Universal MUC1 Targeted Immunotherapy

描述（由申请人提供）：本申请的目的是评估表达一种新型T细胞受体（TCR）的慢病毒基因载体，该载体直接与癌细胞表面的MUC1结合，作为治疗成人癌症的新药物。TCR，用于诱导其在转导免疫细胞中表达的慢病毒载体，以及靶标本身，与提出用于治疗癌症的过继免疫疗法的标准手段相比，是独一无二的。由Olivera Finn博士发现的MUC1特异性TCR识别了低糖基化MUC1上的独特抗原结构（在85%的癌症中发现，但在正常细胞中没有，包括乳腺癌、前列腺癌、头颈癌和肺癌），而不是MHC结合的肽。这使得我们的TCR成为一种真正的通用试剂，不依赖于肿瘤的mhc限制性抗原呈递。我们将创建单链“第三代”载体，表达含有CD28、CD137 （4-1BB）和CD3-zeta链信号元件的嵌合抗原受体（CAR）。这些载体将在T细胞和NK细胞中进行评估。所使用的载体系统将是Lentigen的专有骨干结构，包括最新的安全元素以及额外的选择标记，如tmpk“自杀”基因。低糖基化MUC1是癌症的一个独特抗原标志，是表达CAR的免疫细胞的理想靶标。在本提案中，我们将验证具有显著抗肿瘤功效的慢病毒工程T细胞可以通过开发改进的“第三代”嵌合抗原受体（CAR）来创建，该受体将靶向并特异性杀死MUC1+癌症。这些表达car的细胞将经历与传统的TCR-pepMHC相互作用过程中发生的克隆扩增成效应和记忆群体的相同机制。此外，我们还将验证一个推论假设，即经CAR转导的NK细胞也将作为有效的抗muc1 +效应细胞。通过我们对原生TCR、第一代CAR、第三代CAR的详细分析，以及激活免疫细胞进行慢病毒转导的不同方式，以及随后对效应功能的评估，我们将创造出一种最终的产品，并具有可转化为临床环境的一致实施方法。我们将评估免疫细胞活化的标准方法（IL-2加抗cd3）、新一代方法（抗cd3 /抗CD28珠）和前沿技术（基于细胞的人工抗原呈递细胞，也包括CD137信号）。我们预计该提案产生的结果将作为II期SBIR研究的基础，在此期间，我们将在I期临床试验中测试表达CAR的T细胞或T细胞+NK细胞控制或消除肿瘤生长的能力。Lentigen公司的慢病毒生产能力，结合匹兹堡大学Olivera Finn博士的MUC1转译研究，使其成为开展临床试验的现实的第一步。这些试验将首次评估非mhc限制性TCR活性，这将使MUC1+癌症患者受益。

DESCRIPTION (provided by applicant): The goal of this proposal is to evaluate lentiviral gene vectors expressing a novel T cell receptor (TCR), that binds directly to MUC1 on the surface of cancer cells, as a new therapeutic agent for the treatment of adult carcinoma. The TCR, the lentiviral vector used to induce its expression in transduced immune cells, and the target itself are unique with respect to the standard means by which adoptive immunotherapy is proposed for treating cancer. The MUC1-specific TCR identified by Dr. Olivera Finn recognizes a unique antigenic structure on hyopglycosylated MUC1 (found on 85% of all carcinomas but not on normal cells; including breast, prostate, head and neck, and lung cancer) as opposed to a peptide bound by MHC. This makes our TCR a truly universal reagent that is not dependent on MHC-restricted antigen presentation by the tumor. We will create a single chain "third-generation" vector that will express a chimeric antigen receptor (CAR) containing CD28, CD137 (4-1BB) and CD3-zeta chain signaling elements. These vectors will be evaluated in both T cells and NK cells. The vector system used will be Lentigen's proprietary backbone construct that includes the latest safety elements as well as additional selection markers such as the tmpk "suicide" gene. The target, hypoglycosylated MUC1, is well-established as a unique antigenic signature of cancer and is an ideal target for immune cells expressing a CAR. In this proposal we will test the central hypothesis that lentiviral engineered T cells with significant anti-tumor efficacy can be created by developing improved "third generation" chimeric antigen receptors (CAR) that will target and specifically kill MUC1+ cancers. These CAR-expressing cells will undergo the same mechanism of clonal expansion into effector and memory populations that occurs during conventional TCR-pepMHC interactions. Moreover we will also test a corollary hypothesis that NK cells transduced with CAR will also serve as potent anti-MUC1+ effector cells. Through our detailed analysis of the native TCR, first generation CAR, third generation CAR, and different ways of activating immune cells for lentiviral transduction and subsequent evaluation of effector function, we will create a definitive product, with a consistent means of implementing its use that is translatable to the clinical setting. We will evaluate both standard means of immune cell activation (IL-2 plus anti-CD3), newer generation methods (anti-CD3/anti- CD28 beads), and cutting edges techniques (cell based artificial antigen-presenting cells that also include CD137 signaling). We anticipate that the results generated in this proposal will serve as the basis for Phase II SBIR studies, during which we will test the ability of T cells or T cells +NK cells expressing CAR to control or eliminate tumor growth in a Phase I clinical trial. The lentiviral production capability of Lentigen Corp., combined with the translational MUC1 research by Dr. Olivera Finn at the University of Pittsburgh makes this a realistic first step in carrying out clinical trials. These trials will be the first to evaluate non-MHC restricted TCR activity that will benefit patients suffering from MUC1+ cancer. PUBLIC HEALTH RELEVANCE: The goal of this research proposal is to develop a new anti-cancer agent that will benefit patients for which we currently do not have effective therapy. This therapy is based on the activation of immune cells outside the body, giving activated immune cells a new receptor on their surface that recognizes cancer cells, and then introducing these cells back into the body. We anticipate these modified cells will eliminate cancer cells upon re-infusion and benefit patients failing other therapies. This therapy, if proved successful, will have a significant impact for cancer patients and healthcare providers designing therapy for them in the United States and worldwide.