Coradin Tiziana, Keating Amy L, Barnard Alun R, Whilding Lynsey, Pombal Diana, Hannoun Zara, Lewis Jack, Devarajan Gayathri, Iqball Sharifah, Burton Emma, Ferluga Sara, Jones Daniel M, Alberts Ben M, Wright Jordan, Farley Daniel C, O'Connor Deirdre M, Rao Ravi M, Mitrophanous Kyriacos A, Lad Yatish, Nimmo Rachael
Oxford Biomedica (UK) Ltd., Oxford OX4 6LT, UK; iosBio Ltd., Haywards Heath RH16 1DB, UK.
Oxford Biomedica (UK) Ltd., Oxford OX4 6LT, UK; Department of Oncology, University of Oxford, Oxford OX3 7DQ, UK.
Mol Ther. 2025 Jul 16. doi: 10.1016/j.ymthe.2025.07.006.
Chimeric antigen receptor (CAR) T cell therapy has proved remarkably successful for the treatment of hematological malignancies. However, the bespoke manufacturing of autologous CAR T cells is complex and expensive. The development of methods for in vivo engineering of T cells will enable generation of CAR T cells directly within the patient, bypassing the need for ex vivo manufacturing and thereby enabling greater access for patients. Here, we describe development of an improved retargeted Nipah envelope system paired with a fourth-generation lentiviral vector capable of specifically targeting T cells with increased efficiency, which generates high levels of functional CAR T cells in vivo. The retargeted vectors exhibited greater specificity to T cells compared to the VSV-G pseudotyped vector. Vectors targeted to either CD3 or CD8 similarly generated high levels of CAR T cells, which rapidly eradicated B cells, suggesting that T cell receptor (TCR) engagement is not required for lentiviral vectors to efficiently transduce T cells in vivo. Furthermore, the fourth-generation lentiviral vector platform (referred to as the TetraVecta system) employs the TRiP system to prevent incorporation of CAR protein into the vector particles, minimizing the risk of inadvertent transduction of tumor cells.
嵌合抗原受体（CAR）T细胞疗法已被证明在治疗血液系统恶性肿瘤方面非常成功。然而，自体CAR T细胞的定制生产复杂且昂贵。T细胞体内工程方法的开发将能够直接在患者体内生成CAR T细胞，绕过体外生产的需求，从而使更多患者能够接受治疗。在此，我们描述了一种改进的靶向尼帕包膜系统与第四代慢病毒载体的开发，该系统能够以更高的效率特异性靶向T细胞，在体内产生高水平的功能性CAR T细胞。与水泡性口炎病毒糖蛋白（VSV-G）假型载体相比，靶向的载体对T细胞表现出更高的特异性。靶向CD3或CD8的载体同样产生了高水平的CAR T细胞，这些细胞迅速清除了B细胞，这表明慢病毒载体在体内有效转导T细胞并不需要T细胞受体（TCR）的参与。此外，第四代慢病毒载体平台（称为TetraVecta系统）采用TRiP系统来防止CAR蛋白掺入载体颗粒，将肿瘤细胞意外转导的风险降至最低。
