Lipid nanoparticles produce chimeric antigen receptor T cells with interleukin-6 knockdown in vivo

• Published in: Journal of controlled release Vol. 350; pp. 298 - 307
• Main Authors: Zhou, Jing-e, Sun, Lei, Jia, Yujie, Wang, Zhehao, Luo, Tengshuo, Tan, Jingwen, Fang, Xiaoyan, Zhu, Hongjia, Wang, Jing, Yu, Lei, Yan, Zhiqiang
• Format: Journal Article
• Language: English
• Published: Elsevier B.V 01.10.2022
• Subjects: antibodies; antigens; antineoplastic activity; CD3 antibody; Chimeric antigen receptor T cells; Cytokine release syndrome; genes; IL-6 shRNA; interleukin-6; intravenous injection; leukemia; Lipid nanoparticles; lipids; nanoparticles; plasmids; small interfering RNA; Chimeric antigen receptor T cells; Cytokine release syndrome; IL-6 shRNA; CD3 antibody; Lipid nanoparticles
• DOI: 10.1016/j.jconrel.2022.08.033
• ISSN: 0168-3659; 1873-4995

Chimeric receptor T cells (CAR-T) can effectively cure leukemia; however, there are two limitations: a complicated preparation process ex vivo and cytokine release syndrome (CRS). In this study, we constructed a lipid nanoparticle system modified by CD3 antibody on the surface, loading with the plasmid containing the combination gene of interleukin 6 short hairpin RNA (IL-6 shRNA) and CD19-CAR (AntiCD3-LNP/CAR19 + shIL6). The system targeted T cells by the mediation of CD3 antibody and stably transfected T cells to transform them into CAR-T cells with IL-6 knockdown, thus killing CD19-highly expressed leukemia tumor cells and reducing CRS caused by IL-6. In vivo experiments showed that AntiCD3-LNP/CAR19 + shIL6 could stably transfect T cells and produce CAR-T within 90 days to kill the tumor. This significantly prolonged the survival time of leukemia model mice and demonstrated the prepared LNP exhibited the same anti-tumor effect as the traditional CAR-T cells prepared ex vivo. In this study, CAR-T cells were directly produced in vivo after intravenous injection of the lipid nanoparticles, without the need of using the current complex process ex vivo. Additionally, IL-6 expression was silenced, which would be helpful to reduce the CRS and improve the safety of CAR-T therapy. This method improves the convenience of using CAR-T technology and is helpful in further promoting the clinical application of CAR-T. [Display omitted] •New technology is required for CAR-T cells to avoid its complex preparation process and control CRS.•The LNPs modified by CD3 antibody transfected T cells and produced CAR-T cells in vivo.•The CAR-T cells produced by LNPs showed an anti-tumor effect and reduced the incidence of CRS.