New Chimeric Antigen Receptors for Treating Hematologic Malignancies

在过去的一年中，该项目包括测试可能影响嵌合抗原受体（汽车）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γ逆转录病毒载体。 用慢病毒载体转导的T细胞具有较短的CAR表达持续时间;因此，我们决定用γ逆转录病毒载体进行未来的工作。 嵌合抗原受体由几种成分组成，通常来源于单克隆抗体的抗原识别部分，将抗原识别部分连接到跨膜部分的铰链区，共刺激结构域如4-1BB和CD 28，以及T细胞活化结构域如CD 3-ζ。 在过去的6个月里，我们已经建造了10辆新的汽车，以测试汽车的各种组件。 通过使用γ逆转录病毒载体用各种汽车转导T细胞，并进行体外测定。 其目的是找到赋予T细胞杀死癌细胞和增殖能力而不产生大量潜在毒性炎症细胞因子的汽车。我们已经发现，改变铰链区、共刺激结构域或T细胞活化结构域都会导致CAR功能的显著差异。 在广泛的体外测试之后，我们将在明年内在小鼠模型中测试有希望的汽车。这项工作还处于早期阶段，但希望它能在未来2-3年内改进用于临床测试的汽车。

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.