Development of fully-human anti-CD30 chimeric antigen receptors

我们设计了4种不同的单链抗体分子作为抗CD30CARS的抗原识别结构域，并与这些不同的单链抗体进行了体外比较。我们选择了最佳的单链抗体，并在不同的汽车设计体外和小鼠肿瘤模型中进行了测试。我们已经确定了一种最优的scFv和汽车设计，以供进一步开发。这项工作导致了两种在小鼠身上都非常有效的汽车。我们已经选择了其中一辆车进行I期临床试验。同时完成的工作导致了编码这种汽车的临床级慢病毒载体的产生。我们已经完成了用于全人抗CD30汽车临床试验的临床方案的工作，该方案已经得到NCI机构审查委员会的批准。我们已经完成了改进抗CD30 CAR设计和T细胞培养方法的临床前工作，重点是比较不同的铰链和跨膜结构域，以及比较CD28和4-1BB共刺激结构域。表达抗CD30 CAR的T细胞的临床试验已经开始，我们已经治疗了20名患者。不幸的是，这些患者的客观有效时间最长只有3个月。由于这些短时间的反应，我们已经开发出允许一个T细胞同时针对CD30和CD19的遗传结构，目标是能够同时消除霍奇金芦德·斯特恩伯格细胞和作为芦德·斯特恩伯格细胞起源的B细胞。

We designed 4 different single chain Fv molecules as antigen-recognition domains for anti-CD30 CARs and compared CARs with these different scFvs in vitro. We selected the optimal scFv, and tested it in different CAR designs in vitro and in mouse tumor models. We have identified an optimal scFv and CAR design for further development. This work led to 2 CARs that are both highly effective in mice. We have selected one of these CARs for a phase I clinical trial. Simultaneous work has been completed that has led to generation of a clinical-grade lentiviral vector encoding this CAR. We have completed work on a clinical protocol for a clinical trial of the fully-human anti-CD30 CAR, and this protocol has been approved by the NCI Institutional Review Board. We have completed preclinical work to improve anti-CD30 CAR design and T-cell culture methods with an emphasis on comparing different hinge and transmembrane domains as well as comparing CD28 versus 4-1BB costimulatory domains . A clinical trial of T cells expressing an anti-CD30 CAR has opened, and we have treated 20 patients on this trial. Unfortunately, the longest objective duration of response in these patients has been only 3 months. Because of these short durations of response, we have developed genetic constructs that allow one T cell to simultaneously target both CD30 and CD19 with a goal of being able to eliminate both the Hodgkin Reed Sternberg cells and the B cells that are the origin of Reed Sternberg cells.