Universal MUC1 Targeted Immunotherapy

描述（由申请人提供）：本提案的目的是评估表达新型T细胞受体（TCR）的慢病毒基因载体，该受体直接结合癌细胞表面的MUC 1，作为治疗成人癌的新治疗剂。TCR（用于诱导其在转导的免疫细胞中表达的慢病毒载体）和靶标本身相对于提出用于治疗癌症的过继免疫疗法的标准手段是独特的。由Olivera Finn博士鉴定的MUC 1特异性TCR识别低糖基化MUC 1上的独特抗原结构（在85%的所有癌症中发现，但在正常细胞中没有发现;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，其将表达含有CD 28、CD 137（4-1BB）和CD 3-zeta链信号传导元件的嵌合抗原受体（CAR）。将在T细胞和NK细胞中评价这些载体。使用的载体系统将是Lentigen的专有骨架结构，包括最新的安全元件以及额外的选择标记，如tmpk“自杀”基因。靶点低糖基化MUC 1作为癌症的独特抗原特征已得到充分证实，并且是表达CAR的免疫细胞的理想靶点。在这项提案中，我们将测试中心假设，即具有显着抗肿瘤功效的慢病毒工程化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MUC 1+癌症。这些表达CAR的细胞将经历与在常规TCR-pepMHC相互作用期间发生的克隆扩增成效应和记忆群体相同的机制。此外，我们还将测试一个推论假设，即用CAR转导的NK细胞也将充当有效的抗MUC 1+效应细胞。通过我们对天然TCR、第一代CAR、第三代CAR以及激活免疫细胞用于慢病毒转导的不同方式的详细分析以及随后对效应子功能的评估，我们将创建一种确定的产品，其具有可转化为临床环境的一致的实施其用途的方法。我们将评价免疫细胞活化的标准方法（IL-2加抗CD 3）、新一代方法（抗CD 3/抗CD 28微珠）和尖端技术（基于细胞的人工抗原呈递细胞，也包括CD 137信号传导）。我们预计，该提案中产生的结果将作为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MUC 1转化研究，使其成为开展临床试验的现实的第一步。这些试验将是第一个评估非MHC限制性TCR活性的试验，这将使MUC 1+癌症患者受益。 公共卫生相关性：这项研究计划的目标是开发一种新的抗癌药物，使我们目前没有有效治疗方法的患者受益。这种疗法是基于体外免疫细胞的激活，使激活的免疫细胞表面有一个新的受体，可以识别癌细胞，然后将这些细胞引入体内。我们预计，这些经过修饰的细胞将在重新输注后消除癌细胞，并使其他治疗失败的患者受益。这种疗法如果被证明是成功的，将对美国和世界各地的癌症患者和为他们设计治疗的医疗保健提供者产生重大影响。

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.