高灵敏检测脑胶质瘤外泌体中小分子药物代谢的微流控芯片/质谱新技术

Glioma is an aggressive brain tumor (accounting for 40~50% in all of brain tumor patient cases). It is difficult to be eradiated completely by surgery, exhibiting multiple drug resistance during chemotherapeutic treatment. Therefore, there is an urgent request to develop personalized therapies for the glioma patients. This project proposes a new concept: important clues for glioma drug resistance research may be discovered by analyzing the small molecular drugs and their metabolites in the glioma exosomes. Being benefited from our previous work on biomedical applications of microfluidic devices and biomarker detection, we plan to develop an advanced platform combining microfluidics and mass spectrometry for highly sensitive detection of Temozolomide (TMZ) and its potential metabolites in glioma exosomes. In this new project, we will design a microfluidic device featured with three-dimensional micro/nanostructures in the sample loading channel to enrich the exosomes with a high efficiency. Interestingly, exosomes can also be released off the microchip for recovery in a well-controlled manner. The small molecular drugs or their metabolites can be detected by our graphite dots-assisted MALDI mass spectrometry which offers unique advantages of broad-spectrum, highly sensitive, and quantitative analysis. Therefore, the platform is a powerful tool for research of TMZ metabolism and glioma drug resistance. It can be further applied onto the exosome analysis targeted for the cerebrospinal fluid samples from glioma patients, which promises important clinical utilization by facilitating chemotherapy monitoring and development of personalized medicine.

恶性脑胶质瘤是最常见的颅内恶性肿瘤（占颅脑肿瘤的40~50%），手术较难根除，对多种化疗药物有耐药性，因此患者对个性化医疗的需求十分迫切。本项目创新性提出分析脑胶质瘤外泌体中的小分子药物及其代谢产物，可为研究脑胶质瘤耐药性提供重要线索。在微流控芯片生物医学应用和疾病标志物检测的前期工作基础上，我们拟发展微流控芯片/质谱新技术高灵敏鉴定分析脑胶质瘤外泌体中的替莫唑胺（TMZ）小分子原药及其代谢产物。设计含3D微纳结构组合的微流控芯片高效捕获富集外泌体并可释放回收，利用石墨点纳米材料辅助的MALDI质谱对外泌体中的小分子代谢物进行广谱、高灵敏、定量的分析检测，研究TMZ代谢过程的变化与脑胶质瘤的耐药性。本项目发展的技术可以进一步为脑胶质瘤患者脑脊液样本提取的外泌体开展有针对性的检测，将有力地推动化疗效果监控、便于为患者制定精准及时的个性化治疗方案，具有重要的临床应用前景。

肿瘤细胞外泌体是一类重要的循环肿瘤标志物，其囊泡结构所含的药物分子的浓度与代谢变化可为肿瘤耐药研究提供有用线索。本项目发展了微流控芯片/质谱新技术，并应用于高效、选择性富集肿瘤细胞所分泌的不同种类的囊泡结构，可以对外泌体和大尺寸囊泡分别回收利用，并对囊泡中所含的药物分子进行高灵敏鉴定分析，探讨了囊泡生成与肿瘤细胞耐药发生关联的情况。本项目发展了多种新型纳米基质辅助的MALDI质谱技术平台，对脑胶质瘤重要靶点蛋白进行高通量小分子药物筛选，成功筛选到高效抑制剂作为候选药物，并研究了该小分子化合物对脑胶质瘤代谢重编程的调控影响，以及小分子药物的药代动力学过程。本项目还开展了多种方法学的研究并应用于循环肿瘤标志物的检测，有助于促进快筛检测和疗效监控、发展个性化的肿瘤治疗医学。