Le Thi Hoa, Sun Fan, Xiao Gutian, Qu Zhaoxia
Norris Comprehensive Cancer Center, Hastings Center for Pulmonary Research, Department of Molecular Microbiology and Immunology, University of Southern California Keck School of Medicine, Los Angeles, CA, USA.
UPMC Hillman Cancer Center, Department of Microbiology and Molecular Genetics, University of Pittsburgh School of Medicine, Pittsburgh, PA, USA.
Bio Protoc. 2025 Sep 5;15(17):e5437. doi: 10.21769/BioProtoc.5437.
This protocol describes the preparation, administration, and analysis of a nanoparticle-based therapeutic strategy (nanoPDLIM2) in combination with PD-1 immune checkpoint blockade immunotherapy and chemotherapy for the treatment of lung cancer in mouse preclinical studies. NanoPDLIM2 uses a polyethyleneimine (PEI)-based delivery system that encapsulates PDLIM2 expression plasmids for reconstituting PDLIM2 that is repressed in tumors. This approach induces tumor immunogenicity, suppresses drug resistance, and improves treatment efficacy when used in combination with carboplatin, paclitaxel, and anti-PD-1 antibodies. The protocol describes steps for mouse lung tumor induction, nanoPDLIM2 and other therapeutic reagents' preparation and administration, and subsequent analysis of tumor burden, immune response, and toxicity, providing a reproducible approach for investigators. Key features • Comprehensive workflow for preparation and delivery of nanoPDLIM2. • Combination of nanoPDLIM2 with PD-1 blockade and chemotherapeutics for superior efficacy in lung cancer treatment. • Detailed protocols for therapeutic reagents preparation, administration, tumor examination, immune analysis, health monitoring, and toxicity evaluation in a preclinical lung cancer model.
本方案描述了在小鼠临床前研究中，基于纳米颗粒的治疗策略（nanoPDLIM2）与PD-1免疫检查点阻断免疫疗法及化疗联合用于治疗肺癌的制备、给药及分析方法。NanoPDLIM2采用基于聚乙烯亚胺（PEI）的递送系统，该系统包裹着PDLIM2表达质粒，用于重建在肿瘤中被抑制的PDLIM2。这种方法可诱导肿瘤免疫原性，抑制耐药性，并在与卡铂、紫杉醇及抗PD-1抗体联合使用时提高治疗效果。该方案描述了小鼠肺肿瘤诱导、nanoPDLIM2及其他治疗试剂的制备与给药步骤，以及随后对肿瘤负荷、免疫反应和毒性的分析，为研究人员提供了一种可重复的方法。关键特性 • nanoPDLIM2制备与递送的全面工作流程。 • nanoPDLIM2与PD-1阻断及化疗药物联合使用，在肺癌治疗中具有卓越疗效。 • 在临床前肺癌模型中，治疗试剂制备、给药、肿瘤检查、免疫分析、健康监测及毒性评估的详细方案。
