Winter Nora, Kolbe Carolin, Ferdos Shima, Steiner Larissa, Bartelsen Nils, Lock Dominik, Engert Fabian, Engels Boris, Assenmacher Mario, Schindler Michael, Lauer Ulrich M, Schaser Thomas, Cordes Nicole
Miltenyi Biotec B.V. & Co KG, 51429 Bergisch Gladbach, Germany.
Institute for Medical Virology and Epidemiology of Viral Diseases, University Hospital Tübingen, 72076 Tübingen, Germany.
Mol Ther Methods Clin Dev. 2025 Jul 3;33(3):101526. doi: 10.1016/j.omtm.2025.101526. eCollection 2025 Sep 11.
Targeted cell entry with lentiviral vectors (LVs) is one approach to enable efficient and specific gene delivery in the context of immunotherapies, such as chimeric antigen receptor (CAR)-T cell therapy. We have recently shown the generation of CAR-T products with varying CD4:CD8 ratios with our selective, universal Adapter-LV system. However, low viral vector titers limit the clinical and applications of our measles virus (MV)-based system. Here, we report a novel pseudotype based on canine distemper virus (CDV) for the Adapter-LV system (CDV-Ad-LV) and show improved viral vector yield while maintaining flexibility, selectivity, and transduction efficacy. As for MV, the engineered CDV envelope protein is fused to a single-chain variable fragment that binds biotin in the context of a defined chemical linker. Thereby, Adapter-LVs bind only to biotinylated adapter molecules, which in turn bind the antigen of choice. Besides extensive characterization , we evaluated transduction with CDV-Ad-LVs and showed high transduction efficiencies of CD4 and CD8 T cells following a single-dose injection into human PBMC-transplanted immunodeficient non-obese diabetic (NOD) SCID gamma (NSG) mice. Hence, CDV-Ad-LVs represent a novel and efficient system for adapter-mediated transduction, which broadens the potential of the Adapter-LV technology, going beyond CAR-T cell therapy applications.
使用慢病毒载体（LVs）进行靶向细胞导入是在免疫疗法（如嵌合抗原受体（CAR）-T细胞疗法）中实现高效和特异性基因传递的一种方法。我们最近利用选择性通用衔接子-LV系统展示了具有不同CD4:CD8比率的CAR-T产品的生成。然而，低病毒载体滴度限制了我们基于麻疹病毒（MV）的系统的临床应用。在此，我们报告了一种基于犬瘟热病毒（CDV）的新型假型用于衔接子-LV系统（CDV-Ad-LV），并展示了在保持灵活性、选择性和转导效力的同时提高了病毒载体产量。对于MV，工程化的CDV包膜蛋白与在特定化学接头背景下结合生物素的单链可变片段融合。由此，衔接子-LV仅与生物素化的衔接子分子结合，而衔接子分子又结合所选抗原。除了广泛的表征外，我们评估了CDV-Ad-LV的转导情况，并显示在单次注射到移植了人外周血单核细胞的免疫缺陷非肥胖糖尿病（NOD）重症联合免疫缺陷（SCID）γ（NSG）小鼠后，CD4和CD8 T细胞具有高转导效率。因此，CDV-Ad-LV代表了一种用于衔接子介导转导的新型高效系统，它拓宽了衔接子-LV技术的潜力，超越了CAR-T细胞疗法的应用范围。
