Engineering Cytolytic Effectors to Eliminate Virus-Infected and Cancer Cells

项目摘要 这项STTR资助申请的目标是设计强大的基于NK细胞的细胞溶解效应细胞，用于 治疗应用。这些细胞将结合T细胞的选择性和特异性以及NK细胞的能力 细胞介导对广谱异常细胞的有效细胞溶解活性，包括那些 粘附性受损。为了实现这一目标，我们打算赋予NK92细胞以 具有相同特异性的完整T细胞受体(TCR)-CD3复合体或嵌合抗原受体(CAR)。在……里面 在我们之前的工作中，我们开发了“体外进化”方法来增加天然TCR的亲和力。vbl.使用 通过这种方法，我们已经开发出针对不同病毒或癌症的高亲和力TCR和CAR- 相关表位。而这两种受体都以高亲和力结合抗原，受体介导的 由于受体细胞内信号域的不同，信号会有所不同。信号传递的动力学 由这些受体的两个不同的信号域介导，预计会影响效率，并可能 NK92-TCR-CD3和NK92-CAR细胞裂解效应器对靶细胞裂解的敏感性。因为这个基因- 修改后的NK92细胞在转移给患者之前必须进行照射，它们将能够进行运动 对靶细胞具有最多5天的细胞杀伤活性。因此，靶细胞破坏的动力学成为一种 决定基于NK92的细胞溶解效应器效率的关键因素。所取得的结果将使我们能够 选择NK92效应器，它将对感染病毒和 临床试验中的癌细胞。与采用TCR或 CAR，受辐射的NK92-CAR和NK92-TCR-CD3效应器的潜在毒性将降至最低 辐照效应器的有限持久性。然而，NK92效应器可以转移到患者身上 多次产生对病毒感染或癌细胞的有效活性。一种新型的溶细胞剂 效应器将大量上市，并可用于治疗不同基因的患者 背景，因此提供了比现有的T细胞过继转移疗法显著的优势。

Project Summary The goal of this STTR grant application is to engineer powerful NK cell-based cytolytic effector cells for therapeutic applications. The cells will combine the selectivity and specificity of T cells and the ability of NK cells to mediate effective cytolytic activity against a wide spectrum of aberrant cells, including those with impaired adhesion properties. To achieve this goal we intend to endow NK92 cytolytic cells with either the entire ¿¿ T-cell receptor (TCR)-CD3 complex or a chimeric antigen receptor (CAR) with the same specificity. In our previous work we developed "in vitro evolution" approaches to increase the affinities of native TCRs. Using this approach, we have developed high-affinity TCRs and CARs that are specific for different viral- or cancer- associated epitopes. While both of these receptors bind the antigen with high affinity, receptor-mediated signaling will differ due to differences in the receptor's intracellular signaling domain. The kinetics of signaling mediated by two distinct signaling domains of these receptors is expected to affect the efficiency, and possibly the sensitivity, of target cell lysis by NK92-TCR-CD3 and NK92-CAR cytolytic effectors. Because the gene- modified NK92 cells have to be irradiated prior to their transfer to patients, they will be capable of exercising cytolytic activity against target cells for up to 5 days. Thus, the kinetics of target cell destruction becomes a critical factor determining the efficiency of NK92-based cytolytic effectors. The results obtained will enable us to choose the NK92 effector that will exercise the most efficient cytolytic activity against virus-infected and cancer cells in clinical trials. As opposed to adoptively transferred peripheral T cells with either a TCR or a CAR, the potential toxicity of the irradiated NK92-CAR and NK92-TCR-CD3 effectors will be minimized due to the limited persistence of the irradiated effectors. However, NK92 effectors could be transferred to a patient multiple times to generate effective activity against the virus-infected or cancer cells. The novel cytolytic effectors will be available in large amounts and could be utilized for treatment of patients with different genetic backgrounds, thus providing significant advantage over existing T cell adoptive transfer therapies.