基于数字化器官芯片的肝癌3D体外模型构建及其在抗肿瘤免疫治疗中的研究

Liver cancer is a severe clinical problem, and there is lack of effective therapeutic agents to improve treatment outcomes, which necessitates tremendous efforts to develop new anticancer drugs. However, conventional drug screening strategies based on 2D cell culture cannot recapitulate the key features of 3D in vivo tumor environment. As such, anticancer agents obtained through this conventional 2D platform often fail in subsequent animal studies and clinical trials. To enhance drug development, we propose to develop a digital liver-on-a-chip platform that mimics 3D in vivo tumor microenvironment in an array format under physiological flow conditions, and allows for more accurate assessment of cellular responses to anticancer agents. This digital liver-on-a-chip platform will consist of an array of 10,000 3D cell-laden hydrogel spheres of 200 µm in diameter, and the cell responses will be measured simultaneously, overcoming the drawback of bulky analysis in current organ-on-a-chip platforms. We will validate this novel 3D digital drug-screening platform using NK cell-derived exosomes. The successful development of 3D digital organ-on-a-chip platform will provide a versatile and robust toolbox to assist in screening of anticancer agents for liver cancer and other cancers.

肝癌是一个严重的临床问题，并且缺乏有效的治疗药物。然而由于2D药物筛选平台不能有效模拟体内3D肿瘤微环境，往往导致筛选出的抗癌药物无法在后续的动物研究和临床试验中产生疗效。为了提高药物的开发水平，本课题拟建立一种新型基于微阵列技术的数字化肝癌3D器官芯片，通过模拟体内3D肿瘤微环境，在动态灌注的生理条件下研究肿瘤细胞对抗癌药物/制剂的反应规律。该数字化器官芯片含有10,000个接种有肝癌细胞的水凝胶3D微球，可同步测试微球内细胞对抗癌药物的应答反应，从而能有效地克服现有器官芯片平台整体性分析，以及重复性差的缺点。该3D数字化药物筛选平台将测试NK细胞来源的外泌体对肝癌细胞的杀伤作用，并初步探索NK细胞外泌体杀伤肝癌细胞的作用机制。该3D数字化器官芯片平台的成功研发，将为肝癌和其他癌症的抗癌药物的筛选提供一个高通量和高性能的精准分析工具，从而推动器官芯片技术在精准医学及个性化医疗中的应用。

本项目利用微流控技术，通过模拟体内3D肿瘤细胞生长微环境，开发了一系列器官芯片，在动态灌注条件下探究药物对肿瘤的杀伤作用，并探索自然杀伤细胞外泌体对肿瘤细胞的杀伤作用。主要的研究内容包括制备脱细胞器官支架/GelMA新型复合水凝胶；开发多种3D器官芯片；利用器官芯片评估药物、自然杀伤细胞及其外泌体对肿瘤的杀伤作用。本项目中数字化器官芯片的开发为肝癌和其他癌症的肿瘤药物筛选提供了高通量和高精度的精准分析工具，促进了器官芯片技术在精准医学及个性化医疗中的应用。