移的临床诊断以及糖尿病等其它疾患的现场监控与自查提供可靠的检测新工具。项目发表了标注本基金资助的SCI收录论文85篇（SCI 一区27篇，二区51篇, IF6.0以上19篇，高被引论文10篇），包括Green Chem.（4篇）、 Biomaterials、 Anal. Chem.（5篇）、Chem. Commun. （5篇）、Nanoscale（3篇）、 J. Org. Chem. （8篇）、Org. Chem. Front. （5篇）、J. Mater. Chem. B（4篇）、Adv. Synth. Catal.、Biosens. Bioelectron.等国际核心期刊；首位申请国家发明专利37项（已授权6项），并有多项技术成果实现转化；组织筹办了”泰山学术论坛国际学术会议；参加国内外学术交流会议20余次；培养博、硕士研究生33名和青年骨干教师10余名；圆满地完成了研究计划的各项研究目标和任务。