CD5-targeted LNP-mRNA generates CD19 CAR-T cells in the body to achieve lupus remission without clearing lymph nodes
关键词:
CAR-T cells Lipid nanoparticles mRNA In vivo generation CD19 Systemic lupus erythematosus Lymphodepletion-free CD5 targeting摘要
Conventional methods that rely on viral vectors, ex vivo cell manufacturing, and lymphocyte-depleting preconditioning significantly limit the use of chimeric antigen receptor T-cell (CAR-T) therapy, despite its potential in treating systemic lupus erythematosus (SLE). This study aimed to design an in vivo method for generating CD19 CAR-T cells utilising lipid nanoparticle (LNP)-mRNA and validate the feasibility of disease remission in lupus models without lymphodepletion. We created a CD5/LNP-CAR19 nanoformulation that is precisely delivered to T cells by encasing the mRNA encoding a second-generation CAR that targets human CD19 in LNPs coupled with CD5 antibodies using microfluidic technology.A single intravenous injection was given to MRL/lpr lupus mice as a model, and T-cell transfection efficiency, CAR expression dynamics, B-cell depletion, autoantibody levels, proteinuria, kidney pathology improvements, and safety indicators such as cytokine release were systematically evaluated using flow cytometry, in vivo imaging, ELISA, and histopathology. In peripheral T cells, CD5/LNP-CAR19 effectively and selectively produced temporary CAR expression, which lasted more than five days and reached 32.4% ± 5.1% CAR⁺ T cells 24 hours after injection. Deep and long-lasting B-cell depletion was caused by a single dose; over 95% of B cells in peripheral blood and lymphoid tissues were destroyed, and the effects persisted for more than eight weeks. Urine protein/creatinine ratios recovered to normal, kidney immune complex deposits and pathology scores were drastically reversed, and anti-dsDNA antibody titers dramatically decreased in the therapy group. In the medicated group, survival rose from 40% in the untreated mice to 90%. Throughout, no lymphocyte depletion was carried out, and there was no discernible cytokine release syndrome, neurotoxicity, or weight loss. In a lupus model without lymphodepletion, this study first demonstrated that CD5-targeted LNP-mRNA may instruct T cells in vivo to temporarily produce enough functional CD19 CAR-T cells, allowing for safe long-term remission. With substantial clinical translational potential, this "in vivo, ready-to-go" CAR-T approach provides a very accessible and safe new therapy option for B-cell-mediated autoimmune disorders.
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