基于细胞表型与基因表达谱分析的高通量微流控药物筛选技术研究

In vitro cell-based drug screening is of utmost importance to the discovery of new drugs. However, the current high-throughput drug screening platforms utilizing robotic systems are facing great challenges: The increasing compound libraries demand low-cost, ultra-high-throughput, and easy-to-use screening systems, and the mechanism-based screening strategy requires comprehensive omics analysis operated in a high-throughput manner to improve the screen efficiency. In our previous study, we have successfully developed a novel superhydrophobic microwell array platform, on which high-throughput cell culture was achieved. However, the limited image-based analysis on this system cannot fulfill the requirement of next-generation drug screening. Here we propose to develop a new superhydrophobic microwell array which combines oil and superhydrophobic porous polymer together. With the aid of drug delivery using photo-polymerized thermos-sensitive hydrogels, high-throughput cell phenotypic drug screening will be realized on this new system. In addition, we will develop a novel high-throughput sequencing sample preparation technology for digital gene expression profiling using the superhydrophobic microwell array. Thousands of samples can be prepared on the chip simultaneously for high-throughput sequencing. By linking the drug screening with DNA sequencing technology, we will establish a new precision drug screening strategy based on the comprehensive analysis of cell changes in both the phenotype and the gene expression levels.

体外细胞水平的药物筛选对于新药开发具有重要意义，然而基于机械手操作的传统高通量筛选技术面临新的挑战：不断扩大的药物分子库需要成本更低廉、通量更高、且易于操作的新型技术平台；而以药物作用机制为依据的筛选需要引入更为全面的高通量组学技术手段来提高筛选效率。申请人前期开发了基于超疏水原理的微孔阵列芯片技术，实现了芯片上的高通量细胞培养。但该系统仅以成像技术为分析手段还无法满足下一代药物筛选的需求。本课题拟开发油-聚合物复合材料与三维微结构相结合的新一代超疏水微孔阵列芯片，利用光聚合温敏水凝胶做为药物添加载体，实现基于细胞表型分析的高通量药物筛选。在此基础上，利用超疏水微孔阵列芯片开发高通量数字基因表达谱测序样本建库技术，实现上千个样品的并行制备，将药物筛选与高通量测序结合在一起，在高通量数据处理技术的支持下，建立一种新型的基于细胞表型与全基因组表达深度融合分析的高通量精准药物筛选技术平台。

体外细胞水平的药物筛选对于新药开发具有重要意义，然而基于机械手操作的传统高通量筛选技术面临新的挑战：不断扩大的药物分子库需要成本更低廉、通量更高、且易于操作的新型技术平台；而以药物作用机制为依据的筛选需要引入更为全面的高通量组学技术手段来提高筛选效率。申请人前期开发了基于超疏水原理的微孔阵列芯片技术，实现了芯片上的高通量细胞培养。但该系统仅以成像技术为分析手段还无法满足下一代药物筛选的需求。本课题开发了新一代超疏水微孔阵列芯片和3D肿瘤类器官的体外培养技术，并基于此开发了超疏水微孔阵列的细胞培养与药物处理平台，实现基于细胞表型分析的高通量药物筛选。在此基础上，利用超疏水微孔阵列芯片开发高通量数字基因表达谱测序样本建库技术，实现上千个样品的并行制备，将药物筛选与高通量测序结合在一起，在高通量数据处理技术的支持下，建立一种新型的基于细胞表型与全基因组表达深度融合分析的高通量精准药物筛选技术平台。