通过肌注将携带功能基因的质粒传递到骨骼肌细胞中，生产目的蛋白并分泌到胞外、到达靶位点行使功能，是一种易操作、有前景的新策略。该体系需整合高效基因传递材料、精确构建的携带治疗基因的高效表达质粒及合适的基因传递方法等。基于前期发现，我们参考细胞膜结构，通过计算机模拟，优化设计两亲性三嵌段聚合物分子，制备高效、安全、经济、稳定的基因传递材料。再整合骨骼肌原位基因传递/表达体系中的各种有利因素，构建有治疗潜力的表达体系。同时，用骨骼肌特异性高效表达质粒来构建多种治疗质粒，以表达共识别肿瘤细胞和T细胞的双靶向衔接抗体、协同刺激因子以及免疫检查点阻断分子。最后，分别建立鼠源和人源细胞的白血病小鼠模型，将治疗质粒传递到骨骼肌细胞中以表达目的蛋白，探索对白血病的联合免疫治疗。项目的实施，有望建立可应用的骨骼肌原位基因表达体系，获得有效、价格低廉的白血病免疫疗法，并可为更多种疾病开发便捷、低廉的治疗方法。

It is an simple and promising strategy to produce functional proteins via intramuscular delivery of plasmids carrying functional genes to skeletal muscle cells. The proteins have been accurately designed so that they can be secreted into the blood and transferred to target sites for therapeutic purpose. The system needs to integrate efficient gene delivery materials, accurately constructed efficient expression plasmids carrying therapeutic genes and appropriate gene delivery methods. Based on our previous findings, we will optimize the design of amphiphilic triblock polymers, with cell membrane as reference, by computer simulation to design efficient, safe, economical and stable gene delivery materials. By integrating various favorable factors in the in situ gene transfer/expression system for skeletal muscle, the expression system with therapeutic potential will be constructed. Meanwhile, skeletal muscle specific and efficient expression plasmids will be used to construct a variety of therapeutic plasmids to produce the bispecific antibodies targeting both tumor cells and T cells, co-stimulators and checkpoint blocking molecules. Finally, mouse models of leukemia from mouse and human cells were established, and the therapeutic plasmids were transferred to skeletal muscle cells to express the target protein, so as to explore the combined immunotherapy for leukemia. The implementation of the project is expected to establish an applicable skeletal muscle-based gene expression system, to obtain effective, low price leukemia immunotherapy method, and to develop convenient and cheap therapeutic methods for more diseases.