New Chimeric Antigen Receptors for Treating Hematologic Malignancies

在过去的一年里，这个项目包括测试可能影响嵌合抗原受体(CARS)功能的多种因素。首先，我们比较了慢病毒载体和我们目前临床试验中使用的伽马逆转录病毒载体。转导慢病毒载体的T细胞表达CAR的持续时间较短；因此，我们决定使用伽马逆转录病毒载体进行未来的工作。嵌合抗原受体由几个部分组成，抗原识别部分通常来源于单抗，连接抗原识别部分和跨膜部分的铰链区，共刺激结构域，如4-1BB和CD28，以及T细胞激活结构域，如CD3-Zeta。在过去的6个月里，我们制造了10辆新车，以测试汽车的各种部件。用不同的CARS通过伽玛逆转录病毒载体转导T细胞，并进行体外检测。其目的是找到能够赋予T细胞杀死癌细胞并在不产生大量潜在有毒炎症细胞因子的情况下进行增殖的CAR。我们发现，改变铰链区、共刺激结构域或T细胞激活结构域都会导致CAR功能的深刻差异。在广泛的体外测试之后，我们将在明年内在小鼠模型上测试有前景的汽车。这项工作还处于早期阶段，但有希望在未来2-3年内改进临床测试的CARS。

In the past year, this project has consisted of testing multiple factors that could affect the function of chimeric antigen receptors (CARs). First we compared lentiviral vector to the gammaretroviral vector that we have used in our current clinical trials. T cells transduced with the lentiviral vector had shorter duration of CAR expression; therefore, we have decided to perform future work with the gammaretroviral vector. Chimeric antigen receptors consis to several components, the antigen-recognition moiety that is usually derived from a monoclonal antibody, a hinge region that connects the antigen-recognition moiety to the transmembrane portion, costimulatory domains such as 4-1BB and CD28, and T cell activation domains such as CD3-zeta. We have constructed 10 new CARs over the past 6 months to test various components of CARs. T cells are transduced with the various CARs by using a gammaretroviral vector, and in vitro assays are carried out. The aim is to find CARs that impart T cells with the ability to kill cancer cells and proliferate without producing large amounts of potentially toxic inflamatory cytokines. We have found that changing the hinge region, costimulatory domains, or T cell activation domains all cause profound differences in CAR function. Following extensive in vitro testing, we will test promising CARs in a murine model within the next year. This work is all at early stages, but hopefully it will lead to improved CARs for clinical testing within the next 2-3 years.